1. Blood culture or sterile tubes (yellow) 2. Coagulation tube/ Na citrate tube (light blue) / black 3. Serum tube with or without clot activator or gel (red, gold, or red-gray marbled stopper) 4. Heparin tube (green/light green) 5. EDTA tubes (lavender stopper) 6. Oxalate/fluoride tubes (gray)

1. YELLOW (SPS, ACD) 2. ORANGE (THROMBIN) 3. GREEN (HEPARIN) 4. LAVENDER AND PINK (EDTA) 5. GRAY

HEMA 1 Flashcards by PASCUAL, krizzia anne lou

The average human possesses ______

5L of blood

5-6L -total blood volume

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Composition of blood

Plasma 55%
Formed elements 45%

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4 layers of centrifuged tube

FATTY
PLASMA
BUFFY
RBC’s

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Treat all specimens such as blood, body fluids and unfixed tissues to be potentially infectious

STANDARD PRECAUTION

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most effective way of breaking the chain of infection

HANDWASHING

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HANDWASHING PROCEDURE

Wet hands and wrists thoroughly under running water
Apply germicidal soap and rub hands vigorously for at least 15 seconds (rodaks)
Rinse hands in adownward flow from wrist to fingertips
Dry hands with a paper towel
Use the paper towel to turn off the faucets

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The process of destroying PATHOGENIC microorganisms in inanimate objects

DISINFECTION

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Destruction of all forms of microbial life including spores

Sterilization

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An appropriate disinfectant is a household bleach (_________), used in a _________.

sodium hypochlorite
1:10 v/v

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The process of obtaining blood from a vein

VENIPUNCTURE

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ORDER OF DRAW

Blood culture or sterile tubes (yellow)
Coagulation tube/ Na citrate tube (light blue) / black
Serum tube with or without clot activator or gel (red, gold, or red-gray marbled stopper)
Heparin tube (green/light green)
EDTA tubes (lavender stopper)
Oxalate/fluoride tubes (gray)

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Red (glass)

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: None clotting - 30-60 mins.

Specimen type/use: Serum/ chemistry, serology

Mechanism of action: N/A

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Red (plastic)

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Clot activator

Specimen type/use: Serum/chemistry, serology

Mechanism of action: silica clot activator (⬆️ surface area of plt.)

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Lavender (glass)

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: K3 EDTA in liquid form- more preferred for PBS

Specimen type/use: Whole blood hematology

Mechanism of action: Chelates/ binds calcium

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Lavender (plastic)

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: spray dried K2 EDTA (not good for PBS but more preferred for Hct and ESR)

Specimen type/use: WB/hematology

Mechanism of action: Chelates/ binds calcium

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Pink

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Spray dried K2 EDTA

Specimen type/use: Whole blood/ blood bank and molecular diagnostics.

Mechanism of action: Chelates/ binds calcium

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WHITE

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: EDTA and gel

Specimen type/use: plasma/ molecular diagnostics

Mechanism of action: chelates/binds calcium

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Light blue

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Na citrate

Specimen type/use: plasma/coagulation

Mechanism of action: Chelates/ binds calcium

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BLACK

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Na citrate

Specimen type/use: Plasma/ ESR

Mechanism of action: Chelates/ binds calcium

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LIGHT GREEN/BLACK

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Lithium heparin and gel

Specimen type/use: Plasma/ chemistry

Mechanism of action: Inhibits thrombin

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GREEN

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Sodium heparin, lithium heparin

Specimen type/use: Plasma/ chemistry

Mechanism of action: Inhibits thrombin

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ROYAL BLUE

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Sodium heparin, K2 EDTA

Specimen type/use: Plasma/chemistry/ TOXICOLOGY

Mechanism of action: Heparin inhibits thrombin, EDTA binds calcium

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GRAY

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Sodium fluoride/ potassium oxalate

Specimen type/use: Plasma/ glucose testing

Mechanism of action: Inhibits glycolysis

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Yellow

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Sodium polyanetholesulfonate

Specimen type/use: Serum(sterile/blood culture

Mechanism of action: Inhibits
complement, phagocytes and certain antibiotics

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Yellow

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Acid citrate dextrose

Specimen type/use: Plasma/blood bank/
HLA, phenotyping and paternity testing

Mechanism of action: WBC preservative

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Tan (glass)

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Sodium heparin

Specimen type/use: Plasma/ lead testing

Mechanism of action: Inhibits thrombin

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Tan (plastic)

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: K2 EDTA

Specimen type/use: Plasma/ lead testing

Mechanism of action: Chelates/ binds calcium

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Yellow/gray and orange

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Thrombin

Specimen type/use: Serum/Chemistry

Mechanism of action: Clot activator

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Red/gray and gold

Anticoagulant/additive:
Specimen type/use:
Mechanism of action:

Anticoagulant/additive: Silica clot activator, separation gel

Specimen type/use: serum/chemistry

Mechanism of action: Silica clot activator

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EDTA containing tubes

Lavender
Pink
White
Royal blue
Tan (plastic)

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Na Citrate containing tubes

Light blue
Black

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Aka SEQUESTRENE

K3 EDTA

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Aka versene

K2 EDTA

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Aka versene

Na2 EDTA

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Short Draw——> OVERANTICOAGULATION —->shrinking RBC can result to:

False ⬇️ HCT & ESR

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Preferred concentration of Na Citrate tube?

3.2%

Sample with increase hct or underfilled tube—-> ⬇️plasma=⬇️calcium

The excess citrate will neutralize the calcium from the reagent—-> prolonged test (PT, APTT)

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Blood to anticoagulant ratio using black tube

4:1

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Inhibits thrombin
Enhances anti-thrombin-III
Causes BLUE background in PBS

HEPARIN

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Inhibits thrombin
Enhances anti-thrombin-III
Causes blue background in PBS

HEPARIN

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Contains thrombin
For STAT chem
An additive

ORANGE

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Antiglycolytic
Inhibits enolase by binding magnesium
Also inhibits urease

NaF

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Substitutes NaF if both glucose and BUN are requested

Li Iodoacetate

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Preferred concentration of OXALATE

1-2 mg/mL

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Preferred concentration of CITRATE

3.2 g/dL (0.105 M)
3.8 g/dL (0.129 M)

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Preferred concentration of EDTA

1-2 mg/mL (chem)
1-5 mg/mL (Hema)

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Preferred concentration of FLUORIDE

10 mg/mL

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Preferred concentration of HEPARIN

0.2 mg/mL

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8x inversions

YELLOW (SPS, ACD)
ORANGE (THROMBIN)
GREEN (HEPARIN)
LAVENDER AND PINK (EDTA)
GRAY

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5x inversions

RED (PLASTIC)
GOLD

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3-4x inversions

LIGHT BLUE

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Tourniquet- should be applied_________above the venipuncture site and left no longer than ______.

3-4 (7.5-10 cm)
1 minute

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TOURNIQUET APPLICATION

Cross the right side of the tourniquet over the left side OR
Place tension on the tourniquet, cross one side over the other, and slip a small loop under one side of the tourniquet

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Most common means of collecting blood specimen is through the use of an evacuated tube system.

ETS

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inhibits the use of glucose by blood cells, recommended if a delay in testing is expected for glucose (E.g., NaF and lithium iodoacetate)

Antiglycolytic agent

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Prevents blood from clotting (E.g. EDTA, Potassium oxalate)

Anticoagulant

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Enhances the clotting mechanism by providing an increased surface area for platelet activation (glass or silica) and a clotting factor such as thrombin.

Clot activator

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inert material that goes a temporary change in viscosity during the centrifugation process, provides a separation barrier between the serum or plasma and the cells

Separator gel

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Routinely used gauge needles

19-, 20- and 21-gauge needles

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Needle guage used in pediatric

23-25 gauge

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THE MOST COMMON NEEDLE SIZE FOR ADULT VENIPUNCTURE IS ______ GAUGE
WITH A LENGTH OF _______.
ADVANTAGE OF USING A 1-INCH NEEDLE
IS THAT IT PROVIDES BETTER CONTROL.

21 gauge
1 INCH(exact) or 1.0-1.5 inches (in range).

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Most common skin cleanser

70% isoprophyl alcohol

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Used for sample collection for blood measurements.

Benzalkonium chloride or nonalcoholic anti-septic

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most preferred vein
largest, closest to the surface and well anchored

MEDIAN CUBITAL

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2nd option, less anchored, however it easiest vein to palpate in obese patients.

CEPHALIC VEIN

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least anchored
close to median cutaneous nerve and brachial artery

BASILIC VEIN

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Skin puncture is performed in:

Newborns, pedia below 1y/o
Adults who are severely burned
Elderly patients

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Capillary blood values as compared to venous blood:

⬇️rbc ct.
⬇️hct.
⬇️hb.
⬇️plt. ct.
⬆️glucose and WBC ct.

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True or False:

VEINS ON THE BACK OF THE HAND AND WRIST MAY BE USED FOR VENIPUNCTURE.
VEINS ON THE UNDERSIDE OF THE WRIST CAN BE USED.
LEG ANKLE, AND FOOT VEINS MAY BE USED BUT NOT WITHOUT THE PERMISSION OF A PHYSICAIN

True
False (veins underside should never be used)
True

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capillary blood collection sites

Lateral side of the plantar surface of the heel- children
Third or fourth finger- older children or adult

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SKIN PUNCTURE PROCEDURE

BSEOS

Blood gases
Slides, unless made from EDTA microcollection tubes
EDTA microcollection tube
Other anticoagulated microcollection tubes (green or gray)
Serum microcollection tubes

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Why do we need to wipe away the first drop of blood during capillary puncture.

Prevent contamination of the specimen with tissue fluid.
Facilitate the free flow of blood

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Punctures should not be more than
________ because of the risk of bone injury or possible infection (osteomyelitis)

2 mm

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Cortisol, ACTH, Fe ______
Eosinophils ________

Cortisol, ACTH, Fe- ⬆️ in AM
Eosinophils- ⬆️ in PM/evening

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During stress, WBC ct. and acid base balance ________.

INCREASED

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During exercise, creatinine, protein, CK, AST, LD, platelet and WBC ct., HDL _______

INCREASE

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Prolong standing ________.
Matagal na nakahiga_________.

HEMOCONCENTRATION

HEMODILATION

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In smoking WBC count _______

INCREASE.

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CBC

WBC count
RBC count
Hb
Hct.
WBC Differential
RBC indices

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Measurement of _______ is one of the several tests used to diagnose and follow treatment of anemia.

Comprised of ____ heme (iron+-protoporphyrin) and ____ globin chains

HEMOGLOBIN

4 heme 4 globin

15-20g/dL- at birth
12-16 g/dL- adult women
13-18 g/dL- men

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Hemoglobin ____ in the morning

Increased

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Modified Drabkins Reagents

Potassium cyanide
Potassium ferricyanide

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Donates cyanide to hemoglobin

Potassium cyanide

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Converts Fe2+ to Fe3+

Potassium ferricyanide

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Replaced the sodium bicarbonate (in the original drabkin’s reagent, shortens the
reaction time from 15 minutes to 3 minutes (10 minutes-rodaks)

Monopotassium phosphate (KH2 PO4)

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decreases amount of turbidity resulting from abnormal proteins and improves RBC lysis

Nonionic detergent

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CYANMETHEMOGLOBIN METHOD Principle:

Potassium ferricyanide converts the hemoglobin iron from the Fe++ to Fe+++ to form methemoglobin
(Hi=hemiglobin)
methemoglobin then combines with potassium cyanide to form the stable pigment cyanmethemoglobin (HiCN)
The color intensity of this mixture is measured in a spectrophotometer at a wavelength of 540m
The optical density of the solution is proportional to the concentration of hemoglobin, all forms of hemoglobin are measured except sulfhemoglobin

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Cyanmethemoglobin method principle:

Potassium ferricyanide converts the hemoglobin iron from the Fe++ to Fe+++ to form methemoglobin
(Hi=hemiglobin)
methemoglobin then combines with potassium cyanide to form the stable pigment cyanmethemoglobin (HiCN)
The color intensity of this mixture is measured in a spectrophotometer at a wavelength of 540m
The optical density of the solution is proportional to the concentration of hemoglobin, all forms of hemoglobin are measured except SULFHEMOGLOBIN.

