HEMA 1 Flashcards
The average human possesses ______
5L of blood
5-6L -total blood volume
Composition of blood
Plasma 55%
Formed elements 45%
4 layers of centrifuged tube
- FATTY
- PLASMA
- BUFFY
- RBC’s
Treat all specimens such as blood, body fluids and unfixed tissues to be potentially infectious
STANDARD PRECAUTION
most effective way of breaking the chain of infection
HANDWASHING
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
The process of destroying PATHOGENIC microorganisms in inanimate objects
DISINFECTION
Destruction of all forms of microbial life including spores
Sterilization
An appropriate disinfectant is a household bleach (_________), used in a _________.
sodium hypochlorite
1:10 v/v
The process of obtaining blood from a vein
VENIPUNCTURE
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)
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
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.)
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
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
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
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
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
BLACK
Anticoagulant/additive:
Specimen type/use:
Mechanism of action:
Anticoagulant/additive: Na citrate
Specimen type/use: Plasma/ ESR
Mechanism of action: Chelates/ binds calcium
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
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
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
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
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
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
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
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
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
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
EDTA containing tubes
Lavender
Pink
White
Royal blue
Tan (plastic)
Na Citrate containing tubes
Light blue
Black
Aka SEQUESTRENE
K3 EDTA
Aka versene
K2 EDTA
Aka versene
Na2 EDTA
Short Draw——> OVERANTICOAGULATION —->shrinking RBC can result to:
False ⬇️ HCT & ESR
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)
Blood to anticoagulant ratio using black tube
4:1
Inhibits thrombin
Enhances anti-thrombin-III
Causes BLUE background in PBS
HEPARIN
Inhibits thrombin
Enhances anti-thrombin-III
Causes blue background in PBS
HEPARIN
Contains thrombin
For STAT chem
An additive
ORANGE
Antiglycolytic
Inhibits enolase by binding magnesium
Also inhibits urease
NaF
Substitutes NaF if both glucose and BUN are requested
Li Iodoacetate
Preferred concentration of OXALATE
1-2 mg/mL
Preferred concentration of CITRATE
3.2 g/dL (0.105 M)
3.8 g/dL (0.129 M)
Preferred concentration of EDTA
1-2 mg/mL (chem)
1-5 mg/mL (Hema)
Preferred concentration of FLUORIDE
10 mg/mL
Preferred concentration of HEPARIN
0.2 mg/mL
8x inversions
- YELLOW (SPS, ACD)
- ORANGE (THROMBIN)
- GREEN (HEPARIN)
- LAVENDER AND PINK (EDTA)
- GRAY
5x inversions
- RED (PLASTIC)
- GOLD
3-4x inversions
LIGHT BLUE
Tourniquet- should be applied_________above the venipuncture site and left no longer than ______.
3-4 (7.5-10 cm)
1 minute
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
Most common means of collecting blood specimen is through the use of an evacuated tube system.
ETS
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
Prevents blood from clotting (E.g. EDTA, Potassium oxalate)
Anticoagulant
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
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
Routinely used gauge needles
19-, 20- and 21-gauge needles
Needle guage used in pediatric
23-25 gauge
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).
Most common skin cleanser
70% isoprophyl alcohol
Used for sample collection for blood measurements.
Benzalkonium chloride or nonalcoholic anti-septic
most preferred vein
largest, closest to the surface and well anchored
MEDIAN CUBITAL
2nd option, less anchored, however it easiest vein to palpate in obese patients.
CEPHALIC VEIN
least anchored
close to median cutaneous nerve and brachial artery
BASILIC VEIN
Skin puncture is performed in:
- Newborns, pedia below 1y/o
- Adults who are severely burned
- Elderly patients
Capillary blood values as compared to venous blood:
⬇️rbc ct.
⬇️hct.
⬇️hb.
⬇️plt. ct.
⬆️glucose and WBC ct.
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
capillary blood collection sites
- Lateral side of the plantar surface of the heel- children
- Third or fourth finger- older children or adult
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
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
Punctures should not be more than
________ because of the risk of bone injury or possible infection (osteomyelitis)
2 mm
Cortisol, ACTH, Fe ______
Eosinophils ________
Cortisol, ACTH, Fe- ⬆️ in AM
Eosinophils- ⬆️ in PM/evening
During stress, WBC ct. and acid base balance ________.
