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Hematology 2 Lab > LABORATORY ACTIVITY 8 > Flashcards

LABORATORY ACTIVITY 8 Flashcards

1
Q

Last stage of immature RBC

A
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2
Q

Stage between the

A

orthochromatophilic normoblast and a mature erythrocyte

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3
Q

maturation time in the bone marrow:

A

2-3 days

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4
Q

maturation time in the circulation:

A

1 day

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5
Q

First normal immature cell seen in the circulation at a very low value

A
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6
Q

Presence of other immature cell indicates

A

dysplasia, cytopenia, etc.

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7
Q

: with remnants/web/hairlike fragments; w/o nucleus

A

retic

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8
Q

youngest RBC with no nucleus

A

Retic

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9
Q

: very clear pink cytoplasm; w/o remnant; w/o nucleus

A

rbc

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10
Q

: w/ nucleus

A

orthochromatophilic normoblast

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11
Q

Cytoplasmic inclusions coprecipitate with the few remaining [?] to for visually stained [?] with a supravital stain

A

mitochondria and ferritin masses (also RNA remnants)

dark/blue clusters

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12
Q

: leaves RNA once precipitated

A

Mitochondria (+nucleus & ribosome)

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13
Q

: completely disappears with the nucleus

A

DNA

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14
Q

: slow to precipitate (left behind) = RNA remnants

A

RNA

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15
Q

: iron stores; precipitates in the cytoplasm and binds with the remnants (RNA + iron); used in Hb production

A

ferritin

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16
Q

cytoplasmic inc:

A

RNA precipitates (mostly) and Iron precipitates

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17
Q
  • a dye that stains living cells and its inclusions; dark blue clusters
A

Supravital stain

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18
Q

: smear → air dry → stain (methylene blue, eosin, dist water, alcohol)
Preservation: (dies)

A

RBC

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19
Q

: EDTA-blood is mixed w/ stain → incubation → stain
No preservation: (living)

A

Retic

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20
Q

: remnants are not visible

A

Wright stain/Polychrome-stain

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21
Q

: remnants are visible

A

New methylene blue, Brilliant cresyl blue

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22
Q

Specimen:

A

 WB mixed w/ any anticoagulant

 Capillary (w/o anticoagulant)

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23
Q

– preferred; preserves size, shape; least contamination/fragmentation/precipitation

A

EDTA

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24
Q

Safer (allows multiple smears)

A

EDTA

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25
Q

o Collected on microtainer w/ graduations

A

Capillary (w/o anticoagulant)

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26
Q

o 1:1 (stain:blood)

A

Capillary (w/o anticoagulant)

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27
Q

o performed on babies

A

Capillary (w/o anticoagulant)

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28
Q

o requires smearing technique (limited to 3 smears)

A

Capillary (w/o anticoagulant)

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29
Q

SUPRAVITAL STAINS

A

 New methylene blue and Brilliant cresyl blue

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30
Q

Must be filtered daily or before use

A

 New methylene blue and Brilliant cresyl blue

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31
Q

may precipitate in amber bottle (confusion w/ remnant/contamination

A

 New methylene blue and Brilliant cresyl blue

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32
Q

PROCEDURE:

A

DRY PREPARATION

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33
Q
  1. Mix equal amounts of [?] in a small test tube.
A

filtered stain and EDTA-anticoagulated blood or fresh capillary blood

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34
Q

blood : stain

A

1 ml : 1 ml

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35
Q
  1. Incubate mixture at [?]
A

room temperature for 10-15 minutes.

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36
Q

Rodaks: 3-10 mins; Minimum: 10 mins; Maximized for [?] in the lab to stain properly

A

15 mins

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37
Q

Inversion:

A

8x

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38
Q

Remix the tube after [?] (to prevent settling or rbc and stain at the bottom of the tube = no retics will be seen)

A

15 mins

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39
Q
  1. After incubation, mix thoroughly and prepare a [?]
A

wedge smear

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40
Q

 Prepare at least [?] smears (1 for each stain)
 Air dry

A

2

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41
Q
  1. Examine smear using [?]. Select an area where erythrocytes are close but not overlapping and reticulocytes appear to be well-stained.
A

100x objective

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42
Q

 Criteria:

