By John Swansbury
A set of key cytogenetic and FISH recommendations utilized by smooth scientific laboratories within the genetic research of human malignancies. The book's sensible recommendation and techniques are appropriate to be used at each point of workmanship, together with absolutely validated laboratories, yet with a sympathetic bias in the direction of somebody contemplating developing a brand new cytogenetics provider. right here the reader will locate not just ordinary tutorials at the basics of human karyotypes and chromosome research, but additionally unique discussions on how laboratories may possibly optimally improve their repertoire of features to incorporate such more moderen complementary suggestions as CGH, FISH, and M-FISH.
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Additional info for Cancer Cytogenetics. Methods and Protocols
B. Using dry slides: Label the dry slides and place flat on a slide tray. Add two drops of cell suspension, then immediately blow gently, or “huff,” on the slides. c. Using wet slides: Label the dry slides. Dip slides singly or in pairs into a beaker of distilled water; ensure that there is an even film of water on each slide. Add two drops of cell suspension to each slide, then gently flick off the excess water. Stand upright to let the remaining water drain away. 4. Leave the slides to dry at room temperature.
The slides must be free of dust and grease. Specially cleaned slides may be bought, otherwise wash in detergent, rinse well in water, then in dilute hydrochloric acid and alcohol. 14. Stains: Wright’s stain. ) This is usually obtained as a powder. Cover a flask with aluminum foil, and insert a magnetic stirrer. 5 g of stain and 200 mL of methanol. Stir for 30 min. Filter through filter paper into a foil-coated bottle. Close the lid tightly, and store the bottle in a dark cupboard for at least a week before use.
In the author’s laboratory, it is the policy that as far as possible samples are harvested at the normal times, even if it means staff coming in to work on a day when the laboratory is usually closed. An alternative is to set up the cultures and then place them in a refrigerator at 4°C; on the day before the laboratory reopens, a member of staff calls in to move the cultures from the refrigerator to the incubator. 56 Swansbury 3. Previously published protocols for blocked cultures recommend adding the releasing agent next morning, then leaving the cultures for some hours before adding colcemid, with harvesting starting a short time (10–20 min) later.