INCREASED
During exercise, creatinine, protein, CK, AST, LD, platelet and WBC ct., HDL _______
INCREASE
Prolong standing ________.
Matagal na nakahiga_________.
HEMOCONCENTRATION
HEMODILATION
In smoking WBC count _______
INCREASE.
CBC
- WBC count
- RBC count
- Hb
- Hct.
- WBC Differential
- RBC indices
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
Hemoglobin ____ in the morning
Increased
Modified Drabkins Reagents
Potassium cyanide
Potassium ferricyanide
Donates cyanide to hemoglobin
Potassium cyanide
Converts Fe2+ to Fe3+
Potassium ferricyanide
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)
decreases amount of turbidity resulting from abnormal proteins and improves RBC lysis
Nonionic detergent
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
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.
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
Hemoglobin S and C
Result:
Remedy:
Result: turbidity and falsely high results
Remedy: dilute mixture 1:2 with water then multiply results by 2
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
•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)
Blood iron content
100g of Hb=_______ of Fe2+
100g of Hb= 0.347 g
•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
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
- 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
- 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
- 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
• ________\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
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
Contains heparin (anticoagulated tube), to be used for samples that are non-anticoagulated.
RED BAND
Plain tubes (non-anticoagulated tube), to be used for samples that are anticoagulated.
Blue band
Clay like sealing compound
4-6 mm
Microhematocrit centrifuge capable of _______ RCF for _______ minutes.
10k-15k RCF
5 minutes
Microhematocrit specimen:
________ whole blood is preferred
K3-EDTA causes a _____ decrease in the hematocrit due to shrinkage of the RBCs
K2-EDTA
2-3%
incomplete sealing (less than 4-6 mm), leads to ________
falsely low results
Inadequate centrifugation (shorter than 5 minutes), leads to a __________.
falsely increased result
Allowing the tube to stand longer than several minutes leads to ______.
falsely increased result
Overanticoagulation can cause hematocrit ______
Falsely low results
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
Increased amount of trapped plasma is found in:
- Macrocytic anemia
- Spherocytosis
- Thlassemia
- Hypochromic anemia
- Sickle cell anemia
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
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
Thoma red count pipet marks and there dilution:
0.5 (1:200)
101 (1:100)
RBC count diluting fluid
1.Gower
2. Eagle
3. NSS
4. TOISON
5. STRONG
6. BETHEL
7. HAYEM
8. DACIE
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
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)
Or
815 x 200/10 (0.004) = 163,000/ 0.04
= 4.075 x 10^12/L
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
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
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
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)
WBC estimation DILUTION FACTOR
2000
Dilution for Normal WBC count
1:10 or 1:20
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)
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)
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)
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
RBC
Diluent:
Dilution:
Objective:
Area:
Diluent: ISOTONIC SALINE
Dilution: 1:100
Objective: 40x
Area: 0.2 mm^2
Platelets
Diluent:
Dilution:
Objective:
Area:
Diluent: 1% ammonium oxalate
Dilution: 1:100
Objective: 40x, phase
Area: 1 mm2
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
Size: 10-15 um
Nucleus: elongated, curved or sausage shaped.
Cytoplasm: Identical to segmented neutrophil.
