The phase contrast microscope in plant cytology

Phase contrast microscopy affords a rapid and easy way of counting chromosomes from root-tip cells of Angiosperms. The staining method is a modified Feulgen process, the chromosomes being understained before examination under phase. This cuts down even further the time for obtaining satisfactory chromosome counts. Under phase and using a green filter the understained chromosomes in squashes, after only 30 minutes in Feulgen, appear jet black within an almost transparent cell. There is no interference from the cellulose cell walls.     

In vitro maturation of the exoerythrocytic stage of Plasmodium knowlesi observed under phase contrast microscopy

Exoerythrocytic stages of Plasmodium knowlesi were obtained in primary culture of rhesus monkey hepatocytes. The development of a single parasite was followed with phase contrast microscopy until release of merozoites in slightly less than 5 days. This direct observation may offer opportunities to determine visually factors that may influence specific steps of schizont development.     

Observations on the cytoplasmic membranes of testicular cells,examined by phase contrast and electron microscopy

In freshly isolated cells of the guinea pig germinal epithelium examined with phase contrast, dark contours are seen in the cytoplasm that appear to be optical sections of the cisternae of the endoplasmic reticulum. These increase in contrast, in number, and in linear extent with increasing time up to 4 hours after isolation of the cells from the testis. During this period, cisternae originally present in the cells are extended and new ones appear to be formed by coalescence of tubular and vesicular elements of the reticulum. The cisternae become associated in parallel array and ultimately form elaborate concentric systems resembling structures that have often been interpreted as intracellular "myelin figures." Until now our knowledge of the endoplasmic reticulum has been based largely upon electron micrographs. The observation that the cisternae are visible in certain cell types under phase contrast optics opens the way for experimental investigations on the behavior of this class of cytoplasmic membranes in living cells.     

Newly designed,simple relief phase contrast for microscopy of microorganisms

A new method providing a relief phase contrast for investigation of microorganisms by optical microscopy used a neutral filter Zeiss NG 10/1 that could be controllably slid at a certain azimuthal angle below the aperture condenser diaphragm of the microscope phase contrast. Two ways of application are described depending on the type of the microscope: (1) in a special holder, and (2) fixed on a rubber ring. The device enabled us to obtain excellent results in the area of both optical microscopy and microphotography. With the microorganisms visualized, a better resolution, higher contrast and a significant 3D effect were obtained; outer morphology and organelles (chloroplasts, nuclei, granules, oil reserve vacuoles, etc.) could also be investigated.     

Determination of Giardia muris cyst viability by differential interference contrast, phase, or brightfield microscopy

Examination of Giardia muris cysts stained with the fluorogenic dyes, fluorescein diacetate (FDA) or propidium iodide (PI), by either Nomarski differential interference contrast (DIC), phase, or brightfield (BF) microscopy revealed a direct correlation between morphologic appearance and uptake of FDA or PI. Cysts incorporating FDA were all morphologically identical and exhibited (1) a clearly delineated cyst wall, (2) the presence of a distinct space between cyst wall and cytoplasm, and (3) flagella recognizable at one pole of the cyst. FDA-positive cysts also had a hyaline appearance of the cytoplasm (examined at multiple focal planes with DIC) that made it very difficult to detect the presence of nuclei, intracellular axonemes of flagella, or curved elements of the adhesive disc. However, PI-stained cysts possessed a distinct morphology that was clearly different from that of FDA-stained cysts. Examination of PI-stained cysts demonstrated the presence of well-defined nuclei, intracellular axonemes, and curved elements of the adhesive disc. The cytoplasm of PI-stained cysts contained a fine granular texture as opposed to the hyaline appearance of FDA-stained cysts, and no space was observed separating the cyst wall from the underlying cytoplasm in the PI cyst. This light microscopic comparison of viable FDA- and nonviable PI-stained cysts of G. muris demonstrates that 2 types of cysts can be distinguished and implies that structural differences can be used to identify these subpopulations of cysts.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immediate diagnosis in vaginal cytology using interference contrast microscopy (Nomarski).

Studying fresh aspiration material from the posterior fornix and cervix, by Interference Contrast Microscopy (Nomarski) is a good method of cytologic examination. It is shown how most cell types can be observed, just as they can be by the classical Papanicolaou staining. Normal and abnormal, even dysplastic and malignant cells can be recognized. This method is also very useful for identifying parasites, fungi and bacteria, by morphology and active movements. While encoraging the use of this method, it is advisable to compare the results with slides examined later by the Papanicolaou technique, for maximum safety of the patient.     

Morphological study of the bovine blastocyst with phase contrast microscopy

The morpholoqical appearance of 40 bovine blastocysts. collected from single ovulating heifers 7 days after oestrus was evaluated with the aid of a phase contrast inverted microscope. Out of 40 blastocysts, 17 were classified normal (N), 14 in the process of degeneration (IPD) and 9 degenerated (D). There was no difference in the mean diameters of the three groups of blastocysts, whereas the mean diameters of cell mass was bigger in N than IPD and D blastocysts. This difference was highly significant (P < .005). The mean number of cells was greater in normal than in abnormal (99 vs 58). This difference was highly significant (P < .005). The results reported here indicate that the criteria used for classification of the morphological appearance of the 7 day old bovine blastocyst reflected the degree of cellular organization and the number of cells.