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Sources of error
High WBC count: >20 x 10^9/L
High plt. Count: >700 x 10^9/L

Result:
Remedy:

Result: turbidity and false high results
Remedy: centrifuge the mixture and use the supernatant

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Hemoglobin S and C

Result:
Remedy:

Result: turbidity and falsely high results

Remedy: dilute mixture 1:2 with water then multiply results by 2

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Lipemic blood

Result:
Remedy:

Result: turbidy and falsely high results

Remedy: add 0. 01 ml of patient’s plasma to 5.0 mL of HiCn reagent and use this mixture as a the blank

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•Measures functional hemoglobin only.
•Based on the fact that 1g of Hb carries ______ ml of oxygen

Blood Oxygen Capacity (Gasometric/ Van Slyke Method)

1g of Hb= 1.34 mL of oxygen
= 1.39 mL of oxygen (Rodriguez)

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Blood iron content
100g of Hb=_______ of Fe2+

100g of Hb= 0.347 g

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•Used for blood donor screening

•The density of the drop of blood is ________ to the amount of Hb
If the hemoglobin is _________ , the drop of blood will sink within _______ and the donor is accepted.

The specific gravity of the copper sulfate solution is ______.

The drop of blood should be added from a height of about ______.

Copper Sulfate Method (Gravimetric method)

Directly proportional

≥12.5 g/dL

15 minutes

1.053

1 cm

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Colorimetric method
•Acid hematin (Sahli’s Method) obsolete
Reagent: _______
> A comparator block is used to compare the _________ color of the resulting solution

•Alkali hematin
Reagent: ________
HbF is ________ and therefore it can’t be used for Hb determination of newborns

ACID HEMATIN
Reagent: 0.1 N HCl
Brownish-yellow

ALKALI HEMATIN
Reagent: 0.1 N NaOH
Alkali resistant

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Formed by combination of hemoglobin with CARBON MONOXIDE
Unable to transport oxygen
Affinity for carbon monoxide is ______ greater than for oxygen
The formation is ______
Has a brilliant ______ color
Peak absorbance at ________

CARBOXYHEMOGLOBIN
200x
Reversible
Cherry red
576 nm

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Ferrous ion has been oxidized to ferric state
Incapable of transporting oxygen molecule
Reversible
Most cases are acquired primarily due to exposure to certain drugs and chemicals quinones, chlorates)
Can cause _______ discoloration of the blood.
Peak absorbance at _____

METHEMOGLOBIN
5. CHOCOLATE BROWN
6. 630 nm

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Not normally found in the blood
Formation is ________, it remains for life in the red blood cell of the carrier.
It is thought to be formed by the action _______and ______.
Can combine with carbon monoxide to form carboxysulfhemoglobin
Causes a __________ discoloration of the blood
Peak absorbance at _______nm

SULFHEMOGLOBIN
2. Irreversible
3. Sulfonamides and aromatic amines
5. Mauve-lavender or green (rodaks)
6. 618 nm

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• ________\is the volume of packed RBs that occupies a given volume of whole blood
• It is either reported as a percentage (36% or in liters per liter (.36L/L)

Hematocrit

45-60%- At birth
36-48%- Females
40-55%- Males

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MICROHEMATOCRIT METHOD

Microhematocrit tube- _______ long with an internal bore of ______, can hold ______ml of blood

75 mm long
1.2 mm internal bore
0.05 ml of blood capacity

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Contains heparin (anticoagulated tube), to be used for samples that are non-anticoagulated.

RED BAND

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Plain tubes (non-anticoagulated tube), to be used for samples that are anticoagulated.

Blue band

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Clay like sealing compound

4-6 mm

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Microhematocrit centrifuge capable of _______ RCF for _______ minutes.

10k-15k RCF
5 minutes

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Microhematocrit specimen:

________ whole blood is preferred
K3-EDTA causes a _____ decrease in the hematocrit due to shrinkage of the RBCs

K2-EDTA

2-3%

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incomplete sealing (less than 4-6 mm), leads to ________

falsely low results

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Inadequate centrifugation (shorter than 5 minutes), leads to a __________.

falsely increased result

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Allowing the tube to stand longer than several minutes leads to ______.

falsely increased result

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Overanticoagulation can cause hematocrit ______

Falsely low results

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After centrifugation, a small amount of plasma remains in the PC, and is usually expressed as a percentage of the RBC column
Encounter only in manual method

TRAPPED PLASMA

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Increased amount of trapped plasma is found in:

Macrocytic anemia
Spherocytosis
Thlassemia
Hypochromic anemia
Sickle cell anemia

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Rule of three applies to specimen that have _______ erythrocytes

Normocytic, normochromic

Rule of Three:
Rbc x 3= Hemoglobin
Hb x 3= Hct +/- 3

1 hct= 0.34 g Hb per 100 mL of WB
1 hct = 107,000 RBC’s/ cumm

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RBC Ct. normal values:

• 5.0-6.5 X 1012/L- N.V. for newborns
• 3.6-5.6 X 1012/L- N.V for females
• 4.2-6.0 X 1012/L- N.V. for males

Highest in AM
Lowest in the PM

⬆️ in PV and in patients who live in places at a high altitude

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Thoma red count pipet marks and there dilution:

0.5 (1:200)
101 (1:100)

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RBC count diluting fluid

1.Gower
2. Eagle
3. NSS
4. TOISON
5. STRONG
6. BETHEL
7. HAYEM
8. DACIE

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Hemocytometer

Consists of two identically ruled platforms
The space between the top of the platform and the cover glass over it is ____.
Each of the two platforms are composed of ______ which measure ___ wide and ____ long;
Therefore, the entire ruled area is ______
The volume of one entire platform is _______ (3mm x 3mm x 0. 1 mm)
The volume of one large square is ______
The large middle square containing 25 smaller squares is used for RBC count
The volume of each 25 smaller squares is _______ for a total volume of _______ five small squares
The four large corner squares, each of which is divided into ____ smaller squares, and are used for counting
WBC

0.1 mm
9 large squares; 1mm wide and 1 mm long
9 mm2
0.9 ul
0.1 ul
0.004 ul; 0.02
16

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RBC count formula:

RBC/L= #cells in 5 squares x VCF x Dilution factor

Ex. Blood was aspirated up to the 5 mark of the RBC thoma pipet and diluted up to the 101 mark. 400 cells were counted on the first platform using 5 BC squares, and 415 cells were counted on the second platform using the same technique in the first chamber. Compute for the RBC count.

Given:
Ct. 1= 400
Ct. 2= 415
Dilution 200
5 rbc squares used on each count

VCF= 1/#squares (vol.)
———> get the average 400+415= 407.5

407.5/5(0.004)= 4,075,000/ul x 10^6
=4.075x10^12/L (if average)

815 x 200/10 (0.004) = 163,000/ 0.04
= 4.075 x 10^12/L

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WBC Count NV:

4.0-11.0 x 10°/L- N.V. for adults
10.0-30.0 x 109/L- N. V. for newborns
6.0-17.0 x 10°/L- N.V. at 1 year of age

Higher in the afternoon
Lower in ptx. who are exposed to radiation or patients undergoing certain drug therapy

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In WBC counting, allow the dilution to sit for ______ to ensure that the red blood cells (RBCs) have lysed.
Leukocyte counts should be performed within _____ of dilution

10 minutes
3 hours

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WBC thoma pipet marks.
Dilution
White count diluting fluids

WBC thoma pipet marks
-0.5
-11

Dilution (1:20)

White count diluting fluids
2% acetic acid
1% HCl
Turk’s diluting fluid

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WBC count formula
Corrected WBC Count

WBC count= # of WBCs counted x VCF x Dilution Factor

CWC: Performed when 5 or more nucleated RBCs are present in the PBS
FORMULA:Corrected WBC ct.= (Uncorrected WBC count x 100%) /
(100 + # of nRBCs per 100 WBC)

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WBC estimation DILUTION FACTOR

2000

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Dilution for Normal WBC count

1:10 or 1:20

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If WBC is >30 x109/L

1:101 (0.02 mL blood + 2.0 mL diluent)
1:100 (aspirate blood up to 1 mark and dilute up to 101 mark in the RBC Thoma pipet)

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If WBC is 100-300 x10^9/L

1: 201 (0.02mL blood + 4 mL diluent)
1:200 (aspirate blood in RBC thoma pipet up to the 0.5 mark and dilute up to
101)

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If WBC is below 3.0 x10^9/L

1: 11 (0.02 mL blood + 0.2 mL diluent)
1:10 (aspirate blood up to 1 mark and dilute up to 11 mark in WBC Thoma pipette)

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WBC

Diluent:
Dilution:
Objective:
Area:

Diluent:
1% ammonium oxalate, 3% acetic acid, 1% HCL

Dilution:
1: 20 or
1:100

Objective: 10x

Area: 4mm2 or 9mm2

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RBC

Diluent:
Dilution:
Objective:
Area:

Diluent: ISOTONIC SALINE
Dilution: 1:100
Objective: 40x
Area: 0.2 mm^2

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Platelets

Diluent:
Dilution:
Objective:
Area:

Diluent: 1% ammonium oxalate
Dilution: 1:100
Objective: 40x, phase
Area: 1 mm2

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Size: 10-15 um

Nucleus: Segmented into 2-5 lobes (2-4 lobes)

Cytoplasm: Stains light pink, grainy appearance.

Other names: Seg, polymorphonuclear neutrophil, poly, PMN

NEUTROPHIL

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Size: 10-15 um

Nucleus: elongated, curved or sausage shaped.

Cytoplasm: Identical to segmented neutrophil.

Other name: Nonsegmented neutrophil, neutrophil staff or stab

BAND

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Size: 12-17 um

Nucleus: Dark purple, band shaped or segmented with only two lobes

Cytoplasm: Contains large, spherical granules that stain orange-pink

CONTAINS MAJOR BASIC PROTEIN

Other name: Acidophil (affinity for the acidic dye or eosin)

EOSINOPHIL

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Size: 10-14 um

Nucleus: Light to purple staining, usually difficult to see due to overlying granules

Cytoplasm: Densely stained, dark violet granules

BASOPHIL

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All of the following shift to the left, except:
A. Myelocyte
B. Metamyelocyte
C. Band (if >6)
D. Segmenters

D. Segmenters

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Largest cell in the PBS

Size: 12-20 um

Nucleus: Round, horseshoe -shaped or lobulated, usually folded or with convolutions

Cytoplasm: Abundant cytoplasm with gray-blue containing indistinct granules giving it a ground glass appearance

MONOCYTE

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Size: Small (6-8 um), medium to large (8-12 um).

