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HEMATOLOGY MODULE 1 & 2 > MODULE 2 AUTOMATION > Flashcards

MODULE 2 AUTOMATION Flashcards

1
Q

Two basic cell counting principles employed in most hematology analyzers

A

Electrical Impedance
Optical Scatter or Detection

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2
Q

What is Hydraulics

A

aspirating unit, dilutors, dispenser, mixing chambers, aperture baths, flow cells and hemoglobinometer

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3
Q

What is PNEUMATICS

A

Vacuums and pressure for opening valves and moving the sample through the system

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4
Q

What is ELECTRICAL SYSTEMS

A

Electronic analyzers and computing circuity for processing data

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5
Q

What is Electrical Impedance

A

Also know as low-voltage direct current resistance
most commonly methodology used
example are Coulter Counter and Sysmex Counter

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6
Q

Principle of Electrical Impedance

A

Cell passes through the aperture, cells do not conduct current but they change electrical resistance which is counted as voltage pulses.

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7
Q

Size Threshold Ranges in an Electrical Impedance Method

A

`RBCs 36-360 fL
WBCs 45-450 fL
Lymphocytes 45-90 fL
Monocytes 90-160 fL
Granulocytes 160-450 fL
Platelets 2-20 fL

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8
Q

Principle of OPTICAL SCATTER

A

Differentiate and enumerate cell types based on the scattering properties of the cells.
Patterns of scatter are measured in various angles
Uses laser and nonlaser light

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9
Q

What are the Patters of Scatters are measured by various angles

A
  1. Forward angle light source
  2. Side angle light scatter
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10
Q

What is Forward angle light scatter

A

measure cell size

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11
Q

What is Side angle light scatter

A

Measures cell granularity and lobularity

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12
Q

Side angle light scatter consist of

A

a. RBC no nucleus
b. WBC with nucleus

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13
Q

Example of LIGHT used in Optical Scatter

A

a. Tungsten
b. Helium-neon laser

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14
Q

what is Tungsten

A

halogen lamp

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15
Q

Helium-neon Lamp

A

Laser monochrome light

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16
Q

What is a Laser Light

A

most common light source used in flow cytometers because of its properties INTENSITY, STABILITY and MONOCHROMATISM

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17
Q

Principle of FLOW CYTOMETRY

A

Cells suspension is run under high pressure in single, narrow stream of laser, causing excitation of flourescent compounds resulting emission of light energy. Energy is detected by a photomultiplier tube and converted into computerized data, which provide information regarding the number, size and cellular components.

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18
Q

Major components of Flow Cytomers

A

Fluidics
Optics
Electronics

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19
Q

What is FLUIDICS

A

flow chamber for single cell seperation, sheath fluid and hydrodynamic focusing

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20
Q

what is OPTICS

A

excitation of light source includes lasers or lamps.
Light is seperated by dichroic mirror and filters

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21
Q

example of lasers

A

argon
krypton
Helium-neon
helium cadium
diode

(KAHHD)

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22
Q

examples of Lamps

A

Mercury
Xenon-mercury

(MX)

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23
Q

what is Electronics

A

Photomultuplier tube detecs light energy, then converts this to voltage pulses, computers translate pulses into data files.

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24
Q

Benefits of Optical Technology

A

Laser Light can be focused on individual cells
More than 2 measurements can be made and more information can be gathered
Cells are passed in a single file through the flow cells
More realistic results

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25
Q

Principle of histogram

A

utilizes impedance technology and representation of cell number versus one measured property, usually cell size.
Produced by plotting the number on the y axis and cell size on the x axis

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26
Q

How many distribution peaks is the Normal WBC Histogram

A

3 Distribution Peaks

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27
Q

First Peak WBC Histogram

A

45-90 fL: small mononuclear cells (lymphocytes

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28
Q

Second Peak WBC Histogram

A

90-160 fL: minor population of large mononuclear cells (monocytes, reactive lymphocytes, immature WBCs)

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29
Q

Third Peak WBC Histogram

A

160-450 fL: normal mature types of granulocytes

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30
Q

WBC categories on Coulter Counter

A

Small Cells (lymphocytes)
Medium Cells (Reactive lymphocytes)
Large Cells (Granulocytes)

