Reaction of Blepharisma to Golgi Impregnation Methods.

Abstract

From a study of the normal and regenerating contractile vacuoles in vivo, evidence was obtained that the normal contractile vacuole in Blepharisma undulans is not a permanent cell organ, but a system of temporary, potentially independent fluid vacuoles. 1 In view of the fact that conflicting opinions concerning the permanence of contractile vacuoles have arisen from the failure of investigators to study both living and stained preparations, the above conclusions have been tested by a study of fixed material, derived from the pedigreed cultures. The Nassanov methods and the original Kolatschev technique were employed, not only because of their proved success in demonstrating contractile vacuole structure, but also because by their use the Blepharisma vacuole could be examined in the light of the Nassanov homology.

In the several series of preparations which were made, Paramecia, mixed with the Blepharisma prior to fixation and treated identically step by step, were used as controls. Although the contractile vacuoles of the Paramecia showed the characteristic blackening, those of the Blepharisma failed to do so. In general, where osmication was continued until the whole cytoplasm was darkened, the food and contractile vacuole walls in the Blepharisma were likewise blackened, but upon differentiation of such preparations in turpentine, these walls became bleached as quickly as the surrounding cytoplasm. Particularly careful examination of the preparations showing darkened vacuolar walls for any sign of the old primary vacuole wall which, if present, should form a small dark mass close to the anal spot, revealed no trace of that structure.