A

PASS (size, length, presence of bubble, dropping marks, consistency of the tail from thick to thin)

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43
Q

: scanning (to avoid rouleaux; tail)

A

10x

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44
Q

: confirm whether rbc or retic

A

40x

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45
Q

: counting

A

100x

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46
Q
  1. Count the number of reticulocytes in [?] red cells. Reticulocytes should also be counted as erythrocytes.
A

1000

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47
Q

 10 fields with [?] rbc each in the lab

A

100

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48
Q

 At least [?] is retic in the circ

A

1.5%

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49
Q
  1. Calculate as follows:
A
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50
Q

: no Hct and maturation day (count only)

A

Uncorrected

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51
Q

● measure of erythropoietic ability of the bone marrow in response to anemia

A

RETICULOCYTE COUNT

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52
Q

With retic:

A

normal rbc production

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53
Q

Indications:

BM transfusion

A

Anemia

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54
Q

Adult:

A

0.5 - 2.5% of total RBC

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55
Q

Newborn:

A

4.0 - 6.0% of total RBC

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56
Q

Circulation:

A

0.-5 to 1.5%

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57
Q

Decreased with aging

A

Adult: 0.5 - 2.5% of total RBC

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58
Q

Higher bm activity

A

Newborn: 4.0 - 6.0% of total RBC

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59
Q

: anemia = the retic prod is decreased

A

<0.5

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60
Q

: erythrocytosis = the retic prod is increased

A

> 1.5

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61
Q

37 retics are found when 1000 erythrocytes are examined (37 retics, 963 erythrocyte). What is the retic count?

A

3.7%

Interpretation: Increased production of reticulocyte.

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62
Q

● Traditional way of counting that reduces the cells to count

A

MILLER OCULAR DISC

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63
Q

A disc inserted into the eyepiece

A

MILLER OCULAR DISC

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64
Q

Permits a less labor-intensive surveying of RBCs

A

MILLER OCULAR DISC

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65
Q

Count at least 112 RBCs in successive fields (CAP)

A

MILLER OCULAR DISC

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66
Q

MILLER OCULAR DISC Determine the retic ct using this formula:

A
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67
Q

MILLER OCULAR DISC
2 squares:

A

o A (large square) - for counting retics
o B (small square) - for counting RBC

68
Q

INCREASED

A

Blood loss

Crisis associated with HA

Subsequent treatment for PA

Folate and iron deficiency

Hypoxia

69
Q

DECREASED

A

Aplastic anemia

Aplastic crisis of HA

Chemotherapeutic radiation and induced hypoproliferation

Pernicious anemia

Dec erythropoiesis

70
Q

: retics are released early from the bm = ↑ retic to compensate for the loss

A

 Blood loss

71
Q

: rbc is destroyed from the circulation due to and antigen causing hemolysis = ↑ retic

A

 Crisis associated with HA

72
Q

: treatment of intrinsic factor = = ↑ retic

A

Subsequent treatment for PA

73
Q

: lack of oxygen

A

Hypoxia

74
Q

hiking in very high altitude: dec oxygen intake due to dec production of rbc

A

Hypoxia

75
Q

↓ rbc ct, ↓oxygen, ↓ rbc production = ↓ EPO

A

Hypoxia

76
Q

: no rbc

A

 Aplastic anemia

77
Q

: hemolysis of both rbc and retic

A

 Aplastic crisis of HA

78
Q

: bm cannot produce rbc and rectic; blast cell is also affected

A

 Chemotherapeutic radiation and induced hypoproliferation

79
Q

: intrinsic factor deficiency in hydrochloric acid produced by the stomach

A

 Pernicious anemia

80
Q

absorbs Vit B 12; helps in absorption of Vit B 12 and Folic acid for the maturation of rbc

A

 Intrinsic factor

81
Q

: bm is not produced efficiently = dec retics

A

Dec erythropoiesis

82
Q

Retic differentiates

A

anemias

83
Q

Causes of anemia:

A
  1. Problem in the bm
  2. Increased destruction
84
Q

defective production = anemia

A
  1. Problem in the bm
85
Q

cause: hormone, genetic, chemotherapy

A
  1. Problem in the bm
86
Q

abnormal rbc = decreased retic

A
  1. Problem in the bm
87
Q

normal bm but destructed in circ

A
  1. Increased destruction
88
Q

cause: antigen, viral, para, bacte infection

A
  1. Increased destruction
89
Q

decreased survival of rbc

A
  1. Increased destruction
90
Q

normal rbc = increased retic

A
  1. Increased destruction
91
Q

Hallmark of anemia:

A

increased retic

92
Q

SORCES OF ERROR

A

Interobserver variation in the definition of a retic
Accuracy of counting of the observer (±20%)
Refractile bodies due to poor drying
Precipitates
Others:
-Severe anemia
-Failure to re-mix after incubation, prior to smear preparation
-Bias in the use of Miller Disc grid

93
Q

o atmospheric moisture may confuse remnant w/ refractile bodies

A

Refractile bodies due to poor drying

94
Q

o may resembke dark blue clusters in retic filter stain

A

Precipitates

95
Q

o confuse remnant w/ contaminats

A

Precipitates

96
Q

o solution: filter the stain before use

A

Precipitates

97
Q

o Proportion of dye-to-blood must be adjusted accordingly

A

Severe anemia

98
Q

1 ml of blood : 0.5 ml stain

A

Severe anemia

99
Q

 use the similar inverted L rule applied in hemocytometers

A

Bias in the use of Miller Disc grids

100
Q

INCLUSIONS CONFUSED WITH RETICS

A

Howell-Jolly bodies

Heinz bodies

Pappenheimer bodies

101
Q

: 1-2 deep purple-colored, dense structures

A

Howell-Jolly bodies

102
Q

irregular circular inclusion (not web-like)

A

Howell-Jolly bodies

103
Q

: light-blue green (not blue)

A

Heinz bodies

104
Q

usually on the periphery of the rbc

A

Heinz bodiesv

105
Q

: small iron clusters

A

Pappenheimer bodies

106
Q

most common confusion

A

Pappenheimer bodies

107
Q

o Solution: prepare 2 smears (retic and wright-stained)

A

Pappenheimer bodies

108
Q

o use prussian cytochemical stain blue (preferred) over wright-giemsa

A

Pappenheimer bodies

109
Q

: more effective to differentiate Pappenheimer from retic remnant

A

o use prussian cytochemical stain blue (preferred) over wright-giemsa

110
Q

FALSELY DECREASED

A

Understaining (cause: improper incubation)
High glucose levels (can be mask the appearance of RNA remnants)

111
Q

(cause: improper incubation)

A

Understaining

112
Q

(can be mask the appearance of RNA remnants)

A

High glucose levels

113
Q

Absolute Reticulocyte Count (ARC) FORMULA

A
114
Q

Absolute Reticulocyte Count (ARC) RV

A

20-115x109 /L

115
Q

Corrected Reticulocyte Count (CRC) FORMULA

A
116
Q

Corrected Reticulocyte Count (CRC) RV

A

0.5-1.5% (adults) 4.0-0.6% (newborn)

117
Q

If Hct = 35%, CRC is typically at

A

25%

118
Q

If Hct = <25%, CRC is typically at

A

3-5%

119
Q

Reticulocyte Production Index (RPI) FORMULA

A
120
Q

In anemic patients, RPI should be

A

> 3

121
Q

If RPI [?] , adequate bm response is seen

A

> 3

122
Q

If RPI [?], an inadequate bm response is seen

A

<2

123
Q

40-45

A

1

124
Q

35-39

A

1.5

125
Q

25-34

A

2

126
Q

15-24

A

2.5

127
Q

Expression of retic count out of 1000 rbc

A

Uncorrected/Relative Reticulocyte Count (URC/RRC)

128
Q

Bias for patients with very low Hct (packed rbc)

A

Uncorrected/Relative Reticulocyte Count (URC/RRC)

129
Q

Actual number of reticulocytes in a 1 L of blood

A

Absolute Reticulocyte Count (ARC)

130
Q

More preferred method of reporting reticulocyte counts

A

Absolute Reticulocyte Count (ARC)

131
Q

More accurate and reliable than URC

A

Absolute Reticulocyte Count (ARC)