Nucleus: Deep purple, compact, densely packed clumps, may be round oval, or indented

Cytoplasm: Stains pale to bright sky blue, may contain a few prominent reddish (azurophilic) granules (ROBIN’s EGG BLUE)

LYMPHOCYTE

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If the differential count shows the presence of IMMATURE granulocytes, this is termed __________ and may be found in disorders such as leukemias and bacterial infections.

Shift to the LEFT

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A shift to the right refers to an ________

increased number of hyper segmented neutrophils

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Granulocytes

Neutrophil
Eosinophil
Basophil

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Non-granulocytes

Monocytes
Lymphocytes

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Polymorphonuclear

N
E
B

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Mononuclear

M
L

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Phagocytes

N
E
B
M

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Immunocyte

Lymphocyte

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NEUTROPHILIA, except:
a. Appendicitis
b. Allergies
c. Myelogenous leukemia
d. Bacterial infection

B. Allergies

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Neutropenia

Decreased neutrophil production
——-Inherited stem cell disorder
——-Acquired stem cell disorder (benzene poisoning)

Increased neutrophil destruction
——-Certain bacteria
——-Viral

Immune reactions
——-Autoimmune
——-Isoimmune
——-Drug-induced

Sequestration

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EOSINOPHILIA, except:
a. Parasitic infections
b. Scarlet fever
c. Allergies
d. Immunodificiency

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Eosinopenia, except:
a. Brucellosis
b. Decreased production
c. Acute bacterial infection
d. ACTH administration

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Brucellosis
Tuberculosis
Subacute Bacterial Endocarditis
Typhoid
Rickettsial infections
Hodgkin’s disease
Gaucher disease

MONOCYTOSIS

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Glucocorticoids
Overwhelming infections that also cause neutropenia

MONOCYTOPENIA

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Viral infections
Whooping cough
Infectious mononucleosis IM
Lymphocytic leukemia

LYMPHOCYTOSIS

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Stress
Hyperthyroidism
Increased glucocorticoid levels

BASOPENIA

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Long-term drug therapy
Immunodeficiency

LYMPHOCYTOPENIA

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Immediate hypersensitivity reactions
Hypothyroidism

BASOPHILIA

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Indicates the average volume of RBCs in femtoliters (fL)

MCV

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MCV FORMULA
NV

hct/rbc ct. x 10
80-100 fL

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An expression of the average concentration of hemoglobin
in red blood cells

MCHC

Hb/hct x 100

NV: 32-36 g/dL

> 36 g/dL= spherocytic/hyperchromic

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Indicates the average weight of hemoglobin in the red blood cells
Not considered in the classification of anemias

MCH

Hb/rbc ct. x 10

RR: 28-32 pg

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Determined from the RBC histogram; coefficient of variation of the MCV

RDW

RR: 11.5-14.5%

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⬆️ RDW

post-transfusion, post-treatment (Fe supplements, vit. B12, or folic acid therapy), idiopathic sideroblastic anemia, presence of two deficiencies (iron and folic acid deficiency)

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MICROCYTIC, HYPOCHROMIC

ATIS
Anemia of chronic inflammation
Thalassemia
IDA
Sideroblastic anemia

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NORMOCYTIC, NORMOCHROMIC

AHA
1. Aplastic anemia
2. Hemolytic anemia
3. Acute blood loss anemia

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MACROCYTIC, NORMOCHROMIC

LMM
1. Liver disease
2. Myelodysplasias
3. Megaloblastic anemia

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Most convenient and most commonly used type of film in PBS

The size of the blood drop must be _____

The angle must be _____

Manual Wedge Technique

2-3 mm

30-45 ° or 25-40 °

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Qualities of a Properly Made Wedge Smear

The film is 2/3 to ¾ of the slide
The film is finger shaped not bullet shaped, Without holes or irregularities Rainbow colors.

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FACTORS THAT AFFECT THE THICKNESS OF A WEDGE SMEAR

Pressure
Angle
Size of blood drop
Speed

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Thick smear

P ⬇️
A ⬆️
S ⬆️
S ⬆️

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Thin smear

P ⬆️
A ⬇️
S ⬇️
S ⬇️

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Romanowsky stain

Pure wright stain
Wright giemsa stain

Polychrome stain (methylene blue, eosin)

Methylene blue (basic)- stains RNA
EOSIN (acid)- stains cytoplasm (hgb, eosinophilic granules)

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EXCESSIVELY BLUE STAIN

Thick films
Prolonged staining time
Inadequate washing
Too high alkalinity of buffer

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EXCESSIVELY PINK STAIN

Insufficient staining time
Prolonged washing time
Mounting the coverslip before the slide is dry
High acidity of the stain

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Method of PBS examination
1. The slide is moved from side to side ________

is moved tail towards the head of the smear _______
Uses a pattern of consecutive fields beginning near the tail on a horizontal edge: count three consecutive horizontal edge fields, count two fields towards the center of the smear, count two fields horizontally, count two fields vertically to the edge. A.k.a SERPENTENE/ TRACK

Cross sectional or crenellation
Longitudinal method
Battlement method

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Important in helping to diagnose bleeding disorders.

Platelet count
NV: 150,000-450,000/ uL (150-450 x 10^9/L)

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THROMBOCYTOSIS

PICS
1. Polycythemia vera
2. Idiopathic thrombocythemia
3. Chronic myelogenous leukemia (CML)
4. Splenectomy

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THROMBOCYTOPENIA

TAAG PS
1. Thrombocytopenia purpura
2. Aplastic anemia
3. Acute leukemia
4. Gaucher’s disease
5. Pernicious
6. Splenomegaly

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MPV should be less than ____ hours

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If >4 hours old MPV will have _____ increment.

20%

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Reference method for plt. Count?

Whole blood is diluted with _______, which hemolyzes red cells.

Platelets are counted using _________microscope

EDTA can cause platelet satellitosis, this can be corrected by using _______as the anticoagulant and multiply the platelet count by _____

PHASE MICROSCOPY/ BRECHER- CRONKITE

1% ammonium oxalate

Phase contrast

Sodium citrate, 1.1

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What platelet method Uses a light microscope

Rees and Ecker diluent: (BSFD)
1. Brilliant Cresyl Blue- stain
2. Sodium Citrate- anticoagulant
3. Formaldehyde- preservative
4. Distilled H20

Platelets appear as small, ROUND, OVAL or ELONGATED particles that are highly refractile and stain a light bluish color

TONKANTIN METHOD

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Place the charged hemacytometer in a moist chamber for ________ to allow the platelets to settle.

15 minutes

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____________is the last immature erythrocyte stage

Normally spends ______ days in the bone marrow and ____ day in the peripheral blood

Contains remnants of RNA and organelles such as ribosomes a day

____________is used to assess the ERYTHROPOIETIC ACTIVITY OF THE BONE MARROW.

RETICULOCYTES

2-3 days

1 day

Retics count

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Retic count formula and normal values.

%RC=# of retics per 1000 RBC’s/ 10

NV: 0.5-1.5% (adult)
2-6% (newborn)

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⬇️ retic count

aplastic anemia
conditions in which the bone marrow is not producing red blood cells

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⬆️ retic count

hemolytic anemias
individuals with IDA receiving iron therapy
thalassemia
sideroblastic anemia
acute and chronic blood loss anemia

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Retic count procedure:

Mix _____ amounts of blood and new methylene blue stain (_____drops, or ____
mcl each) and allow to incubate at room temperature for ______minutes
Remix the preparation.
Prepare two wedge films
in an area in which cells are close together but not touching, count _______RBCs under the ______ reticulocytes are included in the total RBC count (i.e., A reticulocyte counts as both an RBC and a reticulocyte).
To improve accuracy, have another laboratorian count the other smear; values should agree within _______
Calculate the reticulocyte count

SUPRAVITAL STAINS: _____

Equal, 2-3 drops, 50 ul
3-10 minutes
1000 RBCs, OIO
20%

Crystal violet
New methylene blue
Brilliant cresyl blue

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Spleen- saquesters

Sequest 20-30% of platelets

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Retic count sources of error: 3 H

Heinz body
HbH
Howell jolly

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actual number of reticulocytes in 1L of whole blood

ABSOLUTE RETICULOCYTE COUNT
ARC= (%) reticulocytes x RBC count (x10^12) ÷ 100

Reference range: 25 x 10^9/L up to 75 x 10^9/L

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CORRECTED RETICULOCYTE COUNT
In specimens with low hematocrit, the percentage of reticulocytes may be ________ because whole blood contains fewer RBCs

A correction factor is used, with the average normal hematocrit considered to be _____

Falsely elevated

45%

CRC= (%) reticulocytes x patient hct ÷ 45%

Reference range:
Hct.= 35% (CRC of 2-3%)
Hct= <25%= the count should increase to 3-5% to compensate for anemia

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Reticulocytes that are released from the bone marrow prematurely are called shift reticulocytes

Cells shifted to the peripheral blood prematurely stay longer as reticulocytes and contribute to reticulocyte count
for more than 1 day

The reticulocyte count is falsely increased because the count-no longer represents the cells maturing in just 1 day

RPI= retics (%) x [hct ÷ 45] /maturation time

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Patient hct. Correction factor?

40-45
35-39
25-34
15-24
<15

1
1.5
2
2.5
3

RPI > 3= adequate bone marrow response

RPI < 2= Inadequate bone marrow response

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A nonspeciic measurement used to detect and monitor an inflammatory response
The settling of RBCs at the bottom of a tube upon standing undisturbed for 1 hour

ERYTHROCYTE SEDIMENTATION RATE

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Rouleaux formation
Agglutination
Macrocytes
Severe anemia

INCREASED ESR

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Sickle cells and spherocytes
Anisocytosis
Poikilocytosis

DECREASED ESR

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The single most important factor in determining ESR

PLASMA COMPOSITION

ESR = RBC MASS
ESR ∝ PLASMA VISCOSITY

ex. Increased albumin = decreased ESR

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A tilt of 3° can cause errors up to ______

30%

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lower temperatures form air rushing out on opening the refrigerator or freezer

If a refrigerated blood is used for ESR the result will be?

