Clinical Chemistry: Practical 2

what is FSC

forward scatter size

what is SSC

side scatter size

what does forward scatter tell us

size

what does side scatter tell us

complexity

what are the specimens analyzed in flow cytometry

blood

bone marrow

biopsies

what are the fluorophores used for flow cytometry

FITC and PE

what does FITC stand for

fluorescein isothiocyanate

what does PE stand for

phycoerythrin

one dimension plot for flow cytometry

histogram

two dimension plot for flow cytometry

scatter plot

Type 1 scatterplot for flow cytometry

tells us what cell markers are and are not there. four gated quadrants

what does the bottom left of type 1 flow cytometry scatter plot tell us

neither X or Y

what does the bottom right of type 1 flow cytometry scatter plot tell us

only X

what does the top left of type 1 flow cytometry scatter plot tell us

only Y

what does the top right of type 1 flow cytometry scatter plot tell us

both X and Y

what does the type 2 flow cytometer scatter plot show us

displays complexity and one cell marker. shows what kinds of cells are in the sample

what does purple mean on type 2 flow cytometry scatterplot

neutrophils

eosinophils

what does blue mean on type 2 flow cytometry scatterplot

monocyte

what does green mean on type 2 flow cytometry scatterplot

lymphocyte

what does red mean on type 2 flow cytometry scatterplot

blasts

cell markers for granulocytes

CD13

CD14 (+/-)

CD 33

cell markers for monocytes

CD13

CD14

CD33

cell marker for RBC

CD71

cell marker for megakaryocyte

CD61

cell marker for B cell lymphocytes

CD19

CD20

CD10

cell marker for T cell lymphocytes

CD2

CD3

CD5

CD7

+CD4

+CD8

CD4 is a cell marker for

helper cells

CD8 is a cell marker for

cytotoxic cells

cell marker for NK cells

CD16

cell marker for progenitor cells

CD34

laminar flow describes

orderly flow of cells

hydrodynamic focusing describes

cells going through one at a time

sweep flow describes

making sure cells go through only once

what does diacetyl monoxime method measure

BUN

what is the reaction for the diacetyl monoxime method

Serum urea + diacetyl monoxide + strong acid → Diazin (yellow chromogen) measured at 540 nm

BUN color reagents

diacetyl monoxime thiosemicarbazide

BUN acid reagents

ferric chloride hexahydrate

sulfuric acid

phosphoric acid

what is the purpose of the heating block in diacetyl monoxime

to accelerate the reaction

what is the purpose of the ice bath in diacetyl monoxime

to stop the reaction and stabilize the color complex

what is the calculation for diacetyl monoxime

(unknown ABS/standard ABD) X standard concentration = unknown concentration

what specimens are used in diacetyl monoxime

serum

plasma

urine

reagents for diacetyl monoxime

Serum urea + diacetyl monoxide + strong acid

result of the diacetyl monoxime reaction

Diazin (yellow)

what nm is diacetyl monoxime measured at

540 nm

what is the purpose of measuring BUN

to assess kidney function and detect possible renal or metabolic disorders.

BUN reference range

6-20 mg/dL

Creatinine reference range

0.6-1.2 mg/dL

BUN/creatinine ratio reference range

10-20

pre renal causes of increased BUN

diet, protein breakdown, protein synthesis

renal causes of increased BUN

glomerulonephritis

nephritis

nephrotic syndrome

renal necrosis

renal failure

post renal causes of BUN

tumor

gallstone

kidney stone

inflammation

when BUN/CREAT ratio is normal

renal cause

when BUN/CREAT ratio is abnormal

pre renal

post renal

specimen used for the Jaffe reaction

serum

reagents for the Jaffe reaction

creatinine

picric acid

NaOH (alkaline solution)

byproduct of Jaffe reaction

Jaffe chromogen (orange red)

what wavelength is the jaffe chromogen detected at

510 nm

what is the purpose of measuring creatinine

evaluate kidney function and monitor for renal impairment

Jaffe reaction

serum + picric acid + NaOH (alkaline) --> jaffe chromogen

reference range for creatinine

0.6-1.2 mg/dL

what could cause a high creatinine level

decreased kidney function

what could cause low creatinine levels

malnutrition

what is a control

mix of analytes making a sample similar to what is in the body which has a known range of concentrations.

what is a standard

single analyte of a known concentration

How to remember the Jaffe reaction?

Jaffe picks orange red in alkaline conditions

how to remember the diacetyl monoxime reaction?

diacetyl turns into yellow acidic diazin