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Hematology 1 > (M) Week 11: Peripheral Blood Smear > Flashcards

(M) Week 11: Peripheral Blood Smear Flashcards

1
Q

Familiarize the roles of the peripheral blood smear

A
  1. evaluation of anemia
  2. evaluation of thrombocytopenia / thrombocytosis
  3. Identification of abnormal cells
  4. INfections like malaria, microfilaria
  5. Inclusions like basophilic stippling, Howell-Jolly bodies, Cabot ring
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2
Q

What is the first thing to consider in making PBS

A

how to collect the specimen (what specimen is acceptable)

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3
Q

What anticoagulant tube is used for the sample needed for PBS?

A

EDTA

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4
Q

what does EDTA mean?

A

ethylenediaminetetraacetic acid

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5
Q

how does EDTA prevent coagulation?

A

by chelating the calcium

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6
Q

what form of EDTA is more commonly prepared (preferred)

A

liquid form (easily mixed)

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7
Q

PBS should be prepared within _______ hours of drawing the specimen

A

2-3 hours

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8
Q

What will be found if a PBS is created with a sample that is more than five hours old

A

blood cell artifacts

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9
Q

What are the advantages of EDTA samples

A
  1. different capacities per tube = multiple slides created
  2. Slides need not to be prepared immediately
  3. Prevents platelet clumping
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10
Q

What are the disadvantages of using EDTA sample for PBS

A
  1. occurrence of satellitism
  2. pseudothrombocytopenia (false decrease of platelets)
  3. Pseudoleukocytosis (false increase of WBC due to platelet agglutinates)
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11
Q

what is the other cause of platelet satelliotosis

A

platelet specific auto antibody

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12
Q

T or F

in using EDTA blood samples, monocytes can be seen with vacuoles

A

T

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13
Q

T or F

monocytes visualized with vacuoles will greatly affect the examination of the smear

A

F (will not effect)

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14
Q

What anticoagulant should be used to prevent platelet satelliotosis

A

sodium citrate

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15
Q

what is the ratio for blood to sodium citrate

A

9:1

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16
Q

what is the WBC and platelet count dilution factor?

A

1:1

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17
Q

what method of blood extraction is used when you need to make the film or smear on the patient’s side

A

finger / heel puncture (anticoagulated free blood)

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18
Q

platelet clumping is expected when using what anticoagulant

A

heparinized microcollection tube

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19
Q

When there’s no anticoagulant used, you can only make _____ slides, this is corrected by using _________

A

few
heparinized microcollection tubes

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20
Q

Familiarize!

Blood smear preparation

A
  1. Manual wedge technique / wedge method
  2. Coverslip method / technique
  3. Coverslip and slide method
  4. automated blood smear preparation
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21
Q

most convenient and commonly used method of PBS

A

manual wedge technique

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22
Q

This is a dispenser of blood for wedge method, it is inserted through the rubberstopper of the tube

A

Diff-Safe dispenser

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23
Q

What is the advantage of the Diff-Safe dispenser

A

no need to remove the rubber stopper to obtain a drop of blood

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24
Q

what is the ideal diameter of blood used for PBS

A

2-3 mm

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25
Q

when the drop of blood is too large, what is the effect on the PBS

A

long thick film

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26
Q

when the drop of blood is too thin, what is the expected PBS outcome

A

short thin film

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27
Q

what should the angle be when you are holding the pusher slide

A

30 to 45 deg angle

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28
Q

What happens to the result of your PBS preparation if you push forward too slowly?

A

accentuation of poor leukocyte distribution

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29
Q

after smearing the blood, it is fixed with ______ vol / vol of methanol

A

10% volume

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30
Q

how many slides should be created ?