Other name: Nonsegmented neutrophil, neutrophil staff or stab
BAND
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
Size: 10-14 um
Nucleus: Light to purple staining, usually difficult to see due to overlying granules
Cytoplasm: Densely stained, dark violet granules
BASOPHIL
All of the following shift to the left, except:
A. Myelocyte
B. Metamyelocyte
C. Band (if >6)
D. Segmenters
D. Segmenters
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
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
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
A shift to the right refers to an ________
increased number of hyper segmented neutrophils
Granulocytes
Neutrophil
Eosinophil
Basophil
Non-granulocytes
Monocytes
Lymphocytes
Polymorphonuclear
N
E
B
Mononuclear
M
L
Phagocytes
N
E
B
M
Immunocyte
Lymphocyte
NEUTROPHILIA, except:
a. Appendicitis
b. Allergies
c. Myelogenous leukemia
d. Bacterial infection
B. Allergies
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
EOSINOPHILIA, except:
a. Parasitic infections
b. Scarlet fever
c. Allergies
d. Immunodificiency
D
Eosinopenia, except:
a. Brucellosis
b. Decreased production
c. Acute bacterial infection
d. ACTH administration
A
Brucellosis
Tuberculosis
Subacute Bacterial Endocarditis
Typhoid
Rickettsial infections
Hodgkin’s disease
Gaucher disease
MONOCYTOSIS
Glucocorticoids
Overwhelming infections that also cause neutropenia
MONOCYTOPENIA
Viral infections
Whooping cough
Infectious mononucleosis IM
Lymphocytic leukemia
LYMPHOCYTOSIS
Stress
Hyperthyroidism
Increased glucocorticoid levels
BASOPENIA
Long-term drug therapy
Immunodeficiency
LYMPHOCYTOPENIA
Immediate hypersensitivity reactions
Hypothyroidism
BASOPHILIA
Indicates the average volume of RBCs in femtoliters (fL)
MCV
MCV FORMULA
NV
hct/rbc ct. x 10
80-100 fL
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
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
Determined from the RBC histogram; coefficient of variation of the MCV
RDW
RR: 11.5-14.5%
⬆️ 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)
MICROCYTIC, HYPOCHROMIC
ATIS
Anemia of chronic inflammation
Thalassemia
IDA
Sideroblastic anemia
NORMOCYTIC, NORMOCHROMIC
AHA
1. Aplastic anemia
2. Hemolytic anemia
3. Acute blood loss anemia
MACROCYTIC, NORMOCHROMIC
LMM
1. Liver disease
2. Myelodysplasias
3. Megaloblastic anemia
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 °
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.
FACTORS THAT AFFECT THE THICKNESS OF A WEDGE SMEAR
Pressure
Angle
Size of blood drop
Speed
Thick smear
P ⬇️
A ⬆️
S ⬆️
S ⬆️
Thin smear
P ⬆️
A ⬇️
S ⬇️
S ⬇️
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)
EXCESSIVELY BLUE STAIN
Thick films
Prolonged staining time
Inadequate washing
Too high alkalinity of buffer
EXCESSIVELY PINK STAIN
Insufficient staining time
Prolonged washing time
Mounting the coverslip before the slide is dry
High acidity of the stain
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
Important in helping to diagnose bleeding disorders.
Platelet count
NV: 150,000-450,000/ uL (150-450 x 10^9/L)
THROMBOCYTOSIS
PICS
1. Polycythemia vera
2. Idiopathic thrombocythemia
3. Chronic myelogenous leukemia (CML)
4. Splenectomy
THROMBOCYTOPENIA
TAAG PS
1. Thrombocytopenia purpura
2. Aplastic anemia
3. Acute leukemia
4. Gaucher’s disease
5. Pernicious
6. Splenomegaly
MPV should be less than ____ hours
4
If >4 hours old MPV will have _____ increment.
20%
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
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
Place the charged hemacytometer in a moist chamber for ________ to allow the platelets to settle.
15 minutes
____________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
Retic count formula and normal values.
%RC=# of retics per 1000 RBC’s/ 10
NV: 0.5-1.5% (adult)
2-6% (newborn)
⬇️ retic count
- aplastic anemia
- conditions in which the bone marrow is not producing red blood cells
⬆️ retic count
- hemolytic anemias
- individuals with IDA receiving iron therapy
- thalassemia
- sideroblastic anemia
- acute and chronic blood loss anemia
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
Spleen- saquesters
Sequest 20-30% of platelets
Retic count sources of error: 3 H
Heinz body
HbH
Howell jolly
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
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
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
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
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
Rouleaux formation
Agglutination
Macrocytes
Severe anemia
INCREASED ESR
Sickle cells and spherocytes
Anisocytosis
Poikilocytosis
DECREASED ESR
The single most important factor in determining ESR
PLASMA COMPOSITION
ESR = RBC MASS
ESR ∝ PLASMA VISCOSITY
ex. Increased albumin = decreased ESR
A tilt of 3° can cause errors up to ______
30%
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
vibrations from opening and closing the refrigerator doors
Falsely INCREASED ESR
heat released from the refrigerator motor
Falsely INCREASED ESR
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
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
Hypercholesterolemia