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31
Q

Abnormal WBC Histogram

A
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32
Q

Enumerate Abnormal WBC Histogram

A

R1: <35 fL indicate nRBCs, giant and clumped platelets
R2: 90 fL indicate Reactive lymhocytes or blast cells
R3: 160 fL indicate increase in bands, immature neutrophils, eosinophils or basophils
R4: >450 fL indicate high granulocyte and multiple region overlap (RM)

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33
Q

Enumerate Abnormal WBC Histogram

A

R1: <35 fL indicate nRBCs, giant and clumped platelets
R2: 90 fL indicate Reactive lymhocytes or blast cells
R3: 160 fL indicate increase in bands, immature neutrophils, eosinophils or basophils
R4: >450 fL indicate high granulocyte and multiple region overlap (RM)

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34
Q

Abnormal WBC Histogram

A
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35
Q

Other flags in WBC Histogram

A

H: parameter value is higher than set normal limit
L: parameter value is lower than set normal limit

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36
Q

Other flags in WBC Histogram

A

H: parameter value is higher than set normal limit
L: parameter value is lower than set normal limit

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36
Q

Abnormal WBC Histogram

A
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37
Q

RBC Histogram

A

6-360 fL or >36 fL
Show single peak at between 70 and 110 fL

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38
Q

Abnormal RBC Histogram Interpretation

A

24-36 fL Reject (RBC Histogram can measure cells as small as 24 fL)
Shift to the Right: Macrocytic
Shift to the Left: Microcytic
Bimodal, 2 Peaks: Cold agglutinin, hemolytic anemia with schistocytes present, anemia with different cell size (when patient present blood transfusion, dimorphic RBC population)
Increased Curved Width: Anisocytosis (correlate with increase RDW)

39
Q

WBC Histogram

A

24-36 fL Reject (RBC Histogram can measure cells as small as 24 fL)
Shift to the Right: Macrocytic
Shift to the Left: Microcytic
Bimodal, 2 Peaks: Cold agglutinin, hemolytic anemia with schistocytes present, anemia with different cell size (when patient present blood transfusion

40
Q

WBC Histogram

A

24-36 fL Reject (RBC Histogram can measure cells as small as 24 fL)
Shift to the Right: Macrocytic
Shift to the Left: Microcytic
Bimodal, 2 Peaks: Cold agglutinin, hemolytic anemia with schistocytes present, anemia with different cell size (when patient present blood transfusion

41
Q

WBC Histogram

A

24-36 fL Reject (RBC Histogram can measure cells as small as 24 fL)
Shift to the Right: Macrocytic
Shift to the Left: Microcytic
Bimodal, 2 Peaks: Cold agglutinin, hemolytic anemia with schistocytes present, anemia with different cell size (when patient present blood transfusion

42
Q

Platelet Histogram

A

2-20 fL

43
Q

Platelet Histogram Interpretation

A

Lower Region Interference: <2 fL Electrical Impedence
Upper Rehion Interference: >20 fL Microcytic RBCs, Schistocytes, Giant or Clumped Platelets

44
Q

Abnormal RBC Histogram Interpretation

A
45
Q

Scattergram

A

also called CYTOGRAM OR SCATTERPLOT
2-dimensional representation of 2 or more cell properties or characteristics plotted against each other

46
Q

Scattergram

A

also called CYTOGRAM OR SCATTERPLOT
2-dimensional representation of 2 or more cell properties or characteristics plotted against each other

47
Q

Scatter plots for WBC

A

Displayed on a monitor and colored coded for different subpopulations

48
Q

Methodologies of Scattergram

A

Radiofrequency
Fluorescence
Cytochemistry

49
Q

Problems Encountered in Automated Methods

A

Instrumental Errors
Nature of the Specimen

50
Q

Example of Instrumental Error

A

Positive Errors
negative Errors
Improper Settings of aperture current/threshold

51
Q

What is a Positive Errors

A

Aperture

52
Q

Examples Positive Errors

A

Aperture plug- most common problem in cell counting
Bubbles in the sample- cause by vigorous mixing
Extraneous electrical pulses

53
Q

Example of Negative Errors

A

Excessive lysing of RBCs

54
Q

Example of Improper setting of aperture current/threshold

A

cause either + or - errors

55
Q

Example of Nature of specimen error

A

Giant platelets
Fragment of WBC cytoplasm may be counted as RBCs or Platelets
Agglutination of RBCs, WBCs and Platelets
Platelet Sattelitism

