Use the procedure described in Subheading 3.3. to stain tissue squashes (e.g., salivary gland chromosomes), fixed in methanol/acetic acid (3 : 1 v/v) and squashed in 45% acetic or lactic acid. Use also for air-dried droplets of hemolymph, oenocytes, or adult fat body cells. To stabilize the latter types of cells, process them with their reference standards for 15 min in methanol/ formalin/acetic acid (MFA) (85 : 5 : 1, v/v), followed by rinsing in several changes of deionized water for at least 5 min before placing the slides into 5 N HCl. Deparaffinize sectioned tissues in xylenes and bring them through graded ethanols to deionized water before putting them into acid for DNA hydrolysis (see refs. 46,50,51,61 and Note 6).
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