Falsely LOW ESR

INCREASED

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vibrations from opening and closing the refrigerator doors

Falsely INCREASED ESR

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heat released from the refrigerator motor

Falsely INCREASED ESR

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WESTERGREN
Length:
Bore:
Calibration:

NORMAL VALUES:
SPECIMEN:

Length: 30 cm (300 mm)
Bore: 2.55 mm
Calibration: 0-200

NORMAL VALUES:
0-15 mm women
0-10 mm men
0-10 mm children

SPECIMEN: Na citrate whole blood (4:1)

More sensitive for patients with high ESR

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WINTROBE

Length:
Bore:
Calibration:

NORMAL VALUES:

Length: 115 mm
Bore:3 mm
Calibration:
LEFT- used for ESR (0-100 mm)
RIGHT- used for MACROHEMATOCRIT (100-0)

NORMAL VALUES:
0-20 mm/hr -women
0-9 mm/hr- men

More sensitive for patients with lower ESR

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Hypercholesterolemia
Hyperfibrinogenemia
Hypergammaglobulinemia
Hypoalbuminemia

INCREASED ESR

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Multiple myeloma
Rheumatoid arthritis

INCREASED ESR

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Acanthocytosis
Anisocytosis (marked)
Hemoglobin C
J. Microcytosis
Polycythemia
Sickle cells
Spherocvtosis
Thalassemia

DECREASED ESR

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Anemia
Macrocytosis

INCREASED ESR

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Refrigerated sample not
returned to room temperature

INCREASED ESR

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Clotted blood sample
Delay in testing

DECREASED ESR

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Bubbles in ESR column
Low room temperature
Narrow ESR column diameter

DECREASED ESR

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High room temperature
Tilted ESR tube
Vibration

INCREASED ESR

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LEUKOCYTOSIS

DECREASED ESR

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LEUKEMIA

INCREASED ESR

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Hyperalbuminemia
Hyperglycemia
Hypofibrinogenemia
Hypogammaglobulinemia

DECREASED ESR

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Overanticogulation with EDTA causes a falsely _____ ESR

LOW

If ESR stands for more than 60 minutes, the results will be _______

FALSELY ELEVATED

If the test is timed for less than 60
minutes, invalidly ______ results are obtained

LOW

EOSINOPHILS
Highest ______
Lowest ______

Eosinophilia:
Eosinopenia:

Highest during the NIGHT
Lowest in the LATE MORNING

Eosinophilia: Allergic reactions, parasitic infestations, brucellosis, certain leukemias

Eosinopenia: hyperadrenalism, (Cushing’s disease), shock, ACTH administration

DIRECT METHOD- same as RBC and WBC count

______ present in the diluting fluid stains the eosinophils red

______ help to lyse the WBCs (except eosinophils)

______ lyses RBCs

______ present helps prevent clumping of WBCs

Phyloxine

Carbonate and water

Propelene glycol

Heparin

INDIRECT METHOD

A WBC count is performed on a specimen of blood
Make two blood smears and stain
Perform a 200-cell differential count on the blood smear

FORMULA: Eosinophils/ L= (%) Eosinophil in differential × WBC/L

SICKLE CELL ANEMIA

0% HbA
Glutamate ————> VALINE

SICKLE CELL TRAIT

60% HbA

Most common cause of death in sickle cells?

INFECTIOUS CRISES

deoxygenates hemoglobin
Hemoglobin S present in the red cell causes the formation of sickle shaped red cells
Cannot differentiate between sickle cell anemia and trait

SODIUM METABISULFITE METHOD

Interpretation: sickle cells or holly-leaf forms must come to a point/s to be considered positive

Red blood cells immediately lyse in the presence of saponin

HbS and other sickling hemoglobins, in the reduced state, form liquid crystals and yield a turbid appearance

SODIUM DITHIONITE TEST/ SOLUBILITY TEST

Hemolyzes the RBCs using a sample conditioner reagent
The Hb A and Hb S reagents contain monoclonal antibodies (IgG), which specifically binds to amino acids at or near the sixth position of the globin chain of hemoglobin A and hemoglobin S

Hemocard Hb A and S Procedure

HEMOCARD

RED or PINK: ______
GREEN, WHITE, GRAY:____

RED or PINK: POSITIVE
GREEN, WHITE, GRAY: NEGATIVE

Hb A (+)
Hb S (+)

SICKLE CELL TRAIT

Hb A (-)
Hb S (+)

SICKLE CELL ANEMIA

CELLULOSE ACETATE

A FAT SANTA CLAUSE

A>F>S>C

CITRATE AGAR

C>S>A>F

A screening test for Paroxysmal Nocturnal Hemoglobinuria (PNH)

Specimen:
Positive result:

SUGAR WATER SCREENING TEST

Specimen: CITRATED BLOOD
Positive result: HEMOLYSIS

A confirmatory test for PNH when the sugar water test is positive

Specimen:
Positive result:

SUCROSE HEMOLYSIS TEST

Specimen: CITRATED BLOOD
Positive result: HEMOLYSIS

ACID SERUM TEST

Ham’s method

Interpretation:
Normal- no lysis in any tube
PNH- hemolysis

NOTE: When the patient has received blood transfusion, less lysis occurs because of the presence of normal transfused red blood cells

A test to measure the ability of the red cell to take up fluid without lysing.

The primary factor affecting the red the osmotic fragility test is the shape of the red cell. which, in turn, depends on the volume, surface area, and functional state of the RBC membrane

OSMOTIC FRAGILITY TEST

SPHEROCYTES
HEMOLYTIC ANEMIA

Increased OFT (less resistant) marupok

SLS ITT
1. Splenectomy
2. Liver disease
3. Sickle cell anemia
4. IDA
5. Thalassemia
6. Target cells

Decreased OF (more resistant)

Reticulocytes have a ______ OFT

Decreased

SANFORD METHOD

________ is used as a diluent
12 tubes labeled 14-25
Number corresponds to the drops of 0.5% saline
Equate number of drops to 25 in all tubes
Factor used is ______

0.5% Saline

0.02

NV: initial hemolysis at tube 22 and complete hemolysis at tube 17

Example:

I.H= 20
C.H= 15 ———-> ⬇️ OFT (more resistant)

I. H= 24
C. H= 20——-> ⬆️ OFT (less resistant)

Tests for capillary abnormality that may be due to a defect in the capillary walls or to some type of thrombocytopenia
• Occasionally abnormal in hemophilia and Vit. K disorders

CAPILLARY FRAGILITY TEST

Blood pressure cuff is applied to the upper arm
Pressure applied should be between systole and diastole (100 mmHg for males; 80 mmHg for females)
After 5 minutes examine for petechia formation
First examination site should not be repeated within 7-14 days

Positive pressure test/ Rumple-Leede/ Tourniquet test

A 2 cm suction cup is used
Mid portion of the upper arm is used
The suction cup is applied for 1 minute
Pressure applied is 200-250 torr
Count petechiae 5 minutes after release

Negative pressure test/ Hess test/ suction test

A test the measures the Time it takes for a standard wound to stop bleeding.

Tests for:
Abnormalities of platelet function and number
VWF deficiency
Abnormality of vessel wall structure

BLEEDING TIME

15-20 mm above rounded fatty portion of earlobe
Uses a no. 11 sterile Bard-Parker surgical blade

N.V.:

MODIFIED DUKE METHOD

N.V.: <8 minutes (2-4 minutes)

Volar surface of the forearm is used

Use a blood pressure cuff and apply 40mmHg of pressure

How many times to puncture ______

Collect blood using filter paper strips after 2 minutes and again after every 30 seconds

N.V.:

IVY METHOD

Normal values: 3-6 minutes

Incision: 9mm x 1mm deep
N.V: 6-10 minutes

Standardized Simplate Test

In the past, it was used as a screening test to measure all stages of intrinsic coagulation system and monitor heparin therapy

It is sensitive only to extreme factor deficiencies and is insensitive to high doses of heparin

Lee and White Coagulation Time

Lee and White PROCEDURE:

1ml is dispensed into three tubes (3,2,1)
Timer is started by the time blood is dispensed at tube 3
Incubate at _____ for ____ minutes using a water bath
Tube 1 is tilted every _____seconds at an angle of ____ to check for clotting
The time when tube has clotted is recorded, after 30 seconds tube 2 is checked for clotting until a clot has formed
Repeat the same procedure for tube 3
Clotting time of tube 3 is reported
N.V:

37°C; 5 minutes

every 30; 45°

5-15 minutes

Stains siderotic granules, Pappenheimer bodies and hemosiderin

Prussian Blue reaction

Stains ALP present in the neutrophil but not in monocytes

Helpful in differentiating CML from leukemoid reaction or polycythemia vera

Leukocyte Alkaline Phosphatase

Polycythemia vera
last trimester of pregnancy
infections with neutrophilia

INCREASED LAP

CML, PNH, sickle cell anemia, IM, PA

Decreased LAP

Normal values in LAP

30-185

Mature neutrophils and bands are the only graded cells.

Score ranges from 0-4+

Specimen for LAP

Fresh capillary blood is recommended

Used to differentiate acute myelogenous leukemia(AML) and monocytic leukemia from acute lymphocytic leukemia (ALL)

Granulocytes (+)
Monocytes (-)

MYELOPEROXIDASE STAIN

Used to differentiate acute myelogenous leukemia and myelomonocytic leukemias from acute lymphocytic leukemia

Granulocytes (+)
Monocytes (-)

SBB

Stains mucoproteins, glycoproteins, and high molecular weight carbohydrates

Does not stain carbohydrates on pronormoblasts
Used to help in the diagnosis of DiGuglielmo’s syndrome (FAB M6)

PERIODIC ACID SCHIFF STAIN

Stains esterases in granulocytes
Used to differentiate granulocytic cells (+) from monocytic cells (-)

CHLOROACETATE ESTERASE/ SPECIFIC ESTERASE

Stains esterases present in the monocytic cells, macrophages, megakaryocytes, and platelets

Used to differentiate monocytic (+) leukemias from granulocytic (-) leukemias

NONSPECIFIC ESTERASE

stain is helpful in diagnosing hairy-cell leukemia (B-cell malignancy)

Tartrate-resistant ACP

Useful for the recognition of mast cells and tissue basophils

Toluidine blue

Screening procedure for the detection of chronic granulomatous disease (CGD), a phagocytic defect and NADPH OXIDASE deficiency

Nitroblue Tetrazolium Neutrophil Reduction Test
- reverse result

(-): with CGD
(+): no CGD

Stains DNA polymerase
Present in 90% cases of ALL
Used to differentiate AML from ALL
Stains lymphoblast

Terminal Deoxyribonucleotidyl Transferase

Requires fresh blood specimen

LAP
MPO

Begins during the embryonic development in blood islands of the yolk sac at around _______ of gestation

Mesoblastic Phase (Yolk Sac Phase)

19 days

Primitive erythroblasts arise from mesodermal cells

produce hemoglobin (Portland, Gower-1, Gower-2)

This phase of hematopoiesis occurs intravascularly

Begins at 4th to 5th gestational weeks

Characterized by clusters of developing erythroblasts, granulocytes, and monocytes

Start of definitive hematopoiesis

Lymphoid cells begin to appear

Liver is the major site of hematopoiesis and retaining activity until 1-2 weeks after birth

The spleen, kidney, thymus, and lymph nodes contribute to the hematopoietic process

DETECTABLE levels of hemoglobin (Hb) F, Hb A, and Hb A2 may be present

HEPATIC PHASE
- start of hematopoiesis

At 5th month development, hematopoiesis begins in the BONE MARROW

M:E ratio reaches adult levels of 3:1 at 21 weeks of gestation

MEASURABLE levels of Hb A1, fetal hemoglobin, and Hb A2

After the first 3 weeks postpartum, the bone marrow becomes the only normal site of blood cell production and remains so throughout life

MEDULLARY PHASE

TWO TYPES OF BONE MARROW

Red Marrow
Yellow marrow

Active bone marrow

At 18 years of age, the only active hematopoietic sites are:

Red marrow

Sternum
skull
vertebrae
ribs
pelvis
proximal extremities of long bones

hematopoietically inactive, comprised of adipocytes

YELLOW MARROW

The process of replacing the red marrow. by the yellow marrow is called ______.

RETROGRESSION

It is capable or reverting back to active marrow in cases of increased demand.

Approximately, there is equal amount of red and yellow marrow in adults

Significant role in hematopoiesis in the 2nd trimester and the major site during hepatic stage

Capable of extramedullary hematopoiesis (counter part of hepatic phase in adults) in case of bone marrow shut down

LIVER

Removes senescent RBCs
Sequesters approximately 30% of platelets

SPLEEN

3 types of tissues:

a. White pulp- consists of scattered follicles with germinal centers containing lymphocytes, macrophages, and dendritic cells
b.
Marginal zone- forms a reticular meshwork containing blood vessels, macrophages, and specialized B-cells
C.
Red pulp- comprised on dendritic processes that create a filter, the cords of Billroth, stagnates and depletes the glucose supply of RBCs that lead to their removal

cells are phagocytosed with subsequent degradation of cells and organelles

Culling

splenic macrophages remove inclusions or damaged surface membrane form RBCS

Pitting

All blood cells are derived from a single pluripotential stem cell.

