To study of total RBC count in human blood.
Apparatus: Haemocytometer,sterilized prcking needle,r.b.c pipette, compound microscope and hayems diluting fluid ( Nacl-1%,na2so4-2.5%,and Hgcl2)
#Haemocytometer
=Haemocytometer is a kind of slide containing counting chambers.
=The Burker, Neubaur,thoma or any other Haemocytometer may be used.
=The Neubaur Haemocytometer consists of a double -cell slide with two sunk platforms in open cell type.
=Each platform has a rulling so that rapid duplicate counts can be made.
=When a special cover glass is kept in position,a depth of 1/10 mm is maintained over the rulligs.
=Diluted blood Is run on by capillary force after the cover glass has been fixed.
=Gunting chambers are nine large square and each medium sides of1mm each.
=Central large square ABCD is subdivided in 25 medium squares and each medium in turn contain 16 tiny squares having sides 0.05 mm long.
=Hayens diluting fluid may be kept as stock solution and a small amount of it may be taken in watch glass,so that entire solution may not be mixed with the blood.
@procedure
= Clean and dry mixing pipette,(dilution 1:200)
=Sterilise your middle finger and the puncturing needle(preferably over a flame ) with a a small pad of cotton wool dipped in 90% alcohol.
=Pick the middle finger with the puncturing needle so that blood flows freelt .don't squeeze.
=Wipe away clanly the first few drops ,then suck the blood with RBC pipette upto the0.5 mark slowly and carefully. If the excess May be removed by wiping the end of the pipette suitably drying on blotting paper.
=Wipe the excess blood from the tip of the pipette and immediately mix it with hayens diluting fluid suck in RBC pipette the diluting blood upto 101mark hayens fluid prevents haemolysis,roulequx formation, coagulation and bacterial growth. Disconnect the rubber tube ,grip the end of the pipette between forefinger and thbnd Shake throughly for a minute.
=Re - attach the rubber tube to the pipette reject the unused diluting fluid in the stem of the pipette. Avoid drying and quickly run the diluted blood under the cover glass on to each of the central platforms . If unable to do so, wash the side and the cover glass and repeat it until you become expert. The acquirement of skill in performing procedure no 3and 6 very essential and otherwise the counting will be valueless.The correct size of the blood drop and speed is obligatory, in orderto avoid uneve distribution of the cell.very large drop will result in low count because the cells tend to be sucked down into the through away from the counting grid on the contrary,a very small drop,so that a second drop is to be added,result in high count due to the uneven distribution of cells at the junction of the two drops.once an optimum drop has been placed, the cell should be allowed to settle for are minute and then counting should be done.
=Bring counting scale into focus under The objective under a microscope. Count cells in 16sq. In five different parts of the field. The counting should be done exact replica of the sqa droen on your copy any corpuscle lying on lines should be moved either upward s on the right side.
#Calculation
These may be done in the following manner.
Number of r.b.c. per cubic mm=number of cells counted×Dilution×100
--------------------------------------------------------------------number of small counted
#Result
Total number of R.B.C. counted 1mm=5.46×10'6mm.
To study of total RBC count in human blood.
Apparatus: Haemocytometer,sterilized prcking needle,r.b.c pipette, compound microscope and hayems diluting fluid ( Nacl-1%,na2so4-2.5%,and Hgcl2)
#Haemocytometer
=Haemocytometer is a kind of slide containing counting chambers.
=The Burker, Neubaur,thoma or any other Haemocytometer may be used.
=The Neubaur Haemocytometer consists of a double -cell slide with two sunk platforms in open cell type.
=Each platform has a rulling so that rapid duplicate counts can be made.
=When a special cover glass is kept in position,a depth of 1/10 mm is maintained over the rulligs.
=Diluted blood Is run on by capillary force after the cover glass has been fixed.
=Gunting chambers are nine large square and each medium sides of1mm each.
=Central large square ABCD is subdivided in 25 medium squares and each medium in turn contain 16 tiny squares having sides 0.05 mm long.
=Hayens diluting fluid may be kept as stock solution and a small amount of it may be taken in watch glass,so that entire solution may not be mixed with the blood.
@procedure
= Clean and dry mixing pipette,(dilution 1:200)
=Sterilise your middle finger and the puncturing needle(preferably over a flame ) with a a small pad of cotton wool dipped in 90% alcohol.
=Pick the middle finger with the puncturing needle so that blood flows freelt .don't squeeze.
=Wipe away clanly the first few drops ,then suck the blood with RBC pipette upto the0.5 mark slowly and carefully. If the excess May be removed by wiping the end of the pipette suitably drying on blotting paper.
=Wipe the excess blood from the tip of the pipette and immediately mix it with hayens diluting fluid suck in RBC pipette the diluting blood upto 101mark hayens fluid prevents haemolysis,roulequx formation, coagulation and bacterial growth. Disconnect the rubber tube ,grip the end of the pipette between forefinger and thbnd Shake throughly for a minute.
=Re - attach the rubber tube to the pipette reject the unused diluting fluid in the stem of the pipette. Avoid drying and quickly run the diluted blood under the cover glass on to each of the central platforms . If unable to do so, wash the side and the cover glass and repeat it until you become expert. The acquirement of skill in performing procedure no 3and 6 very essential and otherwise the counting will be valueless.The correct size of the blood drop and speed is obligatory, in orderto avoid uneve distribution of the cell.very large drop will result in low count because the cells tend to be sucked down into the through away from the counting grid on the contrary,a very small drop,so that a second drop is to be added,result in high count due to the uneven distribution of cells at the junction of the two drops.once an optimum drop has been placed, the cell should be allowed to settle for are minute and then counting should be done.
=Bring counting scale into focus under The objective under a microscope. Count cells in 16sq. In five different parts of the field. The counting should be done exact replica of the sqa droen on your copy any corpuscle lying on lines should be moved either upward s on the right side.
#Calculation
These may be done in the following manner.
Number of r.b.c. per cubic mm=number of cells counted×Dilution×100
--------------------------------------------------------------------number of small counted
#Result
Total number of R.B.C. counted 1mm=5.46×10'6mm.
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