APPARATUS: ay CHAPTER 9. TOTAL LEUCOCYTIC COUNT (TLC) Counting chamber, WBC diluting pIPert® (White bead), WBC diluting thulds: of diluting Hlulds are used In TEC: Two type: 1) Gincial acetic acid= 2m! 19¢nqueous sol. of gentian violet = tent m Distilled water= 100m! 2)NNO HCI dic in naturo and cause ysis of Rag, 8. Those diluting Mulds are acl PROCEDURE: FILLING THE vsnelet . the WI mark in ‘BC diluting pipette, which is smaller and hy Draw the blood up to 0.8 white bead In the bulb. Wipe the end & reduce the column of blood up to 0.5 ark if it has gone gone above, 2 3) Draw the diluting fluid slowly up te mark 17 #bove the bulb, this mak i eae ata . this makes a dilution of 1:20. 1) Discard 2-3 drops from the pipette before filling the counting ch 2) Allow at least one minute for erythrocyte lysis and for the leuk see 3) Under low power count the number of cells In each of the 4 1, vor cnines wa 4) The rules for counting the WBC touching the lines is same os tort . 5) Anuneven distribution of cells is indicated when ther vain 2 ne i Ua tablet vounee iS a Variation of more than 15 cells CALCULATION: i No of WBCs counted in four squares X Dilution factor X Volume correct Wit)+ WI2} + WI3) 4 WI) X 20 X 2.5= leucocytes x10°a/ul ientmet SOURCES-OF ERROR: 1) Error in dilution. 2) Eroy4n counting, 5) Failure to wipe excess hlood from tipf pipette. 3) Error in calculation goon. shaking of oi pipette 6) Overcharging of chamber. 7) Improper adjustment of light source, which ma Y make the leuko cytes difficult to ob: serve ©” Temperature 2) 4) Trauma 5) Neoplasia . keris DECREASED VALUE: rosie Shalit, 4) Leucopenia Human (Male) (Female) 5---10 5----B,