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Fig 3 (FISH guide)
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LegendFig. 3.Comparison of reagents used for chemical aging.a-f). Giemsa staining of nondenatured, non-pretreated slides, aged 3 minutes at 94 C in: a) methanol; b) ethanol; c) 3:1 fixative; d) 1% formaldehyde; e) dry heat; f) dry heat and pepsin pretreatment. A banding pattern can be seen on all chromosomes, with few if any quality differences. Enzyme pretreatment of the heat-aged slide (3f) makes chromosomes/bands fuzzy. g-k) Giemsa staining of denatured slides, aged in: g) methanol; h) ethanol; i) 3:1 fixative; j) 1% formaldehyde; and k) dry heat. Chromosomes in 3k have a ghost-like structure. Chromosomes in 3j showed banding but, when subsequently tested, both dry heat and formaldehyde treatment decreased sharply the quality of DNA hybridization and were abandoned. Chromosomes in 3g, 3h, 3i display a similar banding quality, with chromosomes in 3i having a wider diameter. Based on this test, ethanol, methanol and fixative were further tested for was chosen for use in chemical aging.
