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Hematology > Exam 3 > Flashcards

Exam 3 Flashcards

1
Q

What measurements in the hemacytometer in the large squares and in small squares, what is the total volume on one side, how much depth?

A

large squares, 1x1
small center squares 0.2 x 0.2 mm
0.1 depth
total volume 0.9mm

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2
Q

What is the formula to get result from hemacytometer

A

total count = (cells counted x dilution factor)/ area x depth

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3
Q

What is the dilution in WBC counts

A

1:20

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4
Q

What is the corrected WBC count

A

if there are WBCs in an RBC count, they are indistinguishable,
must use calculation
corrected WBC count= (uncorrected WBCx100)/ RBCs per 100 WBC +100)

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5
Q

What do we use to dilute a plt count, what dilution factor do we use

A

ammonium oxalate, 1:20

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6
Q

What is the cyanmethemoglobin method for hgb

A

drabkin solution
hgb-> oxidized by potassium ferricyanide into methemoglobin -> potassium cyanide turns it into cyanmethemoglobin
cyanomethemoglobin is absorbed at 540nm

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7
Q

What are the common sources of error from a microhematocrit reader

A

sensitive to light
high WBC, plt, lipemia or abnormal hgb give high results
highly toxic

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8
Q

What are the common sources of error from a microhematocrit reader

A

sensitive to light
high WBC, plt, lipemia or abnormal hgb give high results
highly toxic

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9
Q

Define hematocrit

A

volume of packed RBCs in whole blood
packed cell volume

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10
Q

What could cause error in hematocrit test results

A

not sealing the capillary tube correctly
too much anticoagulant shrinks RBCs
not mixed
overcentrifuged
reading results wrong

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11
Q

What is the rule of three in hematology, what if it is not met

A

The value of the hematocrit should be 3x the hgb (+-3)
should be examined for abnormal RBCs could be lipemia, spherocytes, hypochromic and microcytic

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12
Q

Name the RBC indicies

A

MCV- mean cell volume
MCH- mean cell hemoglobin
MCHC- mean cell hemoglobin concentration
RDW- red distribution width

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13
Q

What are the possible indications in RBC morphology for MCV and MCHC

A

MCV microcytic (<80), normocytic (80-100), macrocytic (>100)
MCHC hypochromic (<32) normochromic (32-36) spherocytes (>36)

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14
Q

What is the MCV formula

A

HCT% x 10/ RBC

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15
Q

What is the MCH formula

A

Hgb x 10/ RBC

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16
Q

What is the MCHC

A

HGB x 100/ HCT

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17
Q

Why is a reticulocyte count performed

A

to assess the erythropoietic activity of the BM

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18
Q

What is used to stain the retic count

A

new methylene blue

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19
Q

What is the reticulocyte count calculation

A

retic = # retics x 100/ 1000 RBCs counted

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20
Q

a nonnucleated RBC with 2 or more particles of blue stained granulofilamentous material is considered

A

reticulocyte

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21
Q

What is a miller disk

A

for counting 1000 RBCs, has a large and small square, small square for RBCs large one for retics

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22
Q

What is an ARC?

A

absolute retic count- number of retics in 1L

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23
Q

What is the RPI

A

retic production index
used to correct hematocrit if high retics

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24
Q

What is the ESR

A

erythrocyte sedimentation rate or sed rate
to detect inflammatory conditions, infections, malignancies

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25
Q

What is the principle of an ESR

A

distance in mm the RBCs fall per 1 hour

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26
Q

What are the disadvantages to the ESR test

A

low sensitivity and specificity

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27
Q

What factors affect ESR

A

RBC factors: rouleaux, RBC mass proportional to EST
plasma factors: too many proteins cause rouleaux,
mechanical factors: tilted tube, insufficient centrifuge

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28
Q

Indicate whether the following examples would have an increased or a decreased ESR
Pregnant patient 
Anemic patient (NOT sickle cell) 
Sickle cell anemia  doesn’t increase the sed rate, could be normal, or decreased
Polycythemia 
Newborn 
Multiple myeloma (increased proteins in blood)

A

Pregnant patient - increased
Anemic patient (NOT sickle cell) - increased
Sickle cell anemia  doesn’t increase the sed rate, could be normal, or decreased
Polycythemia - decreased
Newborn - decreased
Multiple myeloma (increased proteins in blood)- increased

