PBS AND DIFFERENTIAL COUNT Flashcards
(121 cards)
1
Q
Materials used:-
Glass slide
-Spreader
-Blood sample (whole blood)
A
WEDGE BLOOD SMEARS
2
Q
-Place a drop of blood 1cm away from labelled end
A
3
Q
-When using blood from finger or heel, prevent touching the skin with the slide
A
4
Q
-Hold the slide with blood in one hand and the spreader on the other hand
A
5
Q
-Place the spreader slightly in front of the blood drop (30-40 degree angle)
A
6
Q
-Draw the spreader slide back
A
7
Q
-As soon as blood comes in contact with spreader, it begins to spread to the edges, if not, wiggle it a little
A
8
Q
-Push the spreader rapidly over the entire length of the slide
A
9
Q
Let dry, stain.
A
10
Q
a) Glass slides must be clean
A
WEDGE BLOOD SMEARS
11
Q
b) As soon as blood is placed on the slide, immediately make the smear
A
WEDGE BLOOD SMEARS
12
Q
WEDGE BLOOD SMEARS delay
A
abnormal distribution of wbc (many large cells accumulate at the thin edge) rouleaux of red cells and plt clumping.
13
Q
WEDGE BLOOD SMEARS
Causes of poor blood smear:
A
-drop of blood too large or too small-Spreader slide pushed in a jerky manner
-Failure to put the entire edge of the spreader slide against the slide while making the smear
-Failure to put the spreader in a proper angle against the slide.
-Failure to push the spreader slide completely across the slide
14
Q
results in thicker smear for very low hct
A
Increase angle
15
Q
gives a thin smear for patients with very high hematocrit
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Decrease angle
16
Q
Too much Drop of blood
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17
Q
Too little Drop of blood
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18
Q
Angle Too high
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19
Q
Angle Too low
A
20
Q
Pressure Decreased
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21
Q
Pressure Increased
A
22
Q
Speed Too slow
A
23
Q
Speed Too fast
A
24
Q
A. Materials used:
-2 cover slips (22mm thick, 0.13-0.17 mm thick)
-Blood sample
A
COVER GLASS SMEARS
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COVER GLASS SMEARS
B. Procedure:
-Hold 1 cover glass by its [?], place a drop of blood
2 adjacent corners
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COVER GLASS SMEARS
B. Procedure:
-With the second hand, hold the [?] as the first
second cover slip
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COVER GLASS SMEARS
B. Procedure:
-Gently place the [?] on top of the one containing blood
second cover glass
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COVER GLASS SMEARS
B. Procedure:
drop of blood in the middle of the 2 glass slides forming
16 sides
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COVER GLASS SMEARS
B. Procedure:
-Just before spreading of blood is complete, separate the 2 cover glasses by a [?] (avoid squeezing the 2 glass slides together)
rapid, even, horizontal, lateral pull
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COVER GLASS SMEARS
B. Procedure:
-Allow the smears to [?]
-Stain
air dry completely
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Cover glasses must be clean
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When obtaining blood from finger tip puncture, do not touch the skin
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The cover glasses should be put together without delay upon dropping of blood
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COVER GLASS SMEARS delay
clumping of plts and wbcs, rouleaux formation
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-Too large drop of blood causes too thick smear
COVER GLASS SMEARS
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-This type of smear prep gives a more even distribution of wbcs than the wedge
COVER GLASS SMEARS
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More time consuming, tedious, harder to master
COVER GLASS SMEARS
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Harder to label
COVER GLASS SMEARS
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Coverslips are easily broken
COVER GLASS SMEARS
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A. Materials used
-Blood sample
-Instrument: Hemaspinner
AUTOMATED SPUN SMEAR
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AUTOMATED SPUN SMEAR
B. Procedure
-A clean glass slide is placed on a plate, [?] are placed at the center of the slide.
3-4 drops of blood
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AUTOMATED SPUN SMEAR
B. Procedure
-When the top of the instrument is closed, the plate spins at a high speed for a period of time (excess blood is thrown from the slide to a basin)
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AUTOMATED SPUN SMEAR
B. Procedure
-Resultant slide is completely covered with a [?]
