Prefrontal tissue sections from a naive adult rhesus monkey were processed for immunohistochemistry with NeuN (Millipore, 1:1,000) and Olig2 (Abcam, 1:2,000) antibodies, in conjunction with Alexa Fluor 488 goat anti-rabbit and Alexa Fluor 594 goat anti-mouse IgGs (Invitrogen), using standard procedures. In a second, independent set of immunohistochemistry experiments, small tissue blocks (surface area <0.25 cm2) were dissected from the dorsolateral prefrontal cortex (Area9/46), immersion-fixed in 4 % paraformaldehyde solution for 15–18 hours, dehydrated in 70 %, 90 % and 100 % ethanol (3 × 30 min each) followed by xylene immersion (3 × 20 min) and paraffin-embedded using a Tissue TEK embedding center. Paraffin embedded tissue blocks were cut in 8 µm thick sections and mounted on slides for immunohistochemistry. Sections of three clozapine-exposed animals with elevated proportions of NeuN+ nuclei in white matter were de-paraffinized using xylazine and ethanol. The sections were rehydrated with water, permeabilized with 0.1 % Triton X-100, followed by NeuN staining (1:100, ABN78A4, EMD Millipore). After mounting the tissue with DAPI Fluoromount-G (0100-200, SouthernBiotech), images of NeuN+ subcortical white matter neurons were taken using a Carl Zeiss CLSM780 microscope. Image processing was done with ImageJ (NIH).
Clsm780 microscope
Manufactured by Zeiss
The CLSM780 microscope is a confocal laser scanning microscope (CLSM) designed for high-resolution imaging of biological samples. It utilizes a laser-based illumination system and advanced optical components to achieve optical sectioning and enhanced contrast. The CLSM780 is capable of capturing detailed, three-dimensional images of specimens with the ability to selectively focus on different layers or planes within the sample.
Automatically generated - may contain errors
Lab products found in correlation
Abn78a4, by Merck Group (1 mentions)
Olig2, by Abcam (1 mentions)
Alexa fluor 488 goat anti, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Neuronal nuclei (neun), by Merck Group (1 mentions)
Dapi fluoromount g, by Southern Biotech (1 mentions)
Ab83354, by Abcam (1 mentions)
Alexa fluor 594 donkey anti mouse igg h l, by Thermo Fisher Scientific (1 mentions)
Alexa 488 conjugated ctb, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Tcs sp8 confocal laser scanning microscope, by Leica (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
3 protocols using clsm780 microscope
1
Immunohistochemical Profiling of Prefrontal Cortex
2
Visualizing Antibiotic-Induced PCs in Biofilms
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The PCs formation was studied in the biofilm using CLSM. After 5× MIC of antibiotic treatment for 4 h, biofilm was rinsed with 1 ml of 0.85% NaCl to remove planktonic cells and antibiotic. Further, the biofilm was stained with an RSG (1 µl) and PI (1 µl 1:100