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MLSCI 330 > Automated Testing > Flashcards

Automated Testing Flashcards

1
Q

pulse height analyzer

A

oscilloscope
- sorts and categorizes the pulses and plots them on a frequency distribution graph or volume distribution graph

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2
Q

RBC histogram

A

36-360 fL
curve should begin at baseline
smaller population normally to the right of the main population = cells that have been corrected for coincidence

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3
Q

PLT histogram

A

2-20 fL
curve of best fit overlaid on raw data
curves start and stop at baseline

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4
Q

WBC histogram

A

> 35 fL
should start or be really close to baseline
three distinct populations usually (lymphs, monos, grans)

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5
Q

the principle by which hematology instrumentation provides white blood cells differentials and nRBC differentiation

A

flow cytometry

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6
Q

sample types for flow cytometry

A
  • routine = EDTA whole blood
  • special heme = EFTA whole blood, LiHeparin whole blood
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7
Q

clinical applications of flow in special heme

A
  • T cell enumeration: monitoring progression of diseases such as HIV, and other immunodefs
  • Lymphoma/Leukemia investigations: determining origin and ID of immature/abnormal cells
  • immunophenotyping for PNH and hereditary spherocytosis
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8
Q

what does the diluent do in beckman/sysmex?

A
  • dilutes & stabilizes whole blood sample
  • conduct aperture current
  • focus sample stream
  • rinse instrument between runs
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9
Q

hemoglobinometer in beckman

A
  • lysing contains cyanide pigment that binds and stabilizes the free hemoglobin
  • once WBC count is complete, solution passed onto spec/hemoglobinometer
  • stable cyanide-hemoglobin pigment measured at 525 nm
  • %transmittance is compared to reagent blank
  • Hb concentration calculated using Beer’s Law and reported g/L
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10
Q

Beckman VCS technology

A
  • volume analysis = low frequency
  • conductivity analysis = high frequency current differentiate between insulating cell properties (nuclear, granular, chemical composition of cell interior)
  • light scatter analysis = cell complexity and granularity
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11
Q

5 part diff reported by Beckman

A

neuts monos lymphs basos and eos

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12
Q

T or F. Beckman instruments are abl to report nRBCs based off VCS technology

A

T

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13
Q

how does the beckman instrument differentiate nRBCs from WBCs?

A

VCS technology

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14
Q

nRBCs are often mistaken as these

A

small lymphs (by Beckman)

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15
Q

supravital stain for retics

A

new methylene blue

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16
Q

what does new methylene blue do to retics?

A

precipitates nucleic acids inside immature RBC (increasing granularity)

17
Q

T or F. More immature RBC = more residual RNA and greater volume

A

T! larger and higher light scatter
(appear more to the right of a dot plot)

18
Q

derived parameters in Beckman

A
  • MCV: average vol of all RBCs; fL
  • RDW: coefficient of variation of RBC Vol distribution (spread of gaussian distribution form histogram); %
19
Q

calculated parameters in Beckman

A
  • Hct: relative vol of packed RBCs; Ht is roughly 3x the Hb value if RBCs are normochromic; L/L
  • MCHC: ave weight of Hb in a sample
  • MCH: weight of Hb in average RBC
20
Q

which instrumentation uses hydrodynamic focusing?

A

Sysmex
- cells sheathed by diluent and passed through the aperture one at a time to prevent coincidence and recirculation of red cells

21
Q

how are cells distinguished based on size in Sysmex?

A

floating thresholds for the cell population (determined by the instrument)
- this allows for discrimination of populations on a sample-by-sample basis
- also helps separate PLTs from small red cells

22
Q

when is a fluorescent PLT done?

A

when there are interferences in the impedance PLT count
- fluorescent markers label PLT RNA (after perforation)
- PLTs separated using FSC and fluorescence
- this method can differentiate immature PLT and mature PLT based on RNA

23
Q

flow chamber in sysmex

A
  • enumerate WBCs, nRBCs and the 6-part diff
  • sample and diluent taken into WNR channel where RBCs (including nRBCs) are lysed
  • fluorescent polymethine dye added to stain nucleus and organelles of WBC w/ high intensity and stains the extruded nRBC nucleus with low intensity
24
Q

T or F. Unlike Beckman, WBCs are counted as they are differentiated in the flow cell in the Sysmex

A

T! it’s also corrected for nRBC

25
Q

what does SFL detect

A

side fluorescent light determined if nucleic acids are present inside the cell

26
Q

6th part of diff in Sysmex

A

immature grans

27
Q

RET channel in sysmex

A
  • lysis reagent added to perforate RBC, WBC, and PLT membrane
  • fluorescent makes label nucleic acids in the cell
  • FSC and SFL differentiate RBC from retics
28
Q

when are manual retic counts with NMB performed?

A
  • interference flags on automated counts
  • relative >30%
  • absolute >90 x10^9/L
  • at least two reticulin granules must be present in the cell to be denoted a retic
29
Q

hemoglobinometer in sysmex

A
  • Hb oxidized and sodium lauryl sulfate added to form complex
  • measured at 555 nm
  • %T compared to reagent blank
30
Q

derived parameters in sysmex

A
  • Hct
  • RDW

MLSCI 330 (9 decks)

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