More accepted theory

MONOPHYLETIC THEORY

Suggests that each of the blood cell lineages is derived from its own unique
stem cell

POLYPHYLETIC THEORY

Common lymphoid Progenitor

T cell
B cell
NK cell

Common Myeloid progenitor

Granulocytic
Erythrocytic
Monocytic
Megakaryocytic lineage

A calculated value used to establish the number of cells undergoing mitosis, normally it is 1-2%

MITOTIC INDEX

⬆️ MI= increased proliferation except in megaloblastic anemia wherein mitosis is prolonged

RNA and PROTEIN synthesis
10 hrs

Synthesis (S) phase

DNA synthesis
9 hours

Premitotic stage
4 hrs

Mitosis

Cell division
IPPAT

1 hour

Rest stage
Quiescence
Limbo
Repair

glycoproteins that REGULATE the proliferation, differentiation, and maturation of hematopoietic precursor cells

CYTOKINES

IL 1,3,6,9,11, GM-CSF, and Kit-ligand
Transforming growth factor B, TNF-a, Interferons

Positive influence- would like cells to proliferate
Negative influence- inhibition

Normoblastic: PRONORMOBLAST
Rubriblastic:
Erythroblastic:

Normoblastic: PRONORMOBLAST
Rubriblastic: RUBRIBLAST
Erythroblastic: PROERYTHROBLAST

Normoblastic:
Rubriblastic: PRORUBRICYTE
Erythroblastic:

Normoblastic: BASOPHILIC NORMOBLAST
Rubriblastic:
Erythroblastic: BASOPHILIC ERYTHROBLAST

Normoblastic:
Rubriblastic:
Erythroblastic: POLYCHROMATIC ERYTHROBLAST

Normoblastic: POLYCHROMATIC NORMOBLAST
Rubriblastic: RUBRICYTE
Erythroblastic:

Normoblastic:
Rubriblastic: METARUBRICYTE
Erythroblastic:

Normoblastic: ORTHOCHROMIC NORMOBLAST
Rubriblastic:
Erythroblastic: ORTHOCHROMIC ERYTHROBLAST

Normoblastic:
Rubriblastic: RETICULOCYTE
Erythroblastic:

Normoblastic: POLYCHROMATOPHILIC EEYTHROCYTE
Rubriblastic:
Erythroblastic: POLYCHROMATOPHILIC ERYTHROCYTE

Last RBC development stage

ERYTHROCYTE

Nucleus takes up much of the cell (high N:C ratio)
Measures 14-20 um and cytoplasm is quite blue
GLOBIN production begins

PRONORMOBLAST (Rubriblast)

n:c ratio decreases to 6:1
nucleoli usually not visible
measures 12-17 um and the cytoplasm stains deep blue
DETECTABLE level of HEMOGLOBIN synthesis (minute amount)

BASOPHILIC NORMOBLAST (Prorubicyte)

N:C ratio is 4:1

Measures 10-15 um and the cytoplasm is pink blue (murky-gray blue)

This is the last stage capable of
MITOSIS

1st stage where Hb synthesis is
VISIBLE

POLYCHROMATIC NORMOBLAST (rubricyte)

The nucleus is pyknotic

Pink-orange color of the cytoplasm reflects nearly complete production of hemoglobin

Later in this stage the NUCLEUS is ejected

Orthochromic normoblast (metarubricyte)

No nucleus

Cytoplasm is the predominant color of hemoglobin (pink)

Called a reticulocyte when the remnants of the ribosomal

RNA (reticulum) are stained with supravital stain (E. G. Nmb)

Polychromatophilic erythrocyte (Reticulocyte)

No nucleus

ave 7.5 um

Biconcave disc measuring 7-8 um in diameter

Appears salmon pink or red with a pale central palor

Has a lifespan of 120 days

ERYTHROCYTE

ERYTHROKINETICS

Erythron- collection of all stages of erythrocyte throughout the body

RBC mass- cells in the circulation

Hypoxia is detected by peritubular interstitial cells, which produces EPO

EPO- a glycoprotein hormone which is the major stimulatory cytokine for RBC
——-Early release of reticulocytes
——-Inhibition of apoptosis
——-Reduced marrow transit time

Anaerobic glycolytic pathway

Results in a net gain of 2 ATP molecules per 1 glucose molecule

Generates 90% of RBC’s ATP

Common enzyme being deficient is
PYRUVATE KINASE.

Embden-Meyerhof pathway

Aerobic glycolysis

NADP+ is reduced to NADPH

NADPH reduces glutathione

Reduced glutathione reduces PEROXIDASE to water

Common enzyme that is deficient is G6PD.

Hexose monophosphate pathway

NADPH reduces the Ferric
to the Ferrous in the presence of methemoglobin reductase

Methemoglobinreductase pathway

Generates 2,3-DPG regulates oxygen delivery to tissues by competing with oxygen for hemoglobin

When 2-3 DPG binds hemoglobin, oxygen is released which enhances delivery of oxygen to tissues

Luebering-Rapaport Shunt

Mechanical hemolysis/ Intravascular hemolysis.

____ and ____ salvage release hemoglobin so that iron is not loss in the urine

HAPTOGLOBIN and HEMOPEXIS

Pathologic processes that produce changes in the exterior membrane of the RBCs causes premature removal by the macrophages

Excessive extravascular hemolysis

E.g. Heinz bodies, intracellular parasites, immunoglobulins or complement on BC membrane
Increased in plasma unconjugated bilirubin with subsequent increase of urobilinogen that is excreted by the kidneys due to increased presentation of unconjugated bilirubin to the liver

Traumatic physical lysis of RBCs caused by prosthetic heart valves, malarial parasites.

Hemoglobinemia, hemoglobinuria, hemosiduria and eventual increase in urinary urobilinogen

Decreased haptoglobin and hemopexin

Excessive Intravascular Hemolysis

Serum

⬇️ haptoglobin
⬇️ hemopexin

INTRAVASCULAR

URINE

⬆️ UROBILINOGEN
HEMOGLOBINURIA

⬆️ UROBILINOGEN
——- intravascular and extravascular
HEMOGLOBINURIA
——-intravascular

WHOLE BLOOD

⬇️hb, hct, RBC
⬇️ glycated hemoglobin

INTRAVASCULAR
EXTRAVASCULAR

Synthesis Occurs in the mitochondria and cytoplasm of bone marrow RBC precursors.

Begins with condensation of ______ and _________ catalyzed by _________ form aminolevulinic acid.

HEME

glycine and succinyl coenzyme A; aminolevulinic acid synthase

In the presence of _________ catalyzes the formation of porphobilinogen

ALA dehydratase (porphobinogen synthase)

In its presence, it catalyzes formation of hydroxymethylbilane

POPHOBILINOGEN DEAMINASE (hydroxy-methylbilane synthase)

Fe2 combines with protoporphyrin IX in the presence of ________ to make heme.

Ferrochelatase/ heme synthase

Rate limiting step

Glycine+Succinyl CoA

Causes hereditary sideroblastic anemia

ALA synthase

Contributes in LEAD poisoning

ALA dehydratase
Ferrochelatase

A.k.a HEME SYNTHASE

Ferrochelatase

Produced on specific ribosomes in the cytoplasm of RBCs

GLOBIN

Globin codes in Chromosome 11

Gamma
Beta
Delta
Epsilon

Globin chain that codes in Chromosome 16

Alpha
Zeta

Globin that has 2 genes

Beta
Delta
Zeta
Epsilon

4 genes

Gamma
Alpha

Hemoglobin: Gower I
Molecular Structure:
Stage of life:

Hemoglobin: Gower I
Molecular Structure: 2 zeta, 2epsilon
Stage of life: EMBRYONIC

Hemoglobin:
Molecular Structure: 2 alpha, 2 delta
Stage of life:

Hemoglobin: A2
Molecular Structure: 2 alpha, 2 delta
Stage of life: NEWBORN & ADULT

Hemoglobin: GOWER II
Molecular Structure:
Stage of life:

Hemoglobin: GOWER II
Molecular Structure: 2 alpha, 2 epsilon
Stage of life: EMBRYONIC

Hemoglobin:
Molecular Structure: 2 alpha, 2 gamma
Stage of life:

Hemoglobin: Fetal (HbF)
Molecular Structure: 2 alpha, 2 gamma
Stage of life: NEWBORN & ADULT

Hemoglobin:
Molecular Structure: 2 alpha, 2 beta
Stage of life:

Hemoglobin: A1
Molecular Structure: 2 alpha, 2 beta
Stage of life: NEWBORN & ADULT

Hemoglobin: PORTLAND
Molecular Structure:
Stage of life:

Hemoglobin: PORTLAND
Molecular Structure: 2 zeta, 2 gamma
Stage of life: EMBRYONIC

NORMAL HEMOGLOBIN CONCENTRATION IN ADULTS

92-95% HbA
2-3% HbA2
1-2% HbF

Describes the relationship between pO2 and the oxygen content of hemoglobin

Hemoglobin has a low affinity for oxygen at low oxygen tension and a high affinity for oxygen at high oxygen tension

OXYGEN DISSOCIATION CURVE

Shifts of curve to the left or right occur if there are changes in the pH of blood

BOHR EFFECT

Increased affinity for oxygen leading to a decreased oxygen delivery

SHIFT TO THE LEFT

Caused by an increase in:

CO2
Acidity (an increase in H*)
2-3 DPG
Etcetera (Hb variants with decreased affinity for 02)
Increased temperature

Shift to the RIGHT

Lowered body temperature
Blood transfusion with depleted 2-3 DG
Alkalosis
Methemoglobinemia
Increased carboxyhemoglobin
Some Hb variants

Shift to the LEFT

Alkalinity (a decrease in H*)
Decrease 2-3 DPG
Decreased PCO2
Decreased temperature
b variants with increased affinity for 02

Shift to the LEFT

High fever
Acidosis
Conditions that produce hypoxia

Shift to the RIGHT

15-20um
Deeply basophilic cytoplasm, round or oval nucleus
N:C ratio of 4: 1

MYELOBLAST

9-15um

Pink to rosy violet granules

Neutrophilic granules contain ACP, acid hydrolase, muramidase, and lactoferrin (binds iron to inhibit bacterial growth) which are essential for phagocytosis.

