Hoechst-stained 40-μm serial sections were sampled through the entire hippocampus. From a random starting position, sections at evenly spaced intervals were sampled to obtain an unbiased stereological assessment of hippocampal volume (Gundersen et al. 1988 ). In short, sections from halved brains were taken from Tat− (n = 6) and Tat+ (n = 5) mice and imaged using a Zeiss Axio-Observer Z1 microscope, MRm digital camera, and a 10× objective (Zeiss, Oberkochen, Germany). Montages of the entire left hemisphere of the brain (5 × 6) were obtained utilizing a motorized stage encoder (Zeiss) and a computerized tile-reconstruction method (Zeiss, Axio-Vision software). Hippocampal volume was then estimated by overlaying a standardized grid over the image of the brain and performing a point count analysis (Gundersen et al 1988 ; Mouton 2002 ).
Mrm digital camera
Manufactured by Zeiss
Sourced in Germany
The MRm digital camera is a high-performance, scientific-grade imaging device designed for laboratory and research applications. It features a large-format CMOS sensor, advanced image processing capabilities, and precise exposure control, enabling the capture of detailed, high-quality images for scientific analysis and documentation.
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2 protocols using mrm digital camera
1
Unbiased Stereological Assessment of Hippocampal Volume
2
Quantifying Immune Cell Populations
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Cells were plated onto sterile cover glass within 6-well culture plates for 24 h. On the day of labeling, cells were washed twice in warm PBS containing 0.5% BSA and subsequently blocked in 20% normal goat serum for 15 min in the incubator. Cells were again washed in warm PBS+BSA before application of a 1:10 dilution of anti-human CD45-FITC antibody (Miltenyi Biotec, San Diego, CA, USA) in combination with 1 μg/mL Hoechst (Thermo Fisher Scientific) for 30 min in the incubator. Cells were washed twice in PBS + BSA before immediate live cell image acquisition.
Cells were observed immediately using a Zeiss Axioplan 2 epifluorescence microscope and Axiovision software. Images were acquired using a Zeiss MRm digital camera. A minimum of 200 cells in five separate fields for each sample were assessed for Hoechst staining (total cells) and CD45-positive immunoreactivity (positive cells).
Cells were observed immediately using a Zeiss Axioplan 2 epifluorescence microscope and Axiovision software. Images were acquired using a Zeiss MRm digital camera. A minimum of 200 cells in five separate fields for each sample were assessed for Hoechst staining (total cells) and CD45-positive immunoreactivity (positive cells).
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