Hyperfibrinogenemia
Hypergammaglobulinemia
Hypoalbuminemia
INCREASED ESR
Multiple myeloma
Rheumatoid arthritis
INCREASED ESR
Acanthocytosis
Anisocytosis (marked)
Hemoglobin C
J. Microcytosis
Polycythemia
Sickle cells
Spherocvtosis
Thalassemia
DECREASED ESR
Anemia
Macrocytosis
INCREASED ESR
Refrigerated sample not
returned to room temperature
INCREASED ESR
- Clotted blood sample
- Delay in testing
DECREASED ESR
Bubbles in ESR column
Low room temperature
Narrow ESR column diameter
DECREASED ESR
High room temperature
Tilted ESR tube
Vibration
INCREASED ESR
LEUKOCYTOSIS
DECREASED ESR
LEUKEMIA
INCREASED ESR
Hyperalbuminemia
Hyperglycemia
Hypofibrinogenemia
Hypogammaglobulinemia
DECREASED ESR
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
2x
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
G1
RNA and PROTEIN synthesis
10 hrs
Synthesis (S) phase
DNA synthesis
9 hours
G2
Premitotic stage
4 hrs
Mitosis
Cell division
IPPAT
1 hour
G0
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
C
- 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
D
- 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
C
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
C
- 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
B
- 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
A
- 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
B
- 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
D
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
C
- 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
B
- 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
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
C
Diluting fluids used in RBC count EXCEPT:
A. Dacie
B. Eagle
C. Bethel
D. Turk’s
D
- A neutrophil that has a curved or sausage-shaped nucleus
A. Segmenter
B. Band
C. Juvenile
D. PMN
B
- The youngest WBC that is normally present in the differential count.
A. Segmenter
B. Juvenile
C. Neutrophil metamyelocyte
D. Band
D
- The first stage that allows granulocte differentiation.
A. Segmenter
B. Promyelocyte
C. Myelocte
D. Metamyelocyte
C
- 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
B
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
C
- The anticoagulant of choice for OFT.
A. Heparin
B. Citrate
C. EDTA
D. Oxalate
A
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
C
- 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
C
- 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
C
The manual reticulocyte count values should agree within?
A. 5%
B. 10%
C. 15%
D. 20%
D
- 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
C
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
B
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
A
- 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
C
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
A
- 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
C
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
C
- Which cytokine negatively affects cell proliferation?
A. IL- 1
B. TGF-B
C. IL-9
D. IL- 11
B
- 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
A
- Dendritic cells are believed to arise from?
A. Common lymphoid progenitor
B. Monocytes
C. Lymphocytes
D. T-cells
B
- A stain that is used to differentiate FAB M6 (acute erythroleukemia) from ALLs.
A. Toluidine blue
B. PAS
C. TDT
D. TRAP
B
Tiny B lymphocytes that have fine cytoplasmic extensions possess.
A. Myeloperoxidase
B. Esterases
C. ACP 5
D. LAP
C
A WBC that has ALP.
A. Neutrophil
B. Monocyte
C. Lymphocyte
D. Eosinophil
A
Which of the following WBCs will be negative for esterase?
A. Neutrophil
B. Monocyte
C. Lymphocyte
D. Eosinophil
C
- 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
C
- 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
D
- 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
B
- Tests for PH EXCEPT:
A. Sugar water screening
B. Sucrose hemolysis
C. Sodium metabisulfite
D. Acid serum test
C
- Megakaryocytic stage characterized by an irregularly shaped nucleus.
A. Megakaryoblast
B. Promegakaryocyte
C. Megakaryocytic
D. Metamegakaryocyte
B
- Lifespan: 9-10 days from myeloblast to death.
A. Neutrophil
B. Monocyte
C. Lymphocyte
D. Eosinophil
A
- Maturation 3.5 days, lifespan: 18 hours
A. Neutrophil
B. Monocyte
C. Lymphocyte
D. Eosinophil
D
- Maturation: 30-48 hrs, lifespan: months
A. Neutrophil
B. Monocyte
C. Lymphocyte
D. Eosinophil
B
- Maturation: 1-2 days, lifespan: months to years
A. Neutrophil
B. Monocyte
C. Lymphovte
D. Eosinophil
C
- 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
D
- 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
A
- 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
A
- 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
B
- 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
B
- 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
C
- 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
D
- 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
- A stain that differentiates acute myelogenous leukemia and acute monocytic leukemia from ALL
A. MPO
B. SBB
C. PAS
D. Specific esterase
A
- 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
C
- The most commonly used system for venipuncture.