Nucleus has 2-5 lobes (>6 lobes indicates shift to the right)

Hypersegmentation is seen in vit B12 deficiency

POLYMORPHONUCLEAR NEUTROPHIL

15-21um
Deeply basophilic cytoplasm
Appearance of PRIMARY GRANULES
N:C ratio of 3:1 to 2:1

PROMYELOCYTE

9-15 um
Curved nucleus/ sausage shaped

NEUTROPHILIC BAND

Appearance of SECONDARY GRANULES
Nucleus is SLIGHTLY INDENTED (D- shaped)
N:C ratio= 1:1
Last stage capable of MITOSIS

1st stage that allow granulocyte differentiation

MYELOCYTE

10-15 um
Kidney shaped nucleus
aka JUVENILE CELLS

METAMYELOCYTE

MITOTIC/ PROLIFERATING POOL (Circulating pool)

HSC, CMP, CFU-GEMM, GMP, MYELOBLAST, PROMYELOCYTE, MYELOCYTE

STORAGE POOL (Marginating Pool)

Metamyelocyte, Band, Segmented Neutrophil

MPO, cathepsins, defensins, elastase, proteinase 3, acid-b-glycerophosphate

PRIMARY (AZUROPHILIC)
PROMYELOCYTE STAGE

b2-Microglobulin, Collagenase, Gelatinase, Lactoferrin, lipocalin,
Transcobalamin

SECONDARY (SPECIFIC)
MYELOCYTE AND METAMYELOCYTE

Gelatinase, Collagenase, Lysozyme, Acetyltransferase, B2 microglobulin

TERTIARY
METAMYELOCYTE AND BAND

Eosinophilic granules contains:

contain peroxidase, ACP, and other proteolytic enzymes, but do not contain
ALP

Remnants of Eosinophil

CHARCOT LEYDEN CRYSTALS

Basophilic granules contains:

histamine, heparin, and chondroitin sulfate

Mast cells originates from ______
Basophils originates from _____

C.T mesenchyme
HSC

14-20 um
Blue gray cytoplasm
Many fine, azurophilic granules giving the cell a characteristic “ground glass appearance”
Nucleus is horse-shoe shaped/ kidney shaped

MONOCYTE

MITOTIC POOL 2-3 davs
STORAGE POOL 5-7 days
LIFESPAN: 9-10 days from myeloblast to death

NEUTROPHIL

MATURATION: B-cell: 30-36 hours
LIFESPAN: some live for 3-4 days, majority live for months to years.

LYMPHOCYTE

Maturation: 3.5 days
Half life: 18 hours

EOSINOPHIL

Maturation and storage: 4.3 days or 12 hours
Transit time in the peripheral blood: 3.7 days

BASOPHIL

Maturation: 30-48 hours (60hrs) nodales
TISSUE PHASE: macrophage: months, possibly longer.
Few hours (inflammatory macrophage)

MONOCYTE

PLATELETS

Anucleate blood cells
NV: 150-450 x 10^9/L
Approximately _____ per OIF
Life span: _____ days
Average diameter: _____ um
Mean platelet volume of _____fl
_____ is sequestered in the spleen
On a Wright-stained wedge-preparation blood film, platelets appear (_________ to irregular, lavender, and granular

Anucleate blood cells
NV: 150-450 x 10°/L
Approximately 7-21 per OIF
Life span: 8-11 days
Average diameter: 2.5 um
Mean platelet volume of 8-10 fl
20-30% is sequestered in the spleen
On a Wright-stained wedge-preparation blood film, platelets appear (CIRCULAR to irregular, lavender, and granular

Endomitosis: No cytokinesis and telophase (no daughter cells)

MEGAKARYOCYTOPOIESIS

RETICULATED PLATELETS sometime known as:

Stress
Thrombocytopenia
Immature

20-60 um
Less basophilic cytoplasm
Nucleus is IRREGULARLY shaped

First stage where DEMARCATING SYSTEM appear

PROMEGAKARYOCYTE

MEGAKARYOCTE LINEAGE COMMITED
PROGENITORS

BFU-MEG- least mature, capable of mitosis
CFU-MEG- intermediate, capable of mitosis
LD-CFU-MEG- most mature, ENDOMITOSIS

Metamegakaryocyte

Cytoplasmic granules:
Cytoplasmic tags:
Nuclear features:
Thrombocyte visible:

Cytoplasmic granules: Aggregated
Cytoplasmic tags: Absent
Nuclear features: Four or more nuclei
Thrombocyte visible: YES

Cytoplasmic granules: ABSENT
Cytoplasmic tags: PRESENT
Nuclear features: SINGLE NUCLEUS, fine chromatin, nucleoli
Thrombocyte visible: NO

MEGAKARYOBLAST

Nucleus: MULTILOBED
Nucleoli: NOT VISIBLE
Chromatin: deeply but variably condensed
Demarcation system: PRESENT

MK-III (MEGAKARYOCYTE)

Nucleus: ROUND
Nucleoli: 2-6
Chromatin: HOMOGENOUS
Demarcation system: PRESENT

MK-I (MEGAKARYOBLAST)

Nucleus: INDENTED
Nucleoli: VARIABLE
Chromatin: condensed
Demarcation system: PRESENT

MK-II (PROMEGAKARYOCYTE)

What stage does proplatelet process appear?

MEGAKARYOCYTE

If the megakaryocyte has more nuclear lobes, platelet production will be _______

INCREASED

MPV is ______ related to platelet count

INVERSELY

Which of the following has demarcating system?

I. MK-I
II. MK-II
III. MK-III

All of the following

Where glycoproteins gp1b and gp2b3a are found
Plasma membrane
Submembranous area

PERIPHERAL zone

Glycocalyx

Microfilaments (actin+myosin)
_______- contractile elements
_______-retains platelet shape

SOL GEL ZONE

Thrombosthenin/actomyosin

Microtubules (tubulin)

Where alpha granules and dense granules located

Organelle zone

________- arachidonic acid metabolism, activation system
Surface Connecting System- granules release

MEMBRANOUS SYSTEM

Dense Tubular System

Dense granules

ADP- AGGREGATION
ATP
SEROTONIN- FOR VASOCONSTRICTION
Ca, Mg- support platelet activation and coagulation
Pyrophosphate

The dense granule contents are vasoconstrictors and platelet agonists that amplify primary hemostasis;

B-thromboglobulin
Factor V
Factor IX
Protein S
Fibrinogen
VWE
Platelet factor 4
Platelet derived growth factor

Alpha granules

most of the a-granule contents are coagulation proteins that participate in secondary hemostasis

Platelet roll and cling to nonplatelet surfaces
Reversible
Seals endothelial gaps
Some secretion of growth factors

Plt. ADHESION

platelets adhere to each other
Irreversible
Platelet plugs form
Secretion of all platelet contents
Requires fibrinogen

Platelet AGGREGATION

Irreversible
Occurs during aggregation
Essential to coagulation

Plt. SECRETION

A spectrophotometric approach to hemoglobin measurement.
A. Blood oxygen capacity
C. Cyanmethemoglobin
B. Blood Iron content
D. Copper sulfate method

Which of the following components of the modified Drabkin’s reagent removes the interference from abnormal proteins?
A. Potassium cyanide
B. Potassium ferricyanide
C. KH2PO4
D. Nonionic detergent

Which of the following is error is correctly matched with its remedy?
A. Elevated WBC: Dilute mixture 1:1 with water then multiply results by 2
B. Hemoglobin S and C: centrifuge the mixture then use the supernatant
C. Lipemic blood: Add 01 mL patient’s plasma to 5.0 mL of HiCN and use as blank
D. Abnormal globulins: Add potassium oxalate to Drabkin’s reagent

Which of the following statements is CORRECT?
A. Potassium ferricyanide donates the cyanide group to hemoglobin
B. The color intensity is inversely proportional to hemoglobin concentration in cyanmethemoglobin method
C. Optical density is measured at 540 nm
D. Potassium cyanide combines with hemoglobin to form methemogloblin

The blood oxygen capacity is based on the fact that 1g of Hb carries
A. 0.347 C. 1.053
B. 1.34 D. 0. 134

A patient suffering from carbon monoxide poisoning will be characterized by?
A. A cherry red discoloration of the face
B. A mauve lavender color of the blood
C. A chocolate brown discoloration of the blood
D. Highest absorbance observed at 618 pm

All of the following are true regarding sulfhemoglobin EXCEPT?
A. Formed by the action of certain drugs such as aromatic amines
B. Absorbance at 540 m
C. Not measured by cyanmethemoglobin method
D. Imparts a mauve lavender color to the blood

Which of the following is correctly matched?
A. Red band- contains heparin: anticoagulated sample
B. Blue band- heparin: non-anticoagulated sample
C. Red band- plain tube: anticoagulated sample
D. Blue band: plain tube: anticoagulated sample

Which of the following is/are true regarding microhematocrit method?
I. 75 mm long with 1.2 mm internal bore
II. Centrifugation done at 5000-10000 RCF
III. Centrifugation time is 10 minutes
IV. Clay plug is at least 4-6 mm long

A. 1. 11. I/
B. 1& IlI
C. I & IV
D. I. IL. IIL IV

Which of the following cause/s a falsely decreased microhematocrit resuts?
I. Incomplete sealing
Il. Incomplete centrifugation
Ill. Over anticoagulation
IV. Trapped plasma

A. I,II,III
B. I& III
C. II & IV
D. I, II, III, IV

A medical technologist left a microhematocrit tube to stand for a long period of time before reading, what is expected to happen with the results?
A. Falsely decreased hematocrit
B. Falsely increased hematocrit
C. No effect
D. Either A or B

Which of the following is correct for the Wintrobe tube?
A. 300 mm long
B. Left side is used for macrohematocrit
C. Right side is calibrated from 100-0
D. Right side is used for microhematocrit

Diluting fluids used in RBC count EXCEPT:
A. Dacie
B. Eagle
C. Bethel
D. Turk’s

A neutrophil that has a curved or sausage-shaped nucleus
A. Segmenter
B. Band
C. Juvenile
D. PMN

The youngest WBC that is normally present in the differential count.
A. Segmenter
B. Juvenile
C. Neutrophil metamyelocyte
D. Band

The first stage that allows granulocte differentiation.
A. Segmenter
B. Promyelocyte
C. Myelocte
D. Metamyelocyte

Which of the following do/does not produce monocytosis?
I. Tuberculosis
II. Brucellosis
III. EBV
IV. Whooping cough
A. 1 & 11
B. Ill & IV
C. I & III
D. II & III

The optimal EDTA concentration for anticoagulation is
A. 2.0 mg/mL
B. 1.0 mg/mL
C. 1.5 mg/mL
D. 2.5 mg/MI

The anticoagulant of choice for OFT.
A. Heparin
B. Citrate
C. EDTA
D. Oxalate

The exact measurements of the needle that is most commonly used for routine venipuncture.
A. 21 gauge, 1.5 inches
B. 20 gauge, 1.5 inches
C. 21gauge, 1.0 inches
D. 20 gauge, 20 inches

Due to a difficulty encountered in the venipuncture procedure, only 0.5 mL out of 2 mL was collected in an EDTA tube, this will lead to?
I. Decreased ESR
II. Decreased hct
III. Decreased RBC
IV. Decreased Plt and WBC

A. I, II, III, IV
B. I, II, II
C. I & II
D. 1 & III

A medical technology intern expels blood through the needle from a syringe into an EDTA tube, this would lead to:
I. Decreased RBC
II. Increased Hct
Ill. Decreased MCV
IV. Increased MCHC

A. I, II, III, IV
B. II & I
C. I & IV
D. I, III & IV

The manual reticulocyte count values should agree within?
A. 5%
B. 10%
C. 15%
D. 20%

The reticulocyte count is performed under the OlF. The miture should be allowed to stand for 10 minutes before counting is performed.
A. First statement is correct
B. Second statement is correct
C. Both are correct
D. Both are incorrect

For how long should RBCs be lysed by the lytic agent before WBC count is performed?

