3 Blood cells – under microscope NEUEOLYMBASOMONONRBCPLTRBC
4 Hematology parameters 1 White blood cell – WBC(pcs/l, pcs/µl)Number of leukocytes.WBC = WBCcal x (pcs/l or pcs/µl)Red Blood Cell – RBCNumber of erythrocytes.RBC = RBCcal x (pcs/l or pcs/µl)Haemoglobin concentration - HGB(g/dl, g/l, mmol/l)Measured photometrically at 540 nm; in each cycle blank measurement is performed on diluentHGB = HGBcal x (HGBmeasured – HGBblank)Mean Corpuscular Volume - MCV(fl=10-15 l)Average volume of individual erythrocytes derived from the RBC histogram.Haematocrit – HCT(percentage, absolute)Calculated from the RBC and MCV values.HCTpercentage = RBC x MCV x 100,HCTabsolute = RBC x MCVMean Corpuscular Haemoglobin – MCH(pg, fmol)Average hemoglobin content of erythrocytes, calculated from RBC and HGB values.MCH = HGB / RBC
5 Hematology parameters 2 Mean Corpuscular Haemoglobin Concentration – MCHC(g/dl, g/l, mmol/l)Calculated from the HGB and HCT values.MCHC = HGB / HCTabsoluteUnit of measurement is displayed according to the one chosen for HGB result (g/dl, g/l or mmol/l)Red cell Distribut. Width - RDW-SDandPlatelet Distribut. Width - PDW-SD(fl)Red cell Distribut. Width - RDW-CVPlatelet Distribut. Width - PDW-CV (absolute)The distribution width of the erythrocyte or platelet population derived from the histogram at 20% of peakxDW-SD = RDWcal x (P2 – P1) (fl),xDW-CV = RDWcal x 0.56 x (P2 – P1) / (P2 + P1)by the factor of 0.56 CV is corrected to the 60% cut
6 Hematology parameters 3 Platelet – PLT(pcs/l, pcs/µl)Number of thrombocytes (platelets)PLT = PLTcal x (pcs/l or pcs/µl)Mean Platelet Volume – MPV(fl)Average volume of individual platelets derived from the PLT histogram.Thrombocrit – PCT(percentage, absolute)Calculated from the PLT and MPV values.PCTpercentage = PLT x MPV x 100,PCTabsolute = PLT x MPVWhite blood cell differential:LYM#, LY% : lymphocytesMID#, MID%: monocytes and some eosinophil granulocytesGRA#, GR%: neutrophil, eosinophil and basophil granulocytesAbsolute values counted in the channels determined by the three WBC discriminators:Percentages calculated from the absolute WBC value.1.: RBC-LYM discriminator2.: LYM-MID discriminator3.: MID-GRA discriminator1.2.3.
10 Coincidence effect >1 cells in aperture look one big cell Less counts, distorted histogramSolution: diluting samples, mathematical coincidence correctionLinearity range (80um):WBC: 100 x 10^3/ulRBC: x 10^6/ulPLT: 1000 x 10^3/ul
14 Hemoglobin measuring principle Specific wavelength light source – green 560 nmLight to frequency converterAmbient light compensation with U/D counting, and light choppingLED SAMPLE DETECTOR LED switching:Frequency output
15 Hemoglobin calculation – HGB blank HGB needs blank measurement on clean diluent to compensate temperature driftHGB = C * ln (CNTblank / CNTsample)CNTblank = HGB count on diluent =CNTsample = HGB count on sample =