Assessment of the viability of Schistosoma mansoni schistosomula by comparative uptake of various vital dyes

Abstract

Of various vital dyes used to assess schistosomula viability, toluidine blue enabled differential counting of the schistosomula on microscope slides but not in culture wells, whereas methylene blue could be added directly to the schistosomula suspension in culture wells of microtiter plates. Toluidine blue uptake by dead parasites was very fast. It mostly also partially stained damaged but not dead organisms. Its main disadvantage was rapid, nonspecific staining of live schistosomula, requiring prompt counting of a preparation and additional reliance on motility for assessment of viability. Methylene blue staining of dead worms was slower, but it did not stain the live worms until about 1 h after dye application, enabling its addition to a series of preparations for consecutive counting. It did not always stain flattened, dead schistosomula or it stained them an uncontrasting pale blue. This dye remarkably induced movement in seemingly inert and probably damaged worms, thus enabling determination of viability even following poor staining or a lack of staining.