A. 5 minutes
B. 10 minutes
C. 15 minutes
D. 20 minutes

Platelets are tiny refractile elements that arise from the cytoplasm of megakaryocytes. The shape of these cells in a Wright-stained smear is round.
A. First statement is correct
B. Second statement is correct
C. Both are correct
D. Both are incorrect

Shift to the right of WBCs is seen in conditions wherein DNA synthesis is impaired. A shift to the right of RBCs is seen in conditions wherein DNA synthesis is impaired.
A. First statement is correct
C. Both are correct
B. Second statement is correct
D. Both are incorrect

Platelet progenitor cells undergo chromosomal replication without cell division. The megakaryocytic system becomes smaller as the cell matures.
A. First statement is correct
C. Both are correct
B. Second statement is correct
D. Both are incorrect

This WBC is difficult to see in the manual WBC differential because of the water solubilitv of its
granules.
A. Neutrophils
B. Monocytes
C. Basophils
D. Eosinophils

Mitotic pool in the bone marrow is comprised of?
A. HSC to neutrophil segmenter
B. Myeloblast to myelocyte
C. HSC to myelocte
D. CMP to metamyelocyte

Which cytokine negatively affects cell proliferation?
A. IL- 1
B. TGF-B
C. IL-9
D. IL- 11

Reticulocytes that stay longer than 1 day in the circulation will contribute more to the reticulocyte count. Reticulocytes have blue granulofilamentous materials that are demonstrated using Wright’s stain
A. First statement is correct
B. Second statement is correct
C. Both are correct
D. Both are incorrect

Dendritic cells are believed to arise from?
A. Common lymphoid progenitor
B. Monocytes
C. Lymphocytes
D. T-cells

A stain that is used to differentiate FAB M6 (acute erythroleukemia) from ALLs.
A. Toluidine blue
B. PAS
C. TDT
D. TRAP

Tiny B lymphocytes that have fine cytoplasmic extensions possess.
A. Myeloperoxidase
B. Esterases
C. ACP 5
D. LAP

A WBC that has ALP.

A. Neutrophil
B. Monocyte
C. Lymphocyte
D. Eosinophil

Which of the following WBCs will be negative for esterase?
A. Neutrophil
B. Monocyte
C. Lymphocyte
D. Eosinophil

If the WBCs counted in 10 fields are, 16, 9, 4, 12, 7, 11, 5, 10, 9, 3. What would be the WBC estimate?
A. 18x109/L
B. 14.4×10°/L
C. 17.2×10°/L
D. 19X10 9/L

Platelet estimate is performed at______ and the average is multiplied by the factor ______
A. HPO, 10000
B. 0I0, 2000
C. HPO, 2000
D. 0I0, 20000

In the OFT procedure, NSS is used. The OFT method is called Sanford.
A. First statement is correct
B. Second statement is correct
C. Both are correct
D. Both are incorrect

Tests for PH EXCEPT:
A. Sugar water screening
B. Sucrose hemolysis
C. Sodium metabisulfite
D. Acid serum test

Megakaryocytic stage characterized by an irregularly shaped nucleus.
A. Megakaryoblast
B. Promegakaryocyte
C. Megakaryocytic
D. Metamegakaryocyte

Lifespan: 9-10 days from myeloblast to death.
A. Neutrophil
B. Monocyte
C. Lymphocyte
D. Eosinophil

Maturation 3.5 days, lifespan: 18 hours
A. Neutrophil
B. Monocyte
C. Lymphocyte
D. Eosinophil

Maturation: 30-48 hrs, lifespan: months
A. Neutrophil
B. Monocyte
C. Lymphocyte
D. Eosinophil

Maturation: 1-2 days, lifespan: months to years
A. Neutrophil
B. Monocyte
C. Lymphovte
D. Eosinophil

Shift to the left
I. Methemoglobinemia
II. Drowning
III. Carboxyhemoglobin
IV. Hypokalemia

A. I, II, III
B. I. II. IV
C. I, II, III, IV
D. I, III, IV

Which of the following will be increased in the morning?
I. Cortisol
II. ACTH
III. Fe
IV. Eosinophils

A. I, II, III
B. II, III
C. I, III
D. I, II

The rate limiting step in hemoglobin synthesis:
A. Condensation of glycine and succinyl CoA into ALA
B. ALA conversion to PBG by ALA dehydratase
C. PBG conversion to UPG II!
D. Coupling of iron to PIX

The precipitation of Heinz bodies is indicative of?
A. A deficiency in EMP
B. A deficiency in HMP
C. A defect in MetHb reductase pathway
D. A defect in LRS

All of the following are true regarding microhematocrit determination EXCEPT:
A. Tube is 75mm long by with 1.2 mm bore.
B. Can hold 0.5 ml of blood
C. EDTA can decrease results
D. Trapped plasma makes results higher

What dilution should be used if the WBC count is 40×109/L?
A. 1:10
B. 1:11
C. 1:100
D. 1:200

The smear that is created by a medical technology intern is excessively blue both macroscopically and microscopically, what can be done to correct this?
A. Increase the angle of spreader
B. Decrease the pressure
C. Shorten washing time
D. Decrease size of blood drop

The color of a properly made smear when put directly under light should give a ________.
A. Rainbow shine
B. Blue color
C. Red color
D Red and blue color

A stain that differentiates acute myelogenous leukemia and acute monocytic leukemia from ALL
A. MPO
B. SBB
C. PAS
D. Specific esterase

Which of the following statement/s is/are true
Trapped plasma can cause an elevated hematocrit reading using hematology analyzers
II. Trapped plasma is found in the fourth layer of a spun blood.
III. Sequestrene can cause shrinkage of RBCs leading to a falsely decreased hematocrit reading
IV. Plain microhematocrit tubes should be used to contain non-anticoagulated samples

A. 1& I
B. 1& IV
C. Il & IlI
D. II & IV

The most commonly used system for venipuncture.
A. Syringe
B. Evacuated tube
C. Butterfly needle
D. Lancet

Which of the following statements is/are INCORRECT.
I. The angle of needle insertion for routine venipuncture is 15°-30°
II. The angle of needle insertion for venipuncture is 25°-40°
III. The most commonly used needle is the 21 gauge needle
IV. The higher the needle gauge, the larger the needle bore
A. I & II
B. II& III
C. I & IV
D II & IV

Which of the following is not a characteristic of the median cubital vein?
A. It is the largest vein, closest to the surface
B. It is the most stationary vein in the antecubital fossa
C. It is the easiest vein to palpate in obese patients
D. Most preferred vein

Which of the following veins are suitable for venipuncture?
I. Veins on the back of the hand
II. Veins on the back of the wrist
III. Median cubital vein
IV. Basilic vein
A. I, II, III
B. I, II, III, IV
C I, II, IV
D. I. IL. IV

What should a medical technologist do when a patient refuses to undergo venipuncture?
A. Inform the physician and let the patient sign a waiver stating that he/she refused to the procedure
B. Let the physician do the extraction
C. The medical technologist should gently persuade the patient
D. Let the attending nurse do the extraction

Which of the following shows the correct order of draw?
A. Blue-Yellow-Red-Green
B. Yellow-Black-Orange-Green
C. Yellow-Red-Blue-Green
D. Yellow-Lavender-Blue-Red

A stat blood chemistry was requested by a physician, what tube must be used by the medical technologist?
A. Red (glass)
B. Red (plastic)
C. Green
D. Orange

What is the primary reason why Na citrate is the preferred tube for coagulation tests?
A. It binds calcium
B. It preserves factor V and VIII
C. It prevents premature activation of clotting factors
D. It preserves factor VIl and XI

A gray top tube contains what anticoagulant?
A. Na fluoride
B. lodoacetate
C. Oxalate
D. Citrate

Which of the following is used to prevent the aggregation of platelets around neutrophils in a peripheral blood smear?
A. EDTA
B. Na citrate
C. Heparin
D. Oxalate

What results are to be expected when using a capillary blood?
I. Lower RBC
II. Lower Hct
III. Higher platelets
IV. Lower WBC count

A. 1 & II
B. I, II, III
C. I, II, III, IV
D. I & IV

Correct skin puncture procedure
A. Slide-Blood gas-EDTA-Gray
B. EDTA-Slide-Blood gas-Green
C. Blood gas-slide-EDTA-Green
D. Blood gas-slide-EDTA-Serum

What color stopper must be used for ESR?
A. Blue
B. Black
C. Lavender
D. Red

Which of the following statements is INCORRECT?
A. A patient who is standing up for a long time will have a relative increased in proteins and lipids
B. Cortisol and ACTH are high in the afternoon
C. Fe and eosinophils are high in the afternoon
D. Hb is increased in smokers

It is the layer in the spun blood where the trapped plasma is found.
A. 4th laver
B. 3rd layer
C. 2nd laver
D. 1st laver

CBC includes:
A. WBC, RBC, platelet count
B. Diff count, WBC, RBC, RBC indices
C. WBC, RBC, Hct, Hb, Diff count, RBC indices
D. WBC, RBC, Hct, Hb, Diff count, RBC indices, PIt.

Rule of three:
I. For QC and validity checks
II. RBC x 3= Hct +/- 3
III. For normocytic, normochromic BC
IV. 1 Hct= 107,000 RBCs/cumm

A. AOTA
B. Ill, IV
C. I. III. IV
D. I, II, III

Which of the following substances do not favour the adhesion of platelets onto the vascular endothelium?
A. 12 HETE
B. Gp1b
C. VWF
D. 13-HODE

A substance secreted by the endothelial cells that enhances the inhibitory effects of thrombin to factor V and VIll.
A. Thrombosthenin
C. Thrombomodulin
C. Thromboglobulin
D. Thrombospondin

It enhances AT-Ill activity in vivo.
A. Heparan sulfate
B. Adenosine
C. ATP
D. Serotonin

The enzyme that is inhibited by aspirin.
A. Lipoxygenase
B. Cyclooxygenase
C. Thromboxane synthetase
D. PGI2 synthetase

The protein which is responsible for maintaining the shape of the platelets.
A. Actin
B. Myosin
C. Actomyosin
D. Tubulin

Which of the following does not refer to HMWK?
A. Flaujeac factor
B. Williams Factor
C. Fletcher factor
D. Fitzgerald factor

Secreted by the dense granules.
A. Platelet cofactor 4
B. ATP
C. Thrombospondin
D. Thromboglobulin

Functions of Factor XIla. EXCEPT:
A. Initiation of fibrinolytic system
B. Conversion of prekallikrein to kallikrein
C. Production of kinins
D. Promotes thrombosis

Which of the following are found in serum?
A. Factor I
B .Factor lI
C. Factor V
D. VIII
E NOTA

Which of the following will lead to a falsely long clotting tests?
I.Moist needle
II. Overly filled Na citrate tube
IlI. Uncapped specimens
IV. Use of 3.2% Na citrate

A. I & II
B. III
C. I, II, III
D. II, IV

Which of the following will lead to a falsely shortened clotting tests?
I. Moist needle
II. Refrigeration
IlI. Hemolysis
IV. Use of 3.8% Na citrate

A. I & II
B. I, II, III
C. I, II, III, IV
D. I, II, IV

A test which uses thromboplastin as its reagent.
A. Prothrombin time
B. АРТТ
C. Plasma recalcification time
D. Lee White Clotting Time

Sensitive for factor VIII Deficiency
A. Prothrombin time
B. APTT
C. Plasma recalcification time
D. Lee White Clotting Time

An INR that is above 3.0 indicates:
A. Increased risk for haemorrhage
B. Increased risk for thrombosis
C. Normal
D. Increased fibrinolysis

