By Shirley McBride, Mary Heenan, Martin Clynes (auth.), Dr. Martin Clynes (eds.)
Cell tradition recommendations let numerous molecular and telephone organic inquiries to be addressed, supplying physiological stipulations while heading off using laboratory animals. as well as simple options, quite a lot of specialized useful protocols overlaying the subsequent components are incorporated: mobilephone proliferation and loss of life, in-vitro versions for mobile differentiation, in-vitro versions for toxicology and pharmacology, business program of animal cellphone tradition, genetic manipulation and research of human and animal cells in culture.
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S. Tel. 1 617 868 6200 Fax. 1 617 868 0355 Toll Free 1 800 492 1110 Nunc. A/S Kemstrupvej 90 Roskilde DK 4000 Denmark Tel. 45 42 359065 Fax. 4542 350165 2 Cell Culture and Diagnostic Virology Bibby Sterilin Ltd, Tilling Drive Stone, Staffordshire STl50SA UK Tel. +44(0) 1785-812121 Fax. +44(0) 1785-813748 BDH Laboratory Supplies Poole Dorset UK BH151 TD Tel. (01202) 660-444 Fax. (01202) 666-856 Becton Dickinson European Division Chemin des Sources BP 37 Meylan CEDEX, France 76 416464 (phone) Hycor Biomedical Inc.
1. Prepare cell cultures in flat bottomed tissue culture microtiter plates by seeding the wells with 200 fll of cell suspension containing 1 x 105 cells/ml. 2. Cover the plate and incubate at 37°C in a moist CO 2 atmosphere. 3. When the cells are confluent, replace GM with 100 fll MM. 4. Make 10-fold serial dilutions of the virus isolate in clean sterile glass containers. 5. Reconstitute the specific anti-serum to a working concentration. 6. Mix equal volumes (100 fll each) of virus and anti-serum and incubate at 37°C for 1 h.
10. Mount the coverslips onto the prelabeled slides with a drop of the mounting medium. Dry off the bottom of each coverslip carefully with lint-free tissue before mounting top side down on the slide. Place each of the preparations into a slide folder and store in the dark until ready to analyze. Preferably, the samples should be analyzed on the day of staining, but may subsequently be stored in the dark for further analysis if necessary. 3 Screening for Mycoplasma Contamination in Animal Cell Cultures 45 Fig.