132
Q

Actual counting of number of retic expressed through the rbc

A

Absolute Reticulocyte Count (ARC)

133
Q

Opposite of URC

A

Corrected Reticulocyte Count (CRC)

134
Q

Calculated to account for the degree of anemia by using a standard normal Hct of 45% (0.46 L/L)

A

Corrected Reticulocyte Count (CRC)

135
Q

URC cannot be reported alone (may be normal out of 1000 rbc, but prbc may be low in actual)

A

Corrected Reticulocyte Count (CRC)

136
Q

Correction factor: 45% Hct (to correct the degree of anemia using hematocrit)

A

Corrected Reticulocyte Count (CRC)

137
Q

are often present during compensation of anemia

A

Shift reticulocytes

138
Q

(w/ more RNA remnants)

A

Shift reticulocytes

139
Q

Reticulocytes that have transitioned or shifted from the bone marrow to the circulation earlier than usual

A

Shift reticulocytes

140
Q

If [?] is seen in a stained PBS, corrections should be made to account for shift retics.

A

polychromasia

141
Q

indication of shift retics

A

polychromasia

142
Q

gray-pink, salmon pink, gray, very red

A

Retics w/ polychromasia

143
Q

Px hematocrit %, Correction factor (?)

A

maturation time or days

144
Q

during cases of anemia, shift retics increase tocompensate for [?] (↓ rbc = no oxygen)

A

hypoxia

145
Q

maturation: 2-3 days (bm) ; 1 day (circulation)
 Anemia: release from the bm earlier than 2-3 days
 To correct the presence of shift reticulocytes

A

Reticulocyte Production Index (RPI)

146
Q

shift retics often mature loger than normal usually takes 3 days

A

Reticulocyte Production Index (RPI)

147
Q

CF must be with reference to the Hct value of the patient

A

Reticulocyte Production Index (RPI)

148
Q

Example: Retic ct: 7.8%, Hct (%) = 30%; Ans: 2.6%

A

Reticulocyte Production Index (RPI)

149
Q

Purpose: To determine an adequate bm response in cases of anemia

A

Reticulocyte Production Index (RPI)

150
Q

Solution: transplant the blood or transplant the bm

A

Reticulocyte Production Index (RPI)

151
Q

OTHER METHODS

A
152
Q

 had been widely used nowadays

A
  1. Flow cytometry
153
Q

 automated: provides absolute and relative counts (unlike in CRC and RPI)

A
  1. Flow cytometry
154
Q

 can be assessed using automated methods

A
  1. Immature Reticulocytes Fraction (IRF)
155
Q

 Counts retics with more RNA (immature/shift)

A
  1. Immature Reticulocytes Fraction (IRF)
156
Q

 Provides CRC and RPI

A
  1. Immature Reticulocytes Fraction (IRF)
157
Q

 assessment of erythropoietic activity after chemotherapy or stem cell transplantation (to asses bm function)

A
  1. Immature Reticulocytes Fraction (IRF)
158
Q

 is analogous to the RBC MCH

A
  1. Reticulated Hemoglobin Content (CHr)
159
Q

o measurement of Hb in retic

A
  1. Reticulated Hemoglobin Content (CHr)
160
Q

 can be used to detect early cases of iron deficiency

A
  1. Reticulated Hemoglobin Content (CHr)
161
Q

o MCH: content Hb of the cell (hypochromic)

A
  1. Reticulated Hemoglobin Content (CHr)
162
Q

Reticulocyte count is very important in determining whether the anemia is caused by

A

(1) defect in erythropoiesis, (2) hemolysis/ shortened survival

163
Q

there is an adequate release of the bone marrow

A

defect in erythropoiesis

164
Q

there is a normal production of RBC but RBCs die in the circulation

A

hemolysis

165
Q

In this case, napipilitan si bone marrow na labas ng labas ng reticulocyte. Thus, mas nag compensate yung bone marrow kay (?) in defect in erythropoiesis.

A

hemolysis/ shortened survival

166
Q

a hallmark of anemia.

A

increased reticulocyte

167
Q

● Same goes with blood loss when injured, there is also an (?) because the bone marrow needs to compensate for the RBC that was lost.

A

increased reticulocyte

Hematology 2 Lab (7 decks)

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