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The numbers of bacterial cells per milliliter were quantified by microscope cell counting in a Burke- Turk chamber (Blaubrand®) according to the manufacturer's protocol.
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The viable cell density was measured using the trypan blue (Sigma, Ronkonkoma, NY, USA) exclusion method with a hemocytometer and light microscope for manual cell counting.
Cell number was determined by manual cell counting using a microscope with a counting chamber.
For assessing in vitro proliferation, Lin−, Sca-1+, c-kit+ (LSK) stem cells were sorted from the BM into a 48 well plate (100 cells/well), and clonal expansion was followed for 4 days by manual cell counting under the microscope.
Cell growth was determined by direct cell counting under the microscope after cells were treated with various concentration of NS-398 for 24 or 48 h.
The other fraction was cytocentrifuged (Shandon, Inc., Pittsburgh, PA) onto a microscope slide and stained for differential cell counting using Diff-Quik stain (Fisher Scientific, Pittsburgh, PA).
Using a cell counting chamber placed under a phase contrast optical microscope, 64 samples from each cell type were counted.
Cell counting using a haemocytometer and cell counting using a microscope remain the reference methods.
The number of cells in each detection point was recorded under optical microscope using cell count plates, so as to draw the cellular proliferation curve.
Clones were imaged on a Zeiss 510 confocal microscope, and cell counts were taken from the two slices which represented the largest total area.
Samples were observed and photographed under a light microscope; the cell count was determined by densitometry.
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