56
Q

Effect of Giant Platelets in Nature of Specimen Errors

A
57
Q

Effect of Giant Platelets in Nature of Specimen Errors

A

counted as RBCs or WBCs

57
Q

Effect of Giant Platelets in Nature of Specimen Errors

A

counted as RBCs or WBCs

58
Q

Effect of Fragment of WBC Cytoplasm in Nature of Specimen Errors

A
59
Q

Effect of Fragment of WBC Cytoplasm in Nature of Specimen Errors

A

counted as RBCs or Platelets

60
Q

Effect of Agglutination of RBCs, WBCs and Platelets in Nature of Specimen Errors

A

False-negative results for each count

61
Q

Effect of Fragment of WBC Cytoplasm in Nature of Specimen Errors

A

counted as RBCs or Platelets

61
Q

Effect of Agglutination of RBCs, WBCs and Platelets in Nature of Specimen Errors

A

False-negative results for each count

62
Q

Effect of Platelets Satellitism in Nature of Specimen Errors

A

Falsely low platelets count

63
Q

Cold agglutinins Parameters Affected

A

INCREASE: MCV, MCHC
DECREASE: RBC
Grainy Appearance

64
Q

Corrective action for Cold Agglutinins

A

warm specimen 37 degree Celsius and RERUN

65
Q

Corrective action for Cold Agglutinins

A

warm specimen 37 degree Celsius and RERUN

66
Q

Lipemia Icterus Parameters Affected

A
67
Q

Lipemia Icterus Parameters Affected

A

INCREASE: Hb, MCH

68
Q

Corrective action for Cold Agglutinins

A

warm specimen 37 degree Celsius and RERUN

69
Q

Lipemia Icterus Parameters Affected

A

INCREASE: Hb, MCH

69
Q

Lipemia Icterus Parameters Affected

A
70
Q

Hemolysis Parameters Affected

A

DECREASE: RBC, Hct

70
Q

Corrective action Lipemia Icterus

A

Plasma replacement

71
Q

Corrective action Hemolysis

A

Request new specimen

72
Q

Lysis-resistant RBCs Parameters Affected

A

INCREASE: WBC, Hb

73
Q

Corrective action Lysis-resistant RBCs

A

Perform manual dilution, allow incubation time for lysis

74
Q

Microcytes or Schistocytes Parameters Affected

A

INCREASE: PLT
DECREASE: RBC

75
Q

Corrective action for Microcytes or Schistocytes

A

review blood film

76
Q

nRCS, Megakaryocytes fragments or Micromegakaryoblasts Parameters Affected

A

INCREASE: WBC (older specimen)

77
Q

Corrective action for nRCS, Megakaryocytes fragments or Micromegakaryoblasts

A

Newer instruments
Count nRBCs and correct WBC count
Count micromegakaryoblasts per 100 WBCs and correct

78
Q

Platelets Clumps Parameters Affected

A

INCREASE: WBC
DECREASE: PLT

79
Q

Corrective action for Platelets Clumps

A

Redraw specimen in Sodium Citrate, multiply result by 1.1

80
Q

WBC, 100.000/mL Parameters Affected

A

INCREASE: Hb, RBC, Hct

81
Q

Corrective action for WBC, 100.000/mL

A

Manual Hct
Manual Hb

82
Q

Leukemia, especially with chemotherapy Parameters Affected

A

INCREASE: PLT
DECREASE: WBC

83
Q

Corrective action for Leukemia, especially with chemotherapy

A

Review film
Perform phase platelet count or CD61 count

84
Q

Old Specimen Parameters Affected

A

INCREASE: MCV, MPV
DECREASE: PLT

85
Q

Corrective action for Old Specimen

A

establish stability and specimen rejection criteria

86
Q

Lyse resistance abnormal RBCs

A

Sickle Cells
Target Cells
Hypochromic Cells

(STH)

87
Q

Criteria to repeat the analysis if

A

Rule of 3 failures on a normochromic sample
Any results outside linearity limits established by the manufacturer- dilute into linearity range
unexplained delta check failures

87
Q

Effect of Agglutination of RBCs, WBCs and Platelets in Nature of Specimen Errors

A

False-negative results for each count

88
Q

Criteria to repeat the analysis if

A

Rule of 3 failures on a normochromic sample
Any results outside linearity limits established by the manufacturer- dilute into linearity range
unexplained delta check failures

88
Q

Effect of Agglutination of RBCs, WBCs and Platelets in Nature of Specimen Errors

A

False-negative results for each count

HEMATOLOGY MODULE 1 & 2 (3 decks)

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