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29
Q

Who most often does POC testing

A

nurses

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30
Q

What are 3 POC tests

A

hematocrit on hematostat
Hgb conentration- HemoCue
cell and plt counts on heme analyzer

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31
Q

What is electronic impedance

A

detection and measurement of changes in the electrical resistance produced by cells as they cross a small space on instryment

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32
Q

What separates cells in electronic impedance

A

volume thresholds

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33
Q

What does the height of voltage pulse =

A

volume of cell

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34
Q

How are results given in impedance tests

A

volume distribution histogram

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35
Q

What type of differential in impedance tests

A

3 part diff
lymphs
mononuclear cells and granulocytes

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36
Q

What is radiofrequency

A

low voltage DC impedance that is used with high voltage electromagnetic radiofrequencies

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37
Q

Cell volume is proportional to ______

A

change in DC

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38
Q

Cell interior density is proportional to

A

change in RF signal

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39
Q

What results do radiofrequency instruments give

A

scatterplots

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40
Q

What kind of differential do radiofrequency instruments give

A

5 part diff
neutrophils, lymphs, monocytes, eosinophils and basos

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41
Q

Forward scatter light at 0 degrees relates with

A

cell volume

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42
Q

sideway scatter light relates with

A

internal complexity of the cell

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43
Q

What instrument utilizes MAPSS to perform its WBC dif?

A

cell-DYN

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44
Q

New parameters and indices may be useful for

A

diagnosis and treatment of anemias

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45
Q

Immature retic fraction or IRF shows

A

an early indication of erythropoiesis

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46
Q

IFR and absolute retic counts

A

help distinguish types of anemias

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47
Q

Match: cold agglutinins, icterus and lipemia, hemolysis, plt clumps, old specimen

-low RBs and high HCT,
-high MCV and MCHC and decreased RBC
- high MCV, high MPV, low plt
-high HGB and high MCH
-low plts high WBCs,

A

hemolysis- redraw
cold agglutinins- warm specimen
old specimen- redraw
icterus and lipemia- plasma replacement
plt clumps- sodium citrate tube

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48
Q

How much time do we have to make a blood film slide

A

4 hrs

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49
Q

What angle should the pusher slide be at

A

30-45 degree

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50
Q

How should the angle change if there is a high HCT? and a low one?

A

high- lower angle
low- higher angle

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51
Q

How much of the slide should the film take up
what shape should it have

A

2/3
feathered edge

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52
Q

What stain is used in peripheral blood smears

A

wright Geimsa stain

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53
Q

What do polychrome stains contain

A

eosin and methylene blue

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54
Q

What does eosin do, is it acidic or basic

A

acidic, stains cell components like Hgb, eosinophillic granules

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55
Q

What does methylene blue do? is it acidic or basic

A

stains acidic cellular components like RNA
it is basic

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56
Q

List the steps to staining a blood smear slide

A

flood with wright stain for 1-3 min
add buffer to slide, must look metallic green, 3 min
rinse with distilled water and air dry in vertical position

57
Q

What should the colors of RBCs, WBCs and eosinophils have on a proper stained blood film

A

RBCs orange to pink
WBCs purple nuclei , pink cytoplasm and granules visible
eosinophils bright orange

58
Q

What causes RBCs to appear gray, WBCs to be dark and eosinophils to look gray on a blood film

A

stain or buffer was too alkaline
did not rinse stain well enough
stained for too long
used heparin specimen for CBC

59
Q

What causes RBCs to be too pale or red
and WBCs to be barely visible

A

stain or buffer is too acidic, not enough buffer, rinsed too much

60
Q

what is the ideal area for reading a peripheral smear

A

a spot where RBCs are uniformly distributed, no touching or overlapping, normal biconcave appearance
200-250 RBCs per field

61
Q

What units are used to report
WBC
RBC
HGB
HCT
PLT

A

SI listed first
common listed second

HGB and HCT- Common units, not in L- in uL or dL or %

RBC, WBC, PLT- SI unit, in L

62
Q

What is absolute vs relative

A

absolute- real numbers
relative is a percentage

63
Q

philia means ____ and cytosis means ____

A

philia- increase
cytosis- also increase

64
Q

penia mean

A

decrease

65
Q

RDW, HGB, HCT, MCV, MCH, MCHC are all parameters of

A

RBCs

66
Q

What results could be given if RDW is out of range

A

variations in RBC volume
anisocytosis- if high
11.5-14.5%

67
Q

If a sample has an MCV of 70?
and 110?