A. Syringe
B. Evacuated tube
C. Butterfly needle
D. Lancet
B
- 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
D
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
C
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
B
- 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
C
- 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
B
- 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
S
- 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
B
- A gray top tube contains what anticoagulant?
A. Na fluoride
B. lodoacetate
C. Oxalate
D. Citrate
C
- 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
B
- 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
A
- 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
C
- What color stopper must be used for ESR?
A. Blue
B. Black
C. Lavender
D. Red
B
- 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
B
- 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
A
- 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.
C
- 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
C
- 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
D
- 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
C
- It enhances AT-Ill activity in vivo.
A. Heparan sulfate
B. Adenosine
C. ATP
D. Serotonin
A
- The enzyme that is inhibited by aspirin.
A. Lipoxygenase
B. Cyclooxygenase
C. Thromboxane synthetase
D. PGI2 synthetase
B
- The protein which is responsible for maintaining the shape of the platelets.
A. Actin
B. Myosin
C. Actomyosin
D. Tubulin
D
- Which of the following does not refer to HMWK?
A. Flaujeac factor
B. Williams Factor
C. Fletcher factor
D. Fitzgerald factor
C
- Secreted by the dense granules.
A. Platelet cofactor 4
B. ATP
C. Thrombospondin
D. Thromboglobulin
B
- Functions of Factor XIla. EXCEPT:
A. Initiation of fibrinolytic system
B. Conversion of prekallikrein to kallikrein
C. Production of kinins
D. Promotes thrombosis
D
- Which of the following are found in serum?
A. Factor I
B .Factor lI
C. Factor V
D. VIII
E NOTA
E
- 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
B
- 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
B
- A test which uses thromboplastin as its reagent.
A. Prothrombin time
B. АРТТ
C. Plasma recalcification time
D. Lee White Clotting Time
A
- Sensitive for factor VIII Deficiency
A. Prothrombin time
B. APTT
C. Plasma recalcification time
D. Lee White Clotting Time
B
- An INR that is above 3.0 indicates:
A. Increased risk for haemorrhage
B. Increased risk for thrombosis
C. Normal
D. Increased fibrinolysis
A
- 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
B
- 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
C
- It was used to help between factor X and VII deficiencies
A. Reptilase time
C. Duckhert’s test
B. Stypven time
B
- 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
C
- 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
A
- 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
C
- 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
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
D
- 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
A
- Bleeding will not be prolonged in:
A. VWD
B. Bernard-Soulier syndrome
C. Glanzmann thrombasthenia
D. NOTA
D
- 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
B
- A platelet estimate of 50,000-99,000 should be reported as?
A. Markedly decreased
B. Moderately decreased
C. Slightly decreased
D. Low normal
B
- Platelet count is <30,000/ uL.
A. Abnormal BT
B. Bleeding possible with trauma
C. Spontaneous bleeding possible
D. Severe spontaneous bleeding
C
- Normal response to ristocetin, weak response to ADP, epinephrine and collagen.
A. Bernard-soulier
C. Hemophilia A
D. VWD
D. Glanzmann thrombasthenia
D
- Vomiting of blood.
A. Hemoptysis
B. Hematoma
C. Hematemesis
D. Hematuria
C
- Due to a peptidase enzyme deficiency.
A. Pseudoxanthoma elasticum
B. Ehlers-Danlos
C. Scurvy
D. Kassabach-Meritt Syndrome
B
- Relationship of the number of blood units transfused to a patient’s degree of thrombocytopenia.
A. Indirect
B. Direct
C. Unrelated
D. Inverse
B
- Platelet retention in paraproteinemias.
A. Increased
B. Decreased
C. Unaffected
D. Slightly increased
B
- 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
C
- A disease characterized by a triad of recurrent infections, thrombocytopenia and eczema.
A. WAS
C. Hermansky-Pudlak
B. Gray platelet syndrome
D. Chediak-Higashi
A
- 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
A
- 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
B
- Characteristic blood picture of acute blood loss anemia.
A. Microcytic, hypochromic
B. Normocytic. normochromic
C. Macrocytic, hypochromic
D. Macrocytic, normochromic
B
- Which of the following does not produce a microcytic, hypochromic blood picture?
