Axiocam 105 color

Otolith imaging and shape analysis were carried out according to the procedure described in [29 (link)]. In brief, high-contrast images of entire and clean cod otoliths were taken with a stereo-microscope (SZX10, Olympus) equipped with a digital microscope camera (Axiocam 105 color, Zeiss). Subsequent shape analyses on otolith images were conducted using normalized elliptic Fourier descriptors from the ShapeR package [44 (link)] of R v4.2.1 [45 ]. The first 12 harmonics (giving 48 shape coefficients) reached 99% of cumulated power percentage and were chosen to describe the shape variations of Baltic cod otoliths.

Fresh kidney from each mouse was fixed in 4% formaldehyde and embedded in paraffin. Then, 4-μm thick sections were transferred to positive charged glass slides and processed for Hematoxylin and Eosin (H&E) staining. Images were captured using an Axio Lab. A1 microscope. Images were captured with Axiocam 105 color (ZEISS) and analyzed with ZEISS ZEN software. Photograph analysis presents data at pixels (glomerular size) or % of stained tissue (the rest of the histological parameters).

Confocal imaging of fluorescently labeled samples and brightfield imaging (i.e, whole mount in situ hybridizations) were performed using a Nikon A1 Laser Scanning Confocal Microscope or a Zeiss Axio Zoom V16 equipped with a transmitted light base and a Zeiss Axio Cam 105 Color camera, respectively.

Optical images of clean, soiled and soiled clot patches washed with 500 Gy assisted biosurfactant at 60 °C under stirring conditions were captured using AX10 Zeiss light microscope coupled with Axiocam 105 color (Germany) at NCRRT. Images representing the fabric threading were captured at 25X and 100X magnification.

Reproductive organ morphology was examined using a microscope (ZEISS, VMI 0560) equipped with differential interference contrast (DIC) optics. 4′,6-diamidino-2-phenylindole (DAPI) was used to stain nuclei, especially those of potential sperm, as follows. Young adults (N = 15) were placed into 20 μl of 100% methanol (in a 1.5-ml Eppendorf tube) and then incubated at 4 °C for 30 min. Next, filter paper was touched to the methanol liquid surface for ∼5 s to wick away most of the methanol. Then, 15 μl of 200 ng/ml DAPI was added into the tube and the worms were incubated at room temperature (approximately 25 °C) for 30 min. Filter paper was used to wick away most of the DAPI solution, and then a drop of water was added into the tube. DAPI stained worms were observed at 63× magnification with an inverted microscope (ZEISS, VMI 0560) equipped with a camera (Axiocam 105 color) and a fluorescent light source.

Between 5 and 10 high-resolution images per animal, with 5–8 animals per group, were captured by an Axiocam 105 color (Zeiss, Jenna, Germany) camera under the same parameter settings for adipocyte morphometry analysis, using a Zeiss microscope Mod. Axioplan 332 2 (Zeiss, Jenna, Germany). Slides were imaged at 10-fold magnification. To quantify the mean adipocyte area, mean adipocyte diameter, adipocyte area distribution and adipocyte diameter distribution, an average of 512 adipocytes per animal were analyzed using the FIJI (ImageJ v2) plugin Adiposoft. Images were captured and imported into ImageJ software and analyzed by the ImageJ plugin Adiposoft. To ensure that only adipocytes were counted, each image was manually inspected. The artifacts and broken adipocytes unduly analyzed and accounted by the software were recorded and manually eliminated from the output-excel sheet that contained the diameter and area data of all the adipocytes in each image. The mean adipocyte area and mean adipocyte diameter were calculated for each animal and for each group.