CBC Expt - phyana lab Flashcards
Universal precaution
Treat all body fluids infectious
Tests in CBC
Hemoglobin measurement Hematocrit measurement RBC count WBC count Differential Count
MEASURES Volume of packed red cell after centrifugation
HEMATOCRIT measurement
METHOD IN hematocrit measurement
Adam’s micromethod
materials in hematocrit measurement
capillary tube (red, blue) sealing clay/paraffin wax microhematocrit centrifuge machine microhematocrit reader
Procedure in hematocrit measurement
Fill capillary tube with blood (3/4) Seal. Label Centrifugate. Use balancer Spin 10,000 rpm 4-5min
in hematocrit measurement, spin at what rpm and how many min
10,000 rpm 4-5min
Hematocrit NT
male- 47 +/- 7 vol% / 0.47 +/- 7 female- 42 +/- 5 vol%
Methods for hemoglobin measurement
acid-hematin method Cyanmethemoglobin method
Method for hemoglobin measurement that gives less error
Cyanmethemoglobin method
MATERIALS FOR hemoglobin measurement
0.01 N HCl Sahli’s tube (w/comparator block) Sahli’s pipette dH20 Stirring rod
PROCEDURE FOR hemoglobin measurement
Deliver 0.01N HCl into Sahli’s tube – 2mark Aspirate 0.02 mL blood Expel blood into Sahli’s tube Rinse pipette with dH20-add into the tube Add dH20 drop by drop – comparator block Read lower miniscus
NR hemoglobin measurement
NR: male : 14-16.5 g/100mL or 14-16.5g% 140-165 g/L female: 12-15g/100mL
Used in counting RBC, WBC, Platelet, absolute eosinophil count, sperm count With thick cover slip Improved Neubauer Counting Chamber
hemacytometer
specific hemacytometer in the lab
Improved Neubauer Counting Chamber
of WBC in 1 cu mm of blood Dil fluid must lyze RBC
WBC count
materials for WBC count
WBC pipette Dil fluid hemacytometer
procedure for wbc count
Aspirate blood 0.5 mark. Wipe sides Aspirate RBC dil fluid up to 101 mark. Mix dil’n 1:200 Discard 5-6 gtt Charge counting chamber Stand 5min Count RBC in HPO
materials for differential count
Blood 2 slides Differential stain Pasteur pipette Cedar wood oil
High refractive index, thereby increasing the numerical aperture of the objective lens Have specific optical and viscosity characteristics
cedar wood oil
describe cover glass used in differential count
thicker, less flexible, surface tension of the fluid will not deform them, height of the fluid is standardized
procedure for differential count
Prepare smear Dry smear on flat surface Stain smear Count 100 cells in OIO
technique used in differential count
strip differential technique
how many WBC must be counted in a smear
100
devices used for DIFFERENTIAL COUNT
hand tally counter differential counter
how many quadrants are there for WBC
4
What are counted in WBC or RBC
dots
how many secondary squares are in improved neubauer
9
how many tertiary squares are in 1 WBC square
16
what is the total number of tertiary squares of a WBC
16 x 4 = 64
1 RBC square = ___ tertiary squares
5
1 RBC tertiary square =___ small squares
16
total number of small RBCsquares
16 x 5 quadrants = 80 small squares
what are the staining solutions in differential count (list in order)
1st: ethanol
2nd: eosin
3rd: methylene blue
4th: buffer or distilled water
staining solution for fixing the slide
ethanol
staining solution for staining
eosin
methylene blue
staining solution for rinisng solution or stains
buffer or distilled water