A

70- microcytic
110-macrocytic

68
Q

If a sample has an MCHC of 36
or of 20

A

20-hypochromic
36- spherocytes

69
Q

MPV is a CBC parameter for

A

platelets
mean platelet volume

70
Q

an increased plt count is called ____
a decreased plt count is called ____

A

thrombocytosis
thrombocytopenia

71
Q

If a sample has increased plts, WBCs and HGB this is called
if all are decreased it is called

A

pancytosis
pancytopenia

72
Q

If WBC count is low it is called __
if it is low ___

A

leukopenia
leukocytosis

73
Q

The best parameter that indicates a pt is anemic is

A

the HGB

74
Q

If a sample has abnormally large platelets

A

the platelet histogram will be abnormal with peaks

75
Q

What is the reason flow cytometry testing is performed

A

detection of fluorescent tags directly bound to specific molecules or through monoclonal antibodies

75
Q

What is the reason flow cytometry testing is performed

A

detection of fluorescent tags directly bound to specific molecules

76
Q

What is flow cytometry most often used for

A

leukemias, lymphomas, monitoring HIV

77
Q

What test tubes are used in flow cytometry?

A

heparin and EDTA

78
Q

How much time do you have to process a sample into flow cytometry after the time of collection

A

24-48 hrs

79
Q

How should specimen for flow cytometry be stored

A

less than 24 hrs at RT for BM and peripheral blood
less than 24hrs at 4C for tissues and fluids

80
Q

What allows a sample to be stained with flurochrome?

A

monoclonal antibodies

81
Q

What lineage do these CD markers represent
CD34
CD117
TdT

A

Immature cells

82
Q

What lineage do these CD markers represent
CD33, CD13, CD15, CD14

A

granulocytes/ monocytes

Three blind Mice

83
Q

What lineage do these CD markers represent
CD71, glycoprotein A

A

erythroid

Rrolling stones, 70s rock

84
Q

What lineage do these CD markers represent
CD41, CD42, CD61

A

Megakaryocytic
Four-fat albert

85
Q

What lineage do these CD markers represent
CD19, CD20, CD22, kappa light chain, gamma light chain

A

B lymphs
Barbie in her 20s

86
Q

What lineage do these CD markers represent
CD2,3,4,5,7,8

A

T- lymphs
Think of cells at work immature T.

87
Q

consists of fluidics, a light source (laser), a detection system, and a computer
Cells must pass one by one through the illumination and detection system

A

flow cytometer

88
Q

central core of individually aligned cells is surrounded by a sheath fluid, get them in single file line
Central alignment is essential for consistent illumination of cells as they pass before a laser light source

A

hydrodynamic focusing

89
Q

What is forward scatter vs side scatter

A

forward- volume/ size
side- complexity

90
Q

Explain how Flow cytometry works

A

cells are injected into a fluid and placed in a single file stream, they emit a color when exposed to a laser, forward and side scatter are detected

91
Q

electronic boundary used by an operator to delineate cell clusters

A

gate

92
Q

a process of selecting a population of interest as defined by one or more flow cytometry parameters

A

gating

93
Q

acquisition of data for a specific cell pop as define by flow parameters

A

live gating

94
Q

What is the CD45 marker

A

pan hematopoitic antigen

95
Q

how much CD45 marker density for
lymphs
granulocytes
erythroid precursors

A

lymph-high density
granulocytic- intermediate
erythroid precursors- none

96
Q

What cells have these markers
CD34, CD117, CD38 and HLA-DR

A

pluripotent stem cells

97
Q

How are hematologic neoplasms diagnosed and classified with flow cytometry

A

based on hematopoietic maturation pathways

98
Q

What cell is characterized as having low density expression of CD45

A

blasts

99
Q

How to ID mature B and T cells
and mature lymphoid neoplasms?

