The autoradiographic emulsion was developed, sections were counterstained with toluidine blue (VWR) for Nissl substance and mounted for visualisation using bright field microscopy for the counterstain and epi-polarized illumination for the silver grains of the autoradiographic emulsion.
Then, sections were examined with a Leica DMR microscope (Leica Microsystems, Wetzlar, Germany) using a bright field illumination for morphology and Nomarski interference contrast for immunohistochemistry. Immunohistochemical staining for both antigens, Cx43 and AR, was evaluated qualitatively in at least 20 serial sections of epididymides from each experimental group.
For comparison, Fig. 3e) shows the bright field image for LED illumination, slightly defocused to make the transparent structures visible.
Zygospores from twelve sections were counted by optical microscopy (bright field 10X) for each interaction.
Using bright field images for each 1-min interval, the projected area of the cell was manually traced (1 min before starting shear, 10 min of shear by Plasma-Lyte, 10 more minutes after flow stopped).
Once dried, the nanoparticle spots were visible in optical, MS, and electron microscope images and could therefore be used to register spatial locations across these distinct imaging modes (the CRM microscope was operated in bright field mode for sample navigation and visualization of microspots).
Cell classifier was set on area-lower limit on bright field channel (25 for PBMCs and 50 for monocyte derived macrophages and cell lines) to avoid acquiring debris.
The measurements are simultaneously carried out in two independent detection schemes viz., the bright field detection configuration for detecting the core modes and dark field detection configuration, for detecting the cladding modes.
Figure 3a, and b show the bright field TEM micrograph for the sample treated at 850°C and 950°C, respectively.
Each Diff-Quik stained sperm slide was analyzed using oil immersion (1000x) bright field light microscopy for normal morphology based on the Tygerberg strict criteria method [ 31].
