Description and analysis of a simple micro-titration immune cytolytic test

Abstract

A method of micro-titration for cytolytic antibodies is described. Standard drops of diluent are placed on a subdivided siliconed slide and dilutions are made by rotating a wire loop holding 1 drop of serum in each drop sequentially. One drop of a mixture of cells and complement is then added. The slide is incubated in a moist chamber for 90 min at 37° C. after which a small cover-slip is placed on each drop and the amount of cell lysis, judged by phase-contrast microscopy, is graded.An analysis of variance of the method has been performed and the method has been found to be reliable, accurate and unbiased.It allows accurate estimates of cytolytic activity in sera using cell concentrations of 104 to 105 cells per ml. Eight ten-dilution titrations can be carried out with 1 ml. of such a suspension.

Comparative Studies on the Rates of Re‐Expression of Human Histocompatibility Antigens after Papain Treatment

“…The range is quite wide and was observed to be particularly wide between experiments performed on different days. The main fartor accounting for this is probably day to day variation in experimental technique, as described by Spooner et al (1965).…”

Antibody Plaque Formation <i>in vitro </i>in the Presence of Respiration Inhibitors

“…The direct effect of inhibitor on the serological reactivity of hemolysins was determined by a modification of the method of Spooner et al [6]. In brief, pooled mouse or rabbit immune serum containing high titered hemolysins was diluted in microtiter plates, using 0.025 ml buffered saline as the diluent.…”

Effect of Hydrocortisone and ε-Amino Caproic Acid on Immune Cytolysis in vitro

“…R. Batchelor and were subsequently grown in C3H/Hef mice. The cells were stored in a liquid nitrogen container with methyl cellulose added to 1.3% and dimethyl sulphoxide to 13% (Spooner, 1965). The cell suspensions were thawed rapidly, resuspended in Hanks's BSS followed by one wash in Hanks's BSS.…”

“…The technical procedure was that described by Spooner, Bowden and Carpenter (1965). Briefly, complement (or antiserum) dilutions were made with Takatsy-typc microtitration loops in 0.025 ml volumes on a subdivided siliconed slide.…”