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The in situ staining method of Chen (1977) for the detection of mycoplasma contaminants in tissue cultures was tested in cultures of human skin fibroblasts after controlled contamination with Mycoplasma arginini. It is concluded that this method is reliable only at infection rates of 100% or higher, i.e., at one mycoplasma or more per tissue-culture cell.
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The acridine orange test for detection of Trichomonas vaginalis in smears has been adapted for delayed examination of specimens. Mailed-in slides stained by acridine orange were compared with on-site wet mounts; the acridine orange test detected 96% of all positives, whereas only 76% were detected by wet mounts. In a similar comparison with Papanicolaou smears, the acridine orange test detected 89% as compared with 67% detected by Papanicolaou smears.
Polycarbonate Nuclepore filters are better than cellulose filters for the direct counting of bacteria because they have uniform pore size and a flat surface that retains all of the bacteria on top of the filter. Although cellulose filters also retain all of the bacteria, many are trapped inside the filter where they cannot be counted. Before use, the Nuclepore filters must be dyed with irgalan black to eliminate autofluorescence. Direct counts of bacteria in lake and ocean waters are twice as high with Nuclepore filters as with cellulose filters.
Comparisons were made in Lusaka and in London between manual staining and staining in an automatic machine with auramine-phenol of direct smears of sputum and other types of specimen for acid-fast bacilli. No evidence was obtained of carry-over of acid-fast bacilli from positive to negative smears during machine staining. There was improved contrast between bacilli and the background in smears prepared with the machine.
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For fixation of the material to be investigated at the light and ultrastuctural level the author suggests to use 4% formaldehyde obtained by depolymerization of paraform on 0.1 M phosphate buffer with end pH 8.0 of the solution. This fixative stabilizes well glycogen and tissue structure. The ultrastructural characteristics of cells following continuous (up to 130 days) fixation are comparable with those obtained after fixation with aldehydes by conventional techniques. Results of histological PAS test, Feulgen's reactions and those to acid polycaccharides were identical to the results obtained in fixation with neutralized formalin. The possibility of prolonged storage of tissues makes it possible to perform delayed and repeated ultrastructural investigations of clinical and experimental material.
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Bone defects and osteotomies of the rabbit tibia were stabilized by a mixture of ethyl cyanoacrylate and bone meal. Healing of osteotomy or new bone formations growing through the defect could not be observed.