Staining of smears

1 Place in 70% ethanol: 30min.

2 Transfer to two changes of 70% ethanol: 5min each.

3 Wash under a gently running tap: 10min.

4 Place in working solution of haematoxylin: 5min.

5 Wash in running tap water: 10min.

6 Dip slide in acid-alcohol for a few seconds.

7 Wash in running tap water: 10min.

8 Transfer to two changes of 70% ethanol: 5min each.

9 Transfer to 95% ethanol: 5min.

10 Place in absolute ethanol: 5min.

11 Transfer the slide to Histoclear: 3min.

12 Mount under a coverslip with DPX.

Examine under the light microscope using 10*, 40*, and 100* objectives for at least 100 fields. Measure objects with the calibrated eyepiece micrometer. Nuclear chromatin and chromidial bars are black, other structures, varying shades of gray to black.

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