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HEMATOLOGY 1 > Flow cytometry and Cytometry > Flashcards

Flow cytometry and Cytometry Flashcards

1
Q

Most common clinical application of flow cytometry

A

Diagnosis of leukemias and lymphomas

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2
Q

Flow cytometry’s original purpose

A

To evaluate physical properties of cells based on their ability to deflect light

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3
Q

Discovery that improved flow cytometry

A

Development of monoclonal antibodies

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4
Q

Particles studied in flow cytometry

A

Cells, chromosomes, microorganisms, proteins

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5
Q

Properties measured by flow cytometry

A

Physical, antigenic, and functional properties of particles suspended in a fluid

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6
Q

Components of a flow cytometer

A

Fluidics, light source (laser), multiple detectors, computer

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7
Q

Main advantage of flow cytometry

A

Ability to quickly and simultaneously analyze multiple parameters in a large number of cells

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8
Q

Specimens commonly analyzed by flow cytometry

A

Bone marrow, peripheral blood, lymphoid tissues, body cavity fluids, solid tissues

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9
Q

Processing time for peripheral blood and bone marrow specimens

A

Must be processed within 24 to 48 hours from time of collection

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10
Q

Preferred anticoagulant for specimens in flow cytometry

A

Heparin

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11
Q

Flow cytometry specimen transportation

A

Transported at room temperature

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12
Q

Meaning of LASER

A

Light Amplified by Stimulated Emission of Radiation (Photon is the basic unit of radiation)

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13
Q

Flow cytometry antigen detection

A

Can detect 17 antigens on an individual cell simultaneously using monoclonal antibodies conjugated to fluorochromes

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14
Q

Flow cytometry analysis method

A

Cells must pass separately through the illumination and detection system

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15
Q

Method for individual cell analysis

A

Hydrodynamic focusing (cells aligned in a core surrounded by sheath fluid)

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16
Q

Fluorescence detection in flow cytometry

A

Particles emit fluorescent signals detected by detectors, converted to digital output

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17
Q

Additional signals recorded in flow cytometry

A

Forward scatter (FS), Side scatter (SS), and fluorescence

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18
Q

Forward scatter (FS) detection

A

Measures particle volume or size, detected by a photodetector aligned with the laser beam

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19
Q

Side scatter (SS) detection

A

Measures surface complexity and internal structures (granules and vacuoles), detected by a photodetector positioned to the side

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20
Q

Display and registration of flow cytometry signals

A

FS, SS, and fluorescence displayed simultaneously on the instrument screen and registered by the computer system

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21
Q

Markers for Immature lineage

A

CD 34, CD 117, Terminal deoxynucleotidyl transferase

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22
Q

Markers for Granulocytic/Monocytic lineage

A

CD 33, CD 13, CD 15, CD 14

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23
Q

Markers for Erythroid lineage

A

CD 71, Glycophorin A

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24
Q

Markers for Megakaryocytic lineage

A

CD 41, CD 42, CD 61

25
Q

Markers for B lymphocytes

A

CD 19, CD 20, CD 22, κ Light chain, λ Light chain

26
Q

Markers for T lymphocytes

A

CD 2, CD 3, CD 4, CD 5, CD 7, CD 8

27
Q

Study of chemical constituents of cells

A

“Cytochemistry definition

28
Q

Enzymatic technique specimen requirement

A

Fresh smears

29
Q

Nonenzymatic technique specimen stability

A

Stable for months at room temperature (e.g., PAS/SBB)

30
Q

Enzymes seen in primary granules of neutrophils

A

eosinophils

31
Q

Use of MYELOPEROXIDASE (MPO)

A

Used to differentiate blasts of AML from ALL

32
Q

Positive MPO stain interpretation

A

Rules out ALL

33
Q

Positive MPO results

A

Neutrophilic granulocytes (except normal blasts), Auer rods, leukemic blasts in FAB M1, M2, M3, eosinophils

34
Q

Weakly positive or negative MPO results

A

Monocytes

35
Q

Negative MPO results

A

Myeloblasts, basophils, lymphocytic cell series, erythrocytic cell series

36
Q

Stain reactions parallel those of MPO’s in most cases

A

“SUDAN BLACK B (SBB)”

37
Q

SUDAN BLACK B (SBB) stains

A

Sterols, neutral fats, phospholipids (found in primary and secondary granules of neutrophils and lysosomal granules of monocytes)

38
Q

Positive SBB results

A

Promyelocyte, myelocyte, metamyelocytes, bands, segmented neutrophils (STRONGLY POSITIVE), leukemic blasts, Auer rods, eosinophils

39
Q

Weakly positive or negative SBB results

A

Myeloblasts, monocytic cells

40
Q

Negative SBB results

A

Lymphocytes and its precursors, megakaryocytes and platelets, erythrocytes

41
Q

Stain to differentiate acute granulocytic leukemias from monocytic leukemias

A

“ESTERASES”

42
Q

Esters substrates

A

α-naphthyl acetate, α-naphthyl butyrate (nonspecific), Naphthol AS-D Chloroacetate (specific)

43
Q

α-naphthyl acetate esterase (NSE) positive results

A

Monocytes (strong positive reaction), positive for certain cell types

44
Q

α-naphthyl acetate esterase (NSE) weakly positive or negative results

A

Granulocytes, lymphoid cells

45
Q

α-naphthyl acetate esterase (NSE) negative results

A

Monocytes (with NaF inhibition)

46
Q

α-naphthyl butyrate esterase (NSE) positive results

A

Monocytes, positive for certain cell types

47
Q

α-naphthyl butyrate esterase (NSE) weakly positive or negative results

A

Granulocytes, lymphoid cells

48
Q

α-naphthyl butyrate esterase (NSE) negative results

A

Monocytes (with NaF inhibition)

49
Q

Naphthol AS-D Chloroacetate esterase positive results

A

Promyelocyte, myelocyte, metamyelocyte, bands, segmented neutrophils, leukemic myeloblasts, Auer rods

50
Q

Naphthol AS-D Chloroacetate esterase weakly positive or negative results

A

Monocytic cells

51
Q

Naphthol AS-D Chloroacetate esterase negative results

A

Myeloblasts (variable), monoblasts, promonocytes, monocytes

52
Q

Stain for glycogen

A

“PERIODIC ACID-SCHIFF (PAS)”

53
Q

PERIODIC ACID-SCHIFF (PAS) is useful in identifying

A

“FAB M6 leukemia”

54
Q

Strongly positive Leukemia using PERIODIC ACID-SCHIFF (PAS) REACTION

A

“RBCs in erythroleukemia (FAB M6)”

55
Q

Normal erythroid precursors reaction in PAS

A

“negative”

56
Q

Enzyme marker for primitive lymphoid cells

A

“TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE (TdT)”

57
Q

Strong TdT activity – observed in approximately 90% of patients with

A

“ALL”

58
Q

Helpful in the recognition of the ‘lymphoblastic transformation’ of CML

A

“TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE (TdT)”

HEMATOLOGY 1 (27 decks)

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