A

2 or more film / smear

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31
Q

you may save ___ unstained in cases where another one is needed

A

1

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32
Q

familiarize the causes of unacceptable PBS

A
  1. shows a chip / rough edge on the spreader slide
  2. when there’s hesitation in forward motion
  3. the spreader slide is pushed to quickly
  4. too small drop of blood
  5. the drop of blood didn’t spread along the width of the slide
  6. Due to dirt or grease or high lipid level of the patient in their blood
  7. uneven pressure applied on the slide
  8. due to time delay
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33
Q

the film is ___________ to ________________ the length of the slide

A

two-thirds
three-fourths

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34
Q

the film should be _______________ shaped, very slidhtly rounded at the feather edge

A

finger shaped

thumb

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35
Q

this feature of PBS is very important when examining your film/ smear microscopically

A

feathery edge

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36
Q

the ________ drop of blood is picked up and spread along the slide

A

whole

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37
Q

The thickness of the smear is determined by the (enumerate all three)

A
  1. angle of the spreader slide
  2. size of the drop of blood
  3. speed of spreading
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38
Q

the (greater/lesser) the angle, the thicker and shorter the smear

A

greater

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39
Q

If the hematocrit is increased, the angle of the spreader slide should be (increased/decreased)

A

decreased

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40
Q

newborns have (increased/decreased) hematocrit

A

increased

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41
Q

If the hematocrit is decreased, the angle of the spreader slide should be (increased / decreased)

A

increased

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42
Q

what causes irregular spread with ridges and long tail in PBS

A

edge of spreader is dirty / chipped / dusty

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43
Q

what causes holes in film

A

slide is contaminated with fat / grease and air bubbles

or lipidemia

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44
Q

due to delay in fixing the smear or causes methanol contaminated with water

A

cellular degenerative changes

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45
Q

this technique is older, rarely used now, more common for blood marrow aspirate smears

A

coverslip techniue

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46
Q

what materials are needed for the coverslip technique

A

2 coverslips (22x22 mm)
blood sample

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47
Q

This technique produces an excellent leukocyte distribution, more accurate for differential counting

A

coverslip technique

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48
Q

what is the disadvantages of the coverslip technique

A

coverslips are too small (hard to label, transpo, stain, and store)

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49
Q

other name for the coverslip method when used for bone marrow preparation

A

crush preparation

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50
Q

how will too much pressure affect the performance of the coverslip method?

A

the film will cause cell rupture

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51
Q

what other material may be used for coverslip technique?

A

slides

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52
Q

this technique may cause an artifactual increase in lymphocyte counts

A

wedge smear

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53
Q

greater trauma in cells leads to smudge or _____________ cell formation

A

basket

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54
Q

what is the more common technique used in the lab (wedge smear / coverslip method)

A

wedge smear

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55
Q

These are the sample machines that we can use for the so called automated preparation of the film

A

automated wedge film maker machines

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56
Q

An advanced automated machine wherein both the
staining and making of the smear are done

A

Coulter LH slide maker and stainer

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57
Q

staining and making of the film can be done every how many seconds?

A

30

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58
Q

what is the basis of automated machines for making a film?

A

hematocrit level

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59
Q

type of automated machine that can only do slide preparation (not advanced)

A

automatic wedge film maker

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60
Q

two ways of drying the blood smear

A

small fan
natural air drying

61
Q

why should blood smears be dried as quickly as possible

A

to avoid drying artifact

62
Q

why is blowing breath on the slide counterproductive?

A

it produces echinocytic RBCs and drying artifacts

63
Q

What causes drying artifacts?

A

humidity
anemic paitents
water absorebd from the air

64
Q

how to avoid artifacts?

A
  1. dry as quickly as possible
  2. keep the stopper of the stain
  3. Fix the slight using pure anhydrous methanol
65
Q

The following are under what type of staining?

 Wright’s & Giemsa
 Leishman
 May Grunwal Giemsa
 Jenner-Giemsa
 Field’s stain

A

Romanowsky staining

66
Q

what is the common denominator of romanowsky stains?

A

eosin Y and azure B-methylene blue composition

67
Q

Romanowsky stains are considered as what type of stain due to composition?

A

polychrome stain

68
Q

What is the routinely used stain?

A

Wright’s & Giemsa stain

69
Q

What stain, when alone, is used for blood parasite detection?

A

giemsa

70
Q

Characteristic of Eosin Y

A

acidic stain

71
Q

What does Eosin Y stain?

A

hemoglobin and eosinophilic granules.

