Counting cells with a haemocytometer A haemocytometer is a specialized microscope slide used to count cells, organelles, etc.

It has a thick base and uses a special coverglass which is thick enough to stay flat under the pull of surface tension from the solution in the counting chamber. To load: Assemble the haemocytometer. Inject ~15 l of solution per side, using the loading notch. Make sure that the chamber is fully loaded with liquid, plus some excess in the channels beside it.

Appendix 10 Page 1 of 1

The centre portion of the slide has etched grids with precisely spaced lines. The coverslip is positioned 100 m above the slide. The shaded square (left) has sides of 200 m x 200 m, bounded by three lines. The centre line marks the 200 m spacing. The volume over the shaded square is 200 x 200 x 100 m3, or 4 x 106 m3. 1 l = 1 mm3 = 109m3. The volume over the 200m square is 4 x 10-3 l, which is one 250th of a l The smallest square has sides 50 m x 50 m. The volume over the smallest square is 2.5 x 104 m3, which is 2.5 x 10-4 l, which is one 4000th of a l. To count Aspergillus spores: make a 1:100 dilution of a harvested culture. Count the number of spores in 10, 200 m squares (five per side), and average. Calculate the number of spores per l as (average per 200m square) x 250 x 100 (dilution factor, which can vary as needed)