The surface area of fields of view microscope used in our study
The surface area of fields of view (SA-FOV) of the DM 200 LED microscope used in the study under various magnifications and other important parameters. The table shows diameters and surface areas of microscopic fields under various magnifications used to analyze Gram-stained samples along with some morphometric parameters important for the methodology. The entire surface area of the microscopic slide was 1935 mm2, and a third was considered as an average stained area - 1935 mm2/3 ≈ 600 mm2. Based on these data, the number of fields of view (FOV) was calculated for various magnifications. It is clear from the Table that the surface area (SA) of FOV under magnifications x400 and x200 are 6.25X, and 25X larger than SA-FOV under magnification x1000, respectively. The average number of epithelial cells (EC) per FOV was assessed by halving their maximal number given the average SA-EC (≈0.002 mm2), according to the formula (SA-FOV/SA-EC)/2. Considering the surface area of an EC as constant and roughly equal, at a FOV a certain average number of EC and PMNs may be seen, taking into account that SA-EC is about 40 times larger than the surface area of a PMN (0.002 mm2: 0.00005 mm2=40). Disparities in technical characteristics of microscopes make numbers of PMN impossible to compare Criteria used to interpret Gram-stained samples are quite diverse, in particular when attempts are being made to establish common criteria in the quantitative or semiquantitative assessment of numbers of cells and bacteria irrespective of the specimen type: vagina, sputum, cerebrospinal liquor, etc. Common to all of them is the lack of data on the area of FOV of the microscope used to establish cut-off values, this being, in our opinion, an essential part of any attempt of standardization. For example, if the HPF of our microscope is 3 times larger or smaller than of the microscope used by another research group, our cut-off values should be 2-3 times higher or lower.
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