Hematology Instrumentation SOLO 1

0 degrees angle scatter

• axial light loss

• measures cell size

7 degrees angle scatter

• intermediate angle scatter

• measures cell complexity

90 degrees angle scatter

• polarized side scatter

• measures lobularity

90D degrees angle scatter

• depolarized side scatter

• measured granularity

Beckman Coulter IRIS

• consists of two smaller instruments joined together

  • IRIS iChem velocity

  • IRIS iQ-200 sprint

• can perform fully automated urinalysis for a more streamlined lab and minimal intervention from the tech

IRIS iChem velocity

performs biochemical, specific gravity, color, and clarity analysis

IRIS iQ-200

• measured urine sediment similar to a flow cytometer

• measures size, shape, contrast, and texture

• uses Auto Particle Recognition to classify particles

• takes up to 500 pictures/second

RBC/platelet histogram

• counted by optical light scatter

• measured for MCV/MPV using optical technology

• RDW/PDW calculated

WBC differential scatterplot

plots ALL(cell size) against IAS(cell complexity)

WBC differential scatterplot region 1

• normal blood

  • few events in the region

• abnormal blood

  • lyse resistant RBCs

  • sickle cells

  • fetal cells

  • nRBCs

WBC differential scatterplot region 2

• normal blood

  • lymphs and monos

• abnormal blood

  • atypical lymphs

  • blasts

WBC differential scatterplot region 3

• normal blood

  • few to no cells

• abnormal blood

  • atypical monocytes

  • blasts

  • plasma cells

WBC differential scatterplot region 4

• normal blood

  • few to no cells

• abnormal blood

  • immature granulocytes

  • neutrophil aggregates

  • platelet sattelitism

Low-Hi Fl scatterplot

• relative amount of intact membranes

• plots ALL (cell size) against fluorescence (FL1)

mono poly 1 scatterplot

• specializes in platelet aggregates

• plots PSS (lobularity) versus ALL (cell size)

reticulocyte scatterplot

• relative ratio of retics to mature RBCs

• indirectly measures erythropoietic and megakarypoietic activity

What are falsely increased in aged blood?

MCV and HCT

What is falsely increased in lipemia?

MCHC and HGB parameters

What are CBC characteristics of cold agglutinin?

invalid data and elevated MCHC; decreased RBC and HCT

What characteristics are affected by thalassemia?

low MCV and HGB

What are the characteristics of sickle cell?

• low RBC, HGB, HCT, MCV

• increased RDW and RBC frag flag

What are the characteristics of AML?

• high white count

• invalid data, left shift, var lym, and blast flags

impedance counting

• a cell suspension is created using an electrically conductive system

  • requires a conductive liquid

• a cathode and anode are placed in opposite sides of the imped chamber

• a decrease in current indiciates a cellular event is counted

• the amount of current decrease is inversely proportional to the approximate size

Alinity H uses what methods to count blood

• photometry/absorbance (HGB)

• optical light scatter (WBC, RBC, PLT, NRBC counting; WBC class and diff)

• fluorescence (detection of WBC, nRBC, RETC, and REC PLT)

absorbance spectrophotometry

• a cells suspension of RBCs are mixed with a lysing agent

• released hemoglobin from inside RBCs reacts with the reagent to form a colored complex that absorbs 540nm light

• the amount of light absorbed is proportional to the amount of hemoglobin present

Light functions in what three ways that is useful for analysis?

• diffraction

• refraction

• change in speed

• reflection

AlinityH achieves accurate optical light scatter analysis with:

• advanced MAPSS

  • advanced multi-angle polarized scatter separation

• detects scattered light at 4 angles using a blue diode laser

  • counts RBCS, WBCs, PLTs,nRBCs, and RETCs

  • detects fluorescence from WBCs, nRBCS, and RETCs

  • ccharacterizes WBCs for diff

IAS1

• measures hemoglobin concentration

• size of PLTs

IAS2

• measures size of RBCs

• MCV from average of IAS2

IAS3

• distinguishes PLT from RBC

• distinguishes PLT from noise

fluorescence detection

• fluorochromes are added to different dilutions and attach to specific cellular components

• 530nm light signal labeled as FL1

Reticulocyte fluorescence detection

• dye binds to RNS

• helps detemine compensatory ability of bone marrow

nRBC fluorescence detection

dye binds to DNA

WBC vaibility/differential fluorescence detection

• dye binds to DNA

• as WBCs age, cell membrane degrades and fluorochrome enters cells and attaches to DNA

h

hydrodynamic focusing

• ensures orderly passage of cells through a sensing zone

• moves with laminar flow which prevents mixing

• prevents clogging

alinity requires what anticoagulant?

K2-EDTA

Alinity minimum volume

340 microliters whole blood

Alinity h system consists of what major components?

• analyzer

• reagents

• data station/computer

• printer

Alinity hq reagents

• diluent/sheath reagent (diluting/rinsing)

• hemoglobin reagent

• WBC reagent

• reticulocyte reagent

• water from WPS

• autoclean reagent

cyanide free hemoglobin reagent

• primes HGB reagent tubing

• rinses HGB system

• lysing agent

• reduces interference from leukocytes by detroying

• converts HGB to a single chromagen (540nm)

alinity hs reagents/supplies

• smear fix

• wright-giemsa stain

• phosphate buffer

• glass slides

• smear tape cartridge

• slide label cartridge

What qualities does the iQ-200 use to analyze microscopic urine sediment?

• texture

• shape

• size

• contrast

What analyses does the iChemi velocity perform?

• biochemical

• clarity

• specific gravity

• color

list the tests present on a urine reagent strip

• glucose

• blood

• nitrite

• protein

• ketone

• pH

• specific gravity

• urobilinogen

• bilirubin

• leukocyte esterase

Three major systems of flow cytometers

• fluidics

• optics

• electronics

FACS stands for

fluorescence activated cell sorting

What part of the microscope functions to focus and center the light beam with a desired numerical aperture and field size through the specimen?

condenser

parts of the microscope are divided into what categories?

magnification and illumination