Computer analysis of living cells: movements of the chromatoid body in early spermatids compared with its ultrastructure in snap-frozen preparations

Abstract

 Quantitative analyses of cytoplasmic and nuclear organelle movements in living interphase cells at defined stages of differentiation are few. By phase contrast videomicroscopy and digital imaging techniques, we have traced the path of the chromatoid body (CB) and analysed its rapidly changing positions in relation to the nuclear envelope, Golgi complex and nuclear pale chromatin areas in living early spermatids of the rat. The CB had intimate interactions with the nuclear envelope and moved both in parallel and perpendicular fashion in relation to it. It had successive short contacts with the Golgi complex and nuclear pale chromatin areas. It was also seen to scan between two pale chromatin areas and it had pinocytosis-like transient engulfments during interactions with the pale chromatin. In ultrastructural analysis of snap-frozen preparations, the CB had a large contact area with the nuclear envelope with several intermediate organelles that may be involved in nucleocytoplasmic material transport. It is evident that quantitative image analysis of living cells is a powerful guide for ultrastructural analyses. The snap-freezing technique gives new possibilities for studies of structures that are sensitive to conventional fixation procedures.