An ISI that is close to 1 means that:
A. The sensitivity of the PT reagent is low
B. The sensitivity of the PT reagent is high
C. The PT reagent is specific
D. PT reagent is nonspecific

What condition could give the following lab results:
Thrombin time: Prolonged
Reptilase Time: Prolonged
A. Heparin therapy
B. Immunologic antithombins
C. Decreased fibrinogen
D. Abnormal globulins

It was used to help between factor X and VII deficiencies
A. Reptilase time
C. Duckhert’s test
B. Stypven time

A patient who has a history of bleeding has the following lab results:
Platelet count: 230x109/L
PT: 11 seconds (Control 12)
Bleeding time: 7 minutes
APTT: 50 seconds (control 35)
Thrombin time: Prolonged
Reptilase time: Normal

A. Liver disease
B. DIC
C. Heparin therapy
D. Coumarin therapy

Laboratory results of a patient are as follows:
Platelet count: 40 x10°/L
Bleeding time: 13 minutes
PT: 32 seconds (Control 12)
APTT: 60 seconds (control 35)
Thrombin time: Prolonged
A. The results will most likely correlate with a positive D-DIMER test
B. The results suggest that the patient has a deficiency in a factor that is involved in the common pathway
C. The results suggest that the patient has vWD
D. The results suggest pathologic fibrinolysis

What condition could produce the following results:
Platelet count: 160 x109/L
Bleeding time: 5 minutes
Thrombin time: Prolonged
PT: 20 seconds (Control 12)
APTT: 45 seconds (control 35)

A. DIC
B. Hemophila A
C. Liver disease
D. Christmas disease

What is the factor deficiency based on the data below.
PT: N
APTT: A
TT: N
NP: C
Adsorbed plasma: C
Aged serum: NC

A. Factor I deficiency
B. Christmas disease
C. Hemophilia A
D. Hemophilia C

Patient with a bleeding history gives the following results:
PT: N
APTT: A
TT: N
NP: C
Adsorbed plasma: C
Aged serum: C

A. Factor IX deficiency
B. Factor VIII deficiency
C. Factor XII deficiency
D. Factor XI deficiency

What condition could produce the following results:
Platelet count: 180 x10°/L
Bleeding time: 11 minutes
Thrombin time: Normal
PT: 12 seconds (Control 12)
APTT: 50 seconds (control 35)

A. DIC
B. Hemophila A
C. vWD
D. Christmas disease

Bleeding will not be prolonged in:
A. VWD
B. Bernard-Soulier syndrome
C. Glanzmann thrombasthenia
D. NOTA

A medical technologist observed platelets encircling neutrophils in peripheral blood smear, what corrective action should be done?
A. warm sample at 37C
B. Use Na citrate
C. Dilute sample
D. Continue to count the platelets

A platelet estimate of 50,000-99,000 should be reported as?
A. Markedly decreased
B. Moderately decreased
C. Slightly decreased
D. Low normal

Platelet count is <30,000/ uL.
A. Abnormal BT
B. Bleeding possible with trauma
C. Spontaneous bleeding possible
D. Severe spontaneous bleeding

Normal response to ristocetin, weak response to ADP, epinephrine and collagen.
A. Bernard-soulier
C. Hemophilia A
D. VWD
D. Glanzmann thrombasthenia

Vomiting of blood.
A. Hemoptysis
B. Hematoma
C. Hematemesis
D. Hematuria

Due to a peptidase enzyme deficiency.
A. Pseudoxanthoma elasticum
B. Ehlers-Danlos
C. Scurvy
D. Kassabach-Meritt Syndrome

Relationship of the number of blood units transfused to a patient’s degree of thrombocytopenia.
A. Indirect
B. Direct
C. Unrelated
D. Inverse

Platelet retention in paraproteinemias.
A. Increased
B. Decreased
C. Unaffected
D. Slightly increased

Which of the following conditions would lead to thrombocytopenia
I. Splenomegaly
II. Splenectomy
III. Leukoerythroblastic anemia
IV. CGL

A. I, II, III, IV
B. I, II, III
C. I & III
D. I & IV

A disease characterized by a triad of recurrent infections, thrombocytopenia and eczema.
A. WAS
C. Hermansky-Pudlak
B. Gray platelet syndrome
D. Chediak-Higashi

Thrombotic disorders (tendencies)
I. Plasminogen deficiency
II. Dysfibrinogenemia
III. Lupus anticoagulant
IV. Factor XII deficiency

A. I, II, III, IV
B. I, II, IlI
C. I & III
D. I& IV

The time it takes before D-dimer test gives a positive result.
A. after 2 hours
B. after 4 hours
C. After 8 hours
D. After 10 hours

Characteristic blood picture of acute blood loss anemia.
A. Microcytic, hypochromic
B. Normocytic. normochromic
C. Macrocytic, hypochromic
D. Macrocytic, normochromic

Which of the following does not produce a microcytic, hypochromic blood picture?
A. IDA
C. Thalassemia
B. ACD
D. Aplastic anemia

All of the following exhibit a normocytic, normochomic blood picture EXCEPT:
A. ACD
B. Lead poisoning
C. Hemolytic anemia
D. Thalassemia

Cells are more susceptible to lysis when exposed to a hypotonic solution.
A. Hereditary elliptocytosis
C. Hereditary spherocytosis
B. Hereditary stomatocytosis
D. Acanthocytosis

Associated with abetalipoproteinemia.
A. Hereditary elliptocytosis
B. Hereditary stomatoctosis
C. Hereditary spherocytosis
D. Acanthocytosis

Heinz bodies in RBCs indicate:
A. Membrane defects caused by polarization of cholesterol
B. Membrane defect due to an abnormal permeability to Na and K
C. Oxidation of ferrous iron to the ferric state
D. Decreased ATP production

Which of the following stains require a fresh sample?
I. LAP
IL. MPO
III. SBB
IV. PAS

A. I, II, Ill
B. I& III
C. I & II
D. II & III

Which of the following enzymes is only present in neutrophils?
A. Myelperoxidase
C. Chloroacetate esterase
B. ACP
D. LAP

After receiving blood transfusion, a patient with PH will exhibit:
A. Increased lysis for Ham’s method
C. Increased lysis for sucrose hemolysis test
B. Decreased lysis for Ham’s method
D. Increased lysis for sugar water test

Requires a fresh heparinized sample.
A. LAP
B. MPO
C. SBB
D. Specific esterase

The presence of a biphasic antibody causes hemolvsis in red blood cells
A. PNH
B. PCH
C. Spherocytosis
D. Acanthocvtosis

The poikilocyte that is has decreased osmotic fragility.
A. Spherocytes
B. Acanthocytes
C. Codocytes
D. Dacryocytes

Sodium dithionate test is a screening test for:
A. 6th amino acid substitution of B-chain glumate to lysine
B. 6th amino acid substitution of B-chain glumate to valine
C. 121st amino acid substitution of B-chain glumate to glycine
D. 121st amino acid substitution of B-chain glumate to lysine
E. 26th amino acid substitution of B-chain glumate to lysine

Poikilocyte that is present in the peripheral blood in cases of thalassemia.
A. Leptocyte
C. Drepanocyte
B. Echinocyte
D. Acanthocyte

Poikilocyte seen in Leach phenotype.
A. Echinocvte
B. Elliptocyte
C. Acanthocyte
D. Dacryocyte

RBCs that have a thin rim of hemoglobin as to degree of hypochromia should be graded as?
A. 1+
B. 2+
C. 3+
D. 4+

A polychromasia grading of 2+.
A. 1%
B. 3%
C. 5%
D. 10%

Aggregates of 5-10 RBCs piled that are stacked together.
A 1+
B. 2+
C. W+
D. 3+

Fine basophilic stippling.
A. Lead poisoning
B. Sideroblastic anemia
C. Porphyria
D. Polychromatophilia

Cells that contain Howell-Jolly bodies, after passing through the spleen will appear as?
A. Dacryocytes
C.Echinocytes
B. Bite cells
D. Spur cells

Elements that can only be seen if demonstrated with supravital stain.
I. Heinz bodies
II. (y4)
III. (B4)
IV. Reticulocytes

A. I. II. III. IV
B. I, II, III
C. I, III, IV
D. I. II. IV

Presence of IgM in the patient’s blood.
A. Increased RBC, HCT, MCHC
B. Decreased RBC, Increased MCV and MCHC
C. Increased RBC, Decreased MCV and MCHC
D. Decreased, RBC, MCV, MCHC

What is the most probable cause of a decreased BC and HCT in automated hematology analyzers.
A. Icteric, lipemic or turbid sample
B. Hemolyzed sample
C. Nucleated RBCs
D. Increased WBC

Leukocytes have large granules that are peroxidase positive.
A CGD
B. May-hegglin
C. Chediak-Higashi
D. Pelger-Huet

Macrophages appear foamy.
A. Gaucher Disease
B. Niemann-Pick Disease
C. Fabry’s disease
D. Krabbe disease

Most useful for detecting lymphoblasts.
A. MPO
B. PAS
C. TRAP
D. TDT

Also known as Di Guglielmo syndrome.
A. FAB M3
B. FAB M4
C. FAB M5
D. FAB M6

Also known as Schilling leukemia.
A. FAB M3
B. FAB M4
C. FAB M5
D. FAB M6

Pathognomonic for Hodgkin’s lymphoma
A. Sezary cells
B. Hairy cells
C. Reed-Sternberg cells
D. Downey cells

Values that are derived from the histogram.
RDW
II. PDW
III. MCV
IV. PCV

A. I, II, III, IV
B. I, II, III
C. I, II, IV
D. I, III, IV

The marker for human hematopoietic stem cells
A. CD 13
B. CD 34
C. CD 8
D. CD 10

A platelet estimate of 401,000-599.000/ uL.
A. Slightly increased
B. Normal
C. Moderate increased
D. Markedly increased

The primary cause of death in patients with sickle cell anemia
A. Bleeding
C. Infectious crises
B. Anemia
D. Viral infections

The instrument used for electromechanical detection of fibrin clot.
A. Electra 750
C. Fibrometer
B. Fibrin timer series
D. FP 910 Analyzer

Photo-optical detection of fibrin clot formation using semi-automated methods.
A. Ortho Koagulab 16S and 40A
C. Coag-A-Mate X2 and XC
B. Coag-A-Mate X2
D. Electra 750 and 750A

Photo-optical detection of fibrin clot formation using automated methods.
A. Electra 750 and 750A
C. FP910 Coagulation Analyzer
B. Fibrin timer series
D. Ortho Koagulab 16S and 40A

A source of a positive or a negative error.
A. Aperture plugs
B. Extraneous electrical pulses
C. Improper aperture current
D. Excessive lysis

Side angle light scatter.
A. Cell size
B. Cell count
C. Cell granularity and lobularity
D. Cell type

Which of the following red cell inclusions stain with Perl’s Prussian Blue?
A. Howell-Jolly Bodies
C. Pappenheimer bodies
B. Basophilic stippling
D. Heinz bodies

Gauge of the needle indicating the largest bore is:

A. 16 gauge
B. 19 gauge
C. 21 gauge
D. 23 gauge

What is the anticoagulant of choice for the osmotic fragility test?

A. Heparin
B. Double oxalate
C. EDTA
D. Potassium Oxalate

Hemoglobin migration pattern on cellulose acetate from point of application to anode is:

A. C < F < A2 < A < S
B. C < S < A and A2 < F
C. C and A2 < S < F < A
D. C < A < F < S < A2

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