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Calcofluor white and Congo red inhibit chitin microfibril assembly of Poterioochromonas: evidence for a gap between polymerization and microfibril formation.

The influence of the light microscopical stains, Calcofluor white and Congo red, on the process of chitin microfibril formation of the chrysoflagellate alga Poterioochromonas stipitata was studied with light and electron microscopy. There is a concentration-dependent inhibition of lorica formation with both dyes. In the presence of the inhibitors malformed loricae are made, which do not show the usual ultrastructure and arrangement of the chitin microfibrils. Instead of long, laterally associated microfibrils, short rods or irregular networks of subelementary (15-25 A) fibrils are found. Microfibril assembly obviously takes place on the accessible outside of the plasma membrane. There must be a gap between the polymerization and microfibril formation reactions, allowing the stains to bind to the polymerized subunits. Thus, later association of these units to form microfibrils is disturbed. The microfibril-orienting mechanism also depends on normal microfibril formation. A model summarizing these hypotheses is suggested.

Cell Compartmentation↗

Thionine-Feulgen-Congo red staining of cervical smears for the BioPEPR image-analysis system.

A staining method developed for use with BioPEPR (Biological Precision Encoding and Pattern Recognition), an automated image-analysis system for cervical smears, is described. The stain is a combination of the Feulgen procedure, with thionine-SO2 as the Schiff reagent, and Congo red, which is used as a counterstain. The stain resulted in smears suitable both for microscopic diagnosis and for BioPEPR measurements made on photonegatives at a single wavelength 545 nm. A high level of reproducibility and accuracy of nuclear and cytoplasmic area measurements was obtained. Nuclear integrated optical density could be well measured and was shown to be useful in discriminating between normal and abnormal cells. Using a combination of morphologic features, a high level of cell classification accuracy was reached. The possibility of using the stain for more detailed studies is discussed.

Autoanalysis↗