A. IDA
C. Thalassemia
B. ACD
D. Aplastic anemia
D
- All of the following exhibit a normocytic, normochomic blood picture EXCEPT:
A. ACD
B. Lead poisoning
C. Hemolytic anemia
D. Thalassemia
D
- Cells are more susceptible to lysis when exposed to a hypotonic solution.
A. Hereditary elliptocytosis
C. Hereditary spherocytosis
B. Hereditary stomatocytosis
D. Acanthocytosis
C
- Associated with abetalipoproteinemia.
A. Hereditary elliptocytosis
B. Hereditary stomatoctosis
C. Hereditary spherocytosis
D. Acanthocytosis
D
- 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
C
- 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
C
- Which of the following enzymes is only present in neutrophils?
A. Myelperoxidase
C. Chloroacetate esterase
B. ACP
D. LAP
D
- 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
B
- Requires a fresh heparinized sample.
A. LAP
B. MPO
C. SBB
D. Specific esterase
A
- The presence of a biphasic antibody causes hemolvsis in red blood cells
A. PNH
B. PCH
C. Spherocytosis
D. Acanthocvtosis
B
- The poikilocyte that is has decreased osmotic fragility.
A. Spherocytes
B. Acanthocytes
C. Codocytes
D. Dacryocytes
C
- 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
B
- Poikilocyte that is present in the peripheral blood in cases of thalassemia.
A. Leptocyte
C. Drepanocyte
B. Echinocyte
D. Acanthocyte
A
- Poikilocyte seen in Leach phenotype.
A. Echinocvte
B. Elliptocyte
C. Acanthocyte
D. Dacryocyte
B
- 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+
D
- A polychromasia grading of 2+.
A. 1%
B. 3%
C. 5%
D. 10%
C
- Aggregates of 5-10 RBCs piled that are stacked together.
A 1+
B. 2+
C. W+
D. 3+
B
- Fine basophilic stippling.
A. Lead poisoning
B. Sideroblastic anemia
C. Porphyria
D. Polychromatophilia
A
- Cells that contain Howell-Jolly bodies, after passing through the spleen will appear as?
A. Dacryocytes
C.Echinocytes
B. Bite cells
D. Spur cells
B
- 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
C
- 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
B
- 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
B
- Leukocytes have large granules that are peroxidase positive.
A CGD
B. May-hegglin
C. Chediak-Higashi
D. Pelger-Huet
C
- Macrophages appear foamy.
A. Gaucher Disease
B. Niemann-Pick Disease
C. Fabry’s disease
D. Krabbe disease
B
- Most useful for detecting lymphoblasts.
A. MPO
B. PAS
C. TRAP
D. TDT
D
- Also known as Di Guglielmo syndrome.
A. FAB M3
B. FAB M4
C. FAB M5
D. FAB M6
D
- Also known as Schilling leukemia.
A. FAB M3
B. FAB M4
C. FAB M5
D. FAB M6
C
- Pathognomonic for Hodgkin’s lymphoma
A. Sezary cells
B. Hairy cells
C. Reed-Sternberg cells
D. Downey cells
C
- 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
A
- The marker for human hematopoietic stem cells
A. CD 13
B. CD 34
C. CD 8
D. CD 10
B
- A platelet estimate of 401,000-599.000/ uL.
A. Slightly increased
B. Normal
C. Moderate increased
D. Markedly increased
A
- The primary cause of death in patients with sickle cell anemia
A. Bleeding
C. Infectious crises
B. Anemia
D. Viral infections
C
- The instrument used for electromechanical detection of fibrin clot.
A. Electra 750
C. Fibrometer
B. Fibrin timer series
D. FP 910 Analyzer
C
- 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
D
- 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
D
- A source of a positive or a negative error.
A. Aperture plugs
B. Extraneous electrical pulses
C. Improper aperture current
D. Excessive lysis
C
- Side angle light scatter.
A. Cell size
B. Cell count
C. Cell granularity and lobularity
D. Cell type
C
- 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
C
Gauge of the needle indicating the largest bore is:
A. 16 gauge
B. 19 gauge
C. 21 gauge
D. 23 gauge
A
What is the anticoagulant of choice for the osmotic fragility test?
A. Heparin
B. Double oxalate
C. EDTA
D. Potassium Oxalate
A
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
B