-Stain
monolayer of cells
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A. Materials used:
-Wintrobe tube
-Capillary pipette, steel needle
-Glass slides-Coverslip
BUFFY COAT SMEAR
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BUFFY COAT SMEAR
B. Procedure:
-Using the capillary pipette, fill a [?] with well mixed whole blood
wintrobe tube
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BUFFY COAT SMEAR
B. Procedure:
-Centri:
15 minutes, 1500g
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BUFFY COAT SMEAR
B. Procedure:
Locate the buffy coat, separate [?] except a small amount near the buffy coat
plasma
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BUFFY COAT SMEAR
B. Procedure:
-Using capillary pipette, remove the
small amount of remaining plasma, the entire buffy coat, small amount of red cells
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BUFFY COAT SMEAR
B. Procedure:
-Place on a glass slide, [?] (do not spread out too much on the slide)
mix
50
BUFFY COAT SMEAR
B. Procedure:
-Transfer a small amount of blood to each of two slides, immediately prepare
wedge or cover and a glass smear
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BUFFY COAT SMEAR
The amount of [?] should be approximately equal to the volume of the cellular portion of the mixture
plasma mixed with the buffy coat and red cells
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Microhematocrit tubes may also be used to centrifuge the specimen
BUFFY COAT SMEAR
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cut the tube [?], transfer blood onto slide
near the buffy coat layer
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[?] on a buffy coat smear does not always make an accurate differential count
Distribution of WBCs
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tend to settle according to their cell type during the centrifugation process
nucleated cells
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the differentials from a well made buffy coat smear correlate relatively well with the standard blood smear
leukopenia
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A. Materials used:
-Blood sample
-Glasss slide
THICK BLOOD SMEAR
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THICK BLOOD SMEAR
B. Procedure
Place [?] in the center of a glass slide
one large drop of blood
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THICK BLOOD SMEAR
B. Procedure
Using the [?], carefully spread the drop of blood over an area the size of a dime
corner of the second slide
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THICK BLOOD SMEAR
B. Procedure
[?]: place newspaper behind the slide.
Spread the drop od blood until the newspaper print is just visible through the blood.
Thickness
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THICK BLOOD SMEAR
B. Procedure
Allow the film to completely dry before staining (?)
4-2 hrs at room temp, preferably overnight
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THICK BLOOD SMEAR
B. Procedure
-If not completely dry, will [?] during staining. (This type of stain is not fixed prior to staining)
wash off
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Stains used:
Romanowsky Stains
-Wright’s (oxidized methylene blue, eosin azures)
-Wright’s
– Giemsa/ Modified (more delicate staining char)
-Leishman
-Jenner
-May-Gruwald
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(oxidized methylene blue, eosin azures)
-Wright’s
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(more delicate staining char)
-Wright’s– Giemsa/ Modified
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oxidized methylene blue
Eosin B or Eosin Y
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oxidized methylene blue products
Azure B
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Are considered polychromatic (Impart different colors to cells and cellular elements)
Romanowsky Stains
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promotes ionization during which time staining takes place
Buffer
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neg charged, azure B: pos charged
eosin ions
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STAINING
Procedure:
-Place the dried smear level on a [?]
-Fix by flooding with methanol-Flood slides with [?], time for 4 minutes
-Without removing the stain, add equal volume of [?]
-Mix 2 solutions on the slide by gently blowing back and forth over the solutions (a [?] should form, time for 7 minutes)-rinse thoroughly with tap water
-Wipe the [?], air dry
staining rack
Wright-Giemsa stains
phosphate buffer
metallic green sheen
back of the slides
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a) Pink to orange
red blood cells
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b) Pinkish-grey
reticulocytes
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c) Dark purple nuclei
lymphocytes and neutrophils
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d) Lighter nucleus
monocytes
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e) Bright orange granules
eosinophils
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f) Dark blue black granules
basophils
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g) Violet to purple granules
platelet
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h) gray blue with fine reddish granules
Cytoplasm of the monocyte
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has a light pink cytoplasm with lilac granules
Neutrophil
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shows varying shades of blue cytoplasm
Lymphocyte
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cell parts taking acidic dyes appear more pink, parts taking basic dye show pale staining
too acidic
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rbcs appear greyish-blue, white cell uclei stain deeply purple
too alkaline
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equal staining of the film
Staining rack should be level
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causes stain to precipitate on the dried smear
Insufficient washing
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fades the stain
Excessive rinsing
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If restain: flood smear with [?] and rinse to remove the stain BUT, new smear and stain is always advisable.
methanol
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For [?], after staining is done, mount the cover slip blood side down onto a glass slide with mounting medium.
coverglass smears
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Stock solutions of individual stains must be freshly prepared how often?
each week
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Staining buffer solution may only be used for
1 hour
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controls pH
Phosphate buffer
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Performed to determine the relative number of each type of white blood cell present in the blood.
DIFFERENTIAL COUNT
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-Study of red cell, white blood cell and platelet morphology is performed
DIFFERENTIAL COUNT
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Approximation of the number of platelets are made
DIFFERENTIAL COUNT
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-Performed after the cell counts have been completed
DIFFERENTIAL COUNT
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-May be used to double check the wbc count
DIFFERENTIAL COUNT
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tend to show increased concentrations of PMNs, monocytes and large abnormal cells at the edges and tail od the smear
Too thin smears
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causes increased lymphocytes at the center of the smear
Too thin smears
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a. Appendicitis
Neutrophilia
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b. Myelogenous leukemia
Neutrophilia
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c. Bacterial infections
Neutrophilia
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Eosinophilia
a. Allergies, allergic reactions
b. Scarlet fever
c. Parasitic infections
d. Eosinophilic leukemia
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absolute increase in neutrophil count
Neutrophilia
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Lymphocytosis
a. Viral infections
b. Whooping cough
c. Infectious mononucleosis
d. Lymphocytic leukemia
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Monocytosis
a. Brucellosis
b. Tuberculosis
c. Monocytic leukemia
d. SBE
e. Typhoid
f. Rickettsial inf.
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DIFFERENTIAL COUNT PROCEDURE:
a) Place the prepared smear prep on the stage of the microscope
b) Examine under [?]
c) Take note of anything [?]
d) Examine under [?]
LPO
unusual or irregular
HPO
107
DIFFERENTIAL COUNT PROCEDURE:
Examine under LPO
a) Check for [?] of white blood cells
b) Examine the [?]
c) If there is an [?] in this area, the diff count may be inaccurate
d) If there are [?], the body of the smear may show decreased plt count
e) Discard smear and make a new one
even distribution
thin peripheral edge
increased number of white cells
plt clumps
108
Take note of anything unusual or irregular
a) Large abnormal looking cells, rouleaux formation
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Examine under HPO
WBC estimate
(average WBC in 10 fields)
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DIFFERENTIAL COUNT PROCEDURES
a) Cross sectional, crenellation
b) Lonhitudinal
c) Battlement
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[?] each white blood cell seen and [?] in a differential counter until 100 cells have been counted.
Identify; record
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a) If any [?] are seen, record on a separate sheet of pad paper
h) Examine red cell morphology in a [?] where small amount of red cells only slightly overlap
a) Note variations from normal, classify irregularities as [?]
nucleated red cells
thin area
slight, moderate, marked
113
Examine plts for number and morphology
a) Determine approximate number of plts per field [?]
b) plt estimate = [?]
(8-20 plts per field)
average number of plts in 10 OIO fields x 20,000
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Results are reported as
percentage of the diff wbc cell types
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The most preferred method of reporting = absolute wbc count
Absolute # of cells/L = % of cell type in diff x WBC/L
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c) Do not progress too far into the [?] bec morphology is difficult to observe here.
thick area
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d) Do not use the [?] as the red cells appear to have no central pallor area
very thin area
118
When WBC count is below 1.0x10^9/L, it may be difficult to find WBCs on the stained smear
May perform a 50 differential (NOTE it), buffy coat smear may be prepared
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If abnormal distribution of cell types:
200 cell differential may be done, results divided by 2 (NOTE it)
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If the diff count shows immature granulocytic cells
shift to the left = [?]
shift to the right = [?]
leukemia
increased numbers of hypersegmented neutrophils
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Smear shows (?).
The white blood cell count is (?).
The differential count is as follows: [?].
No [?] seen.
Platelets are (?).
Smear is (?), (?).
red cell morphology
Decreased/WTNR/Increased
Neutrophils: %, Lymphocytes: %, Monocytes: %, Eosinophils: %, Basophils: %
atypical cells
decreased, adequate, increased
positive/negative for malarial parasites
Genus, species, stage of development