(ACIDIC STAIN)

72
Q

What is the characteristic of Azure B-Methylene blue

A

basic stain such as RNA

73
Q

What reagent is used for fixing PBS stains

A

pure methyl alcohol or methyl alcohol

74
Q

what buffer is used for PBS

A

0.05 M sodium phosphate
aged distilled water

75
Q

what is the pH of sodium phosphate

A

6.4

76
Q

what is the pH of aged distilled water

A

6.4 to 6.8

77
Q

why is a buffer solution needed for PBS

A

staining reactions are pH dependent

78
Q

what indicates proper mixing of eosin Y and methylene blue?

A

metallic sheen

79
Q

how many minutes are the stains left for?

A

1-3 minutes

80
Q

State the Disadvantages and Advantages of Manual Wright Staining of Slides

A

Advantage: Produces a DESIRABLE STAINING QUALITY
Disadvantage: INCREASED RISK OF SPILLING THE STAIN and LONGER TIME IS REQUIRED FOR THE STAINING PROCEDURE (time consuming)

81
Q

TOF. After 3 minutes, wash the back of the smear with TAP WATER

A

T

82
Q

TOF. wipe the back of the slide to remove stain
residues and air dry in a HORIZONTAL position.

A

vertical

83
Q
  • useful for high volume of sample
  • 5 – 10 mins you are done with the staining which is done by batch
  • processes of fixing/staining and buffering are similar in practice to those of the manual method
A

AUTOMATED SLIDE STAINER

84
Q

Disadvantage of AUTOMATED SLIDE STAINER

A

o YOU CANNOT ADD ADDITIONAL SLIDES
once the staining process has began.
o AQUEOUS SOLUTION should be MADE OFTEN.

85
Q

 Another method of staining aside from MANUAL
and AUTOMATED STAINING.
 It is FAST AND EASY, you ONLY NEED 1
MINUTE and then you’re done.

A

QUICK STAINING

86
Q

Used stain for Quick Staining

A

MODIFIED WRIGHT OR WRIGHT-GIEMSA is used as the stain with AGED DISTILLED WATER as the buffer

87
Q

Disadvantage of quick staining

A

THE QUALITY OF THE STAIN
o Needs to be CHANGED every now and
then

88
Q

Features of a Well-Stained Smear: Macroscopically

color

A

PINK TO PURPLE in color

89
Q

Features of a Well-Stained Smear: Microscopically
1. RBC
2. WBC Nuclei
3. Neutrophil’s cytoplasm
4. Eosinophils

state bawat colors

A
  1. Orange - salmon pink
  2. Purple - blue
  3. Pink - tan w/ violet or lilac granules
  4. Bright orange refractile granules
90
Q

What do you do if this macrosopic and microscopic features are NOT SEEN after air drying and staining and if the smear is poorly prepared

A

MAKE A NEW ONE

91
Q

how WBC’s should look like in a well-stained smear:
1. Neutrophil
2. Eosinophil
3. Basophil
4. Lymphocytes
5. Monocyte

A
  1. Pink to tan
  2. Bright orange refractile granules in its cytoplasm
  3. Nuclei should be purple of blue in color
  4. Small and large; reactive lymphocytes
  5. kidney-shaped granules
92
Q

Size of the central pallor

in dia

A

1/3 OF THE WHOLE DIAMETER

93
Q

Water contamination could also result to a?

A

HEAVILY DEMARCATED CENTRAL PALLOR.

94
Q

The laboratory technician is consistently creating a bullet shaped blood film, to correct this he should:
A. Increase the acute angle of the pusher slide
B. Decrease the angle of the pusher slide
C.Push the pusher slide in a smooth and constant pressure
D. Allow the blood to spread across the width of the slide

A

D

95
Q

When the blood film is viewed through the microscope, the RBCs appear redder than normal, the neutrophil is barely visible, the eosinophils are bright red orange? What is the most likely cause?
A. The slide was overstained
B. The stain was too alkaline
C. The buffer was too acidic
D. The slide was not rinsed adequately

A

C

96
Q

Ideal angle (module based)

A

30-45 degree

97
Q

Pink to purple

A bluer overall is caused by what (2)

A
  1. increased protein level (multiple myeloma patients)
  2. rouleaux formation
98
Q

Pink to purple

  • Due to RBC AGGLUTINATION
  • It is when RBCs agglutinate with each other and seen in patients suffering from COLD AGGLUTININ DISEASE.
  • Patients HAVE ANTIBODIES that could agglutinate RBCs
A

Grainy appearance

99
Q

Pink to purple

Holes over the film Indicates?

