CSF sample should be properly mixed. If CSF is clear, it is not diluted. If CSF appears cloudy or turbid, 1:20 dilution is made using 0.05 ml of CSF and 0.95 ml of Turk solution. (Composition of Turk solution: Glacial acetic acid 4 ml, methylene blue solution 10 drops, and distilled water to make 200 ml). The counting chamber is covered with the coverslip provided. The counting chamber is filled with the fluid and allowed to stand for 2 minutes for cells to settle. iv. For counting cells in CSF, cells are counted in 5 large squares (4 corner squares and one central square). v. If undiluted CSF is used, total number of cells counted in 5 squares represents total count per cmm of CSF. If CSF is diluted, the number of cells counted is multiplied by the dilution factor (i.e. 20).