Fig 7 (FISH guide)
LegendFig. 7. G-banding and chemical aging.a-d) Slides were chemically aged at 94 C using ethanol for 30 seconds (a), 1 minute (b), 2 minutes (c) or using dry heat for 2 minutes (d). Slides were trypsinized, then Giemsa stained. Results indicate that banding improves as the duration of the chemical aging process increases. However, G-banding is better on the dry heat aged slide compared with any of the chemically stained one. e-g)Hybridization buffer test. Three different hybridization buffers were compared by hybridizing the same cosmid probe on three slides prepared in identical conditions. Signal to noise ratio was best with the buffer used in (g). h-k)Cell suspension quality test. Same cosmid probe was hybridized on two slides from a cell suspension with more cytoplasmic residua (h, i) and on two slides from a better cell suspension (j, k). Slides were aged using chemical aging (h, j) or dry heat (i, k). Results were better with the good quality cells and were also better when using chemical aging. Arrows indicate the signals. l-n)Permeabilization test. Slides from the same sub-optimal cell suspension were kept 20 minutes at 37 C on at dry heat (l), in 4xSSC (m) or in 4xSSC/0.1% Tween 20 (n), then chemically aged and hybridized with the same centromeric probe. Slides were not enzymatically pretreated. Cytoplasmic debris are visible due to autofluorescence. Arrows in (n) point to the faint hybridization signals, visible only on the slide permeabilized with the low detergent solution.
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Last modified on: Feb12, 2001