Rajagopalan Prabhu
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Hemacytometer Cell Counting Logic- by Prabhu (M.Tech Ph.D)
red square = 1.0000 mm2, 100.00 nl
green square = 0.0625 mm2, 6.250 nl
yellow square = 0.040 mm2, 4.00 nl
blue square = 0.0025 mm2, 0.25 nl
at a depth( height) of 0.1 mm.
1) For outer box (red)
Length of one big box (9 big boxes are in figure)= 1mm ie =0.1cm
Breath of one big box (9 big boxes are in figure)= 1mm ie =0.1cm
Height of one big box (9 big boxes are in figure)= 0.1mm ie =0.01cm
So volume of spore suspension in one big box is =0.1X 0.1X0.01
=10 -4 mL3
10 4 = conversion factor to convert 10 -4 mL3 to 1ml
Formula
= No of cells X volume of square X DF
= No of cells X 10 -4 X DF
2) For outer box (yellow color )
Length of one box (16 boxes are in figure)=0.2mm ie =0.02cm
Breath of one box (16 boxes are in figure)= 0.2mm ie =0.02cm
Height of one box (16 boxes are in figure)= 0.1mm ie =0.01cm
So volume of spore suspension in one big box is =0.02X 0.02 X0.01
=10 -6 mL3
10 6 = conversion factor to convert 10 -6 mL3 to 1ml
Formula
= No of cells X volume of square X DF
= No of cells X 10 -6 X DF
3) For outer small box (Green color )
Length of one box (25 boxes are in figure)=0.25mm ie =0.025cm
Breath of one box (25 boxes are in figure)= 0.25mm ie =0.025cm
Height of one box (25 boxes are in figure)= 0.1mm ie =0.01cm
So volume of spore suspension in one big box is =0.025X 0.025 X0.01
=6.25X10 -6 mL3
6.25 x 10 6 = conversion factor to convert 6.25 x10 -6mL3 to 1ml
Formula
= No of cells X volume of square X DF
= No of cells X 6.25X10 -6 mL3 X DF