Interpretation of results

The simplest method is to assume that the grain observed overlies the point of radioactive decay and hence represents the site of the target molecule. For lower resolution studies at the light microscopic level this assumption may work; however, in the case of electron microscopy autoradiography this assump-

Figure 1 Binding and distribution of B3 (a costimulatory molecule) on the surface of T cells. B3 coupled to liposomes was labeled with 125l and incubated with anti-CD3-activated CD4* T cells and electron microscopic autoradiography was performed using llford nuclear L4 emulsion, (a) Negative staining of liposomes by phosphotungstic acid (bar = 200 nm); (b) 12sl-labeled B3 coupled to liposomes; arrows show the liposomes bearing the iodinated B3 (bar = 200 nm); (c) CD4' T cell labeled with B3 (arrows) (bar = 200 nm). (Reproduced from Vinay, Raje, Verma and Mishra (1995).)

Figure 1 Binding and distribution of B3 (a costimulatory molecule) on the surface of T cells. B3 coupled to liposomes was labeled with 125l and incubated with anti-CD3-activated CD4* T cells and electron microscopic autoradiography was performed using llford nuclear L4 emulsion, (a) Negative staining of liposomes by phosphotungstic acid (bar = 200 nm); (b) 12sl-labeled B3 coupled to liposomes; arrows show the liposomes bearing the iodinated B3 (bar = 200 nm); (c) CD4' T cell labeled with B3 (arrows) (bar = 200 nm). (Reproduced from Vinay, Raje, Verma and Mishra (1995).)

tion need not hold true and careful analysis of the images obtained has to be made. There are two methods in common use for localization of the radioactive decay event in a sample with relation to the observed grain density. These are known as the probability circle analysis and the cross-fire method (see Williams (1985) for detailed methodology).

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