Sensitmty of Secondary Response in Vitro
نویسنده
چکیده
Antigens, preparation of lymph nodes and culture media, stimulation in vitro, implantation in clotted plasma, incubation, and estimation of antibody t.iters on fluid withdrawn at 3 day intervals, were similar to those in the accompanying paper (1). In most of these experiments, the fragments were exposed to antigen by incubation before implantation into plasma. Histological Methods for Fluorescent Studies.--To facilitate microscopic examination in situ, we occasionally implanted the fragments on a coverslip attached to the wall of the tube by capillarity. In addition, fragments were removed from the tubes at intervals, fixed in 95 per cent ethanol, and paraffin-embedded by the method of Sainte-Marie (2). Fluoresceinconjugated hyperimmune rabbit anti-bovine serum albumin (BSA) and diphtheria (D) antitoxin were purified by absorption with mouse liver powder, or by DEAE cellulose column chromatography by the method of McDevitt a al. (3). They were used for the histological localization of the anti-BSA and anti-D (4). Control slides were prepared by omitting the intervening antigen layer. Bouin-HoUande-fixed, paraffin-embedded Domlnlci-stained sections were used for routine histological examinations (2).
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