A

surface immunoglobulin receptors, polyclonal pop

neoplasm- monoclonal B or T receptors

100
Q

What is the CD4:CD8 ratio in health individuals

A

> 1

101
Q

What is the CD4:CD8 ratio in HIV positive pts

A

reversed due to decrease in CD4 lymphs

102
Q

PNH CD55 DAF, CD59 MIRL markers are decreased

A

RBCs

103
Q

PNH CD24 and FLAER markers are decreased

A

granulocyte antigens

104
Q

PNH CD14 and FLAER are decreased

A

monocyte antigen

105
Q

What are the purine and pyrimidine nucleotides

A

pure-AG
pyrimid- CUT

106
Q

Why is the isolation of RNA more difficult than the isolation of DNA

A

RNA is harder because of presence of ribonucleases RNases on mammalian epithelial surfaces

107
Q

What is the principal technique in the clinical molecular laboratory

A

PCR

108
Q

Why is it so important to prevent contamination when using molecular testing methods

A

RNAses- can amplify anything that contaminates the test, on any mammalian cell

109
Q

enzyme that disrupts hydrogen bonds between DNA strands to allow them to separate for DNA replication

A

helicase

110
Q

enzyme method for amplifying a specific target sequence to allow its detection even in small amounts

A

PCR

111
Q

What is the purpose of PCR

A

amplification of DNA

112
Q

End point vs real time PCR

A

end-amplicon detected at the end
real time- amplicon detected during each PCR cycle

113
Q

Put into order
-first cycle of PCR complete
- repeat 25-40 times
-DNA poly binds first nucleotide to primer
-DNA denatured 95C
-cool to 40-60C, primers anneal to complimentary strands

A

-DNA denatured 95C
-cool to 40-60C, primers anneal to complimentary strands
-DNA poly binds first nucleotide to primer
-first cycle of PCR complete
- repeat 25-40 times

114
Q

What is the purpose of RT-PCR

A

amplification of RNA

115
Q

What is the starting material of RT-PCR

A

mRNA

116
Q

The rate at which DNA fragments migrate through gels is proportional to the mass only not their relative charge

A

in gel electrophoresis

117
Q

fragments sieved through an agarose or polyacrylamide gel matrix by passing through the gel as it is bathed in a conducting salt solution

A

the process of gel electrophoresis

118
Q

smaller or larger
Agarose pores are ___
polyacrylamide gel pores are _____

A

larger
smaller

119
Q

anode vs cathode

A

anode-positive charge
cathode-negative

120
Q

If there is a mutation present, restriction endonucleases will not cleave the same sites, and will produce restriction fragments of different lengths than normal

A

RFLP restriction fragment length polymorphism

121
Q

measures the change in nucleic acid amplification as replication progresses using fluorescent marker dyes

A

real time PCR

122
Q

what types of body fluids can be analyzed in heme

A

CSF
serous or body cavity
synovial
bronchoalveolar lavage BAL

123
Q

What is the volume of each side of the hematocytometer

A

9mm^3

124
Q

yellow color to supernatant of CSF

A

xanthochromia

125
Q

What tube is the cell count performed on

A

tube 3

126
Q

the cells that line CNS, can be seen in CSF diffs
appear in clumps, look like malignant .

A

ependymal and choroid plexus cells

127
Q

Cells in CSF if vertebral body is accidentally punctured

A

cartilage cells

128
Q

macrophages that have ingested RBCs and have resulted in the breakdown of RBCs causing hemosiderin to be released

A

siderophages

129
Q

large rough dark blue or black granules in cytoplasm

A

hemosiderin

130
Q

golden yellow figures

A

bilirubin crystals

131
Q

What does the presence of siderophages indicate

A

a pathologic hemorrhage

132
Q

lining cells of body cavities, fried egg appearance, 1-3 nucleoli

A

mesothelial cells

133
Q

intact neutrophil that has engulfed homogenous mass of degenerated nuclear material, which displaces the normal nucleus

A

lupus erythematosus cell

134
Q

What cells are normal in synovial fluid

A

lymphs, monos/ histiocytes, synovial cells

135
Q

can be seen in the upper respiratory tract, abnormal deeper inside the lungs,

A

ciliated epithelial cells

136
Q

What cells shouldnt be seen in a BAL fluid

A

mesothelial cells

137
Q

What forward and side scatter amounts do lymphs, monos and granulotcytes have

A

lymphs- small- low forward scatter, simple- low side scatter
monos-large- high forward scatter, complex medium side scatter
granulocytes- medium- medium forward scatter, high complexity- medium side scater

Hematology (4 decks)

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