A

INCREASED LIPID LEVEL and
maybe the patient has LIPEMIA.

100
Q

Pink to purple

Blue specks out of feathery edge Indicates

A

MARKEDLY INCREASED WBC and PLATELET COUNTS.

101
Q

What part of the microscope should be almost all the way up and adjusted correctly for the magnification used.

A

Condenser

102
Q

What part of the microscope should be opened to allow a comfortable amount of light to the eye

A

iris diaphragm

103
Q

What filter should be used to create a whiter light from a tungsten light source.

A

neutral density filter

104
Q

conditions of Koehler illumination

A
  • illuminator should be centered
  • condenser all the way up
  • iris diaphragm should be opened to allow a comfortable amount of light to the eye
  • neutral density filter
105
Q

MICROSCOPIC MAGNIFICATION

  • Assess overall film quality, color and cell distribution
  • Feathery Edge and Lateral Etches (WBC distribution)
  • RBC agglutination and Roulex formation
A

10x Objective Examination (LPO)

106
Q

MICROSCOPIC MAGNIFICATION

Select an assessment area to begin diff count and evaluate cell morphology
WBC estimate (2000x multiplication factor)

A

40x Objective Examination
(HPO)

107
Q

MICROSCOPIC MAGNIFICATION

Assess
WBC Diff count
RBC, WBC and platelet morphology
Platelet estimate
RBC (howell-jolly bodies) and WBC (Dohle bodies) inclusion
Reactive or abnormal cells
Nucleated RBCs - reported per 100 WBCs

A

100x Oil Immersion Objective Examination (highest)
1000x

108
Q

MICROSCOPIC MAGNIFICATION

Optimal assessment area: best possible area for peripheral blood film examination; look for feathery edge (depending on the organism)
WBC Diff. count and morphology exam; WBC estimate (3000x)
RBC, WBC and platelets is examined

A

50x Oil Immersion Objective Examination

109
Q

abnormalities in the size of our RBC.

A

ANISOCYTOSIS

110
Q

feature of most anemias.

A

Anisocytosis

111
Q

(micro, normo or macrocytic) red cells are commonly
associated with certain anemia such as:
 Iron deficiency
 Anemia
 Thalassemia
 Sideroblastic anemia
 Lead toxicity
 Anemia of chronic disease

A

Micro

112
Q

(micro, normo or macrocytic) red cells are seen on patients with MEGALOBLASTIC ANEMIA.

A

Macro

113
Q

(micro, normo or macrocytic) red cell is found IN BONE MARROW AND KIDNEY FAILURE.

A

Normo

114
Q

Artifacts from excess EDTA, Improper smear preparation, prolonged sample storage before blood film preparation

A

Echinocytes

115
Q

an increase disproportionately to its volume that results from a decrease in hemoglobin as in IRON DEFICIENCY ANEMIA or an increase in the cell membrane

A

Target cell/ Codocytes

116
Q

Fragmented RBCs/ Schistocytes

A

RBC injury from damaged endothelium, a feature of microangiopathic hemolytic anemia

117
Q

do not bend or move easily and can block blood flow to the rest of the body

A

Sickle cell RBCs

118
Q

dense, shrunken, irregularly shaped RBCs w/ spikes on the outside, forms from changes in fats and proteins on RBCs outer layers

A

Acanthocytes/Spur cells

119
Q

ENUMARATE ALL RBCs SHAPE

A

 TEARDROP CELL
 BURR CELLS
 DACROCYTE
 ELLIPTOCYTES  STOMATOCYTES  SPHEROCYTES
Echinocytes
Target cell/ Codocytes
Fragmented RBCs/ Schistocytes
Sickle cell RBCs
Acanthocytes/Spur cells:

120
Q

Hemoglobin Conc.

mature RBCs w/ normal hemoglobin

A

Normochromic RBC

121
Q

Hemoglobin Conc.

central pallor more than ⅓ of the total diameter, decreased hemoglobin conc.

A

Hypochromic RBCs

122
Q

Hemoglobin Conc.

central pallor less than ⅓, increased hemoglobin conc., red cells stains deeply

A

Hyperchromic RBC

123
Q

TOF. Sickle cell is reported as positive or negative, RBC grading is not used according to other textbooks.

A

T

124
Q
  • presence of blue-gray to pink cytoplasm tint of the red cell due to either hemoglobin presence or RNA residue.
  • It is commonly SEEN IN YOUNG RED CELL.
  • indicates RETICULOCYTOSIS.
  • RBCs appears LARGER THAN NORMAL AND LACKS CENTRAL PALLOR WITH BLUE-GRAY TO PINK CYTOPLASM.
A

POLYCHROMATOPHILIA

125
Q

what is the disadvantage of leaving the stain for more than 3 minutes

A

increased risk of spilling the stain

126
Q

MATCH

Supravital staining is not limited to reticulocyte it can also determine:
* Heinz bodies
* Howell Jolly bodies
* Hgb H
* Pappenheimer bodies

A. presence of DNA content.
B. presence of hemoglobin H in your RBCs.
C. denatured hemoglobin
D. indicates the presence of iron (hemosiderin)

A

CABD

127
Q

Which among these is not graded as +1, +2,
+32
A. Sickle cell
B. Pappenheimer bodies
C. Howell jolly bodies
D. Basophilic stipplings
E. A,B,D
F. A, B,C,D

A

F

128
Q
  • when cells aggregate into cluster of masses when exposed to an anti-body;
  • happened because of the presence of antibodies that agglutinates the RBC
A

Agglutination

129
Q

 coin or stacking appearance of RBC;  common in increased level of RBC

A

Rouleau Formation

130
Q

what should the drying position be?

A

vertical position

131
Q

 Size -1-3μm
 Non-nucleated cells derived from cytoplasmic
fragments of Megakaryocytes  Has purple red granules.
 Lilac color

A

Platelets

132
Q

machine used for staining, done by batch, it’s useful for a high volume of sample

A

automated slide stainer

133
Q

 Clinically significant
 Indicative of certain condition  same in size of the RBC

A

Giant Platelets

134
Q

what is the disadvantage of Hema-Tek 2000 slide stainer

A
  1. you cannot add additional slides one the staining process has begun
  2. aqueous solution should be made often
135
Q

WBC count formula 1 (40x magnification)

A

Ave. WBC per field x 2000

136
Q

this is another method of staining aside from manual and automated staining

it is fast and easy, you only need one minute to be done

A

quick staining

137
Q

WBC count formula 2 (x50 magni)

A

Ave. WBC per field x 3000

138
Q

Formula 1 and 2 for platelt count

A

 Formula 1: Average platelet per field x 20,000
Formula 2: Average # of platelet per fields x RBC ct./ 200 RBC

139
Q

what stains may be used for quick staining

A

modified wright
modified giemsa

140
Q

what is the buffer used in quick staining

A

aged distilled water

141
Q

what are the disadvantages of quick staining

A

quality of the stain
needs to be changed every now and then

142
Q

EXAMINATION OF BLOOD FILMS FOR PARASITES

when parasites are scanty

A

Thick film

143
Q

what should the color of stained blood be?

A

pink to purple (macroscopically)

144
Q

EXAMINATION OF BLOOD FILMS FOR PARASITES

identification of species

A

Thin film

145
Q

EXAMINATION OF BLOOD FILMS FOR PARASITES

Staining of Film by?

A

Giemsa or Leishman’s stain at pH 7.2

146
Q

what should be the color of WBC nuclei in a well stained smear

A

purple to blee

147
Q
A
148
Q

what is the ideal color of neutrophil’s cytoplasm in a well stained smear

A

pink-tan with violet / lilac granules

149
Q
A

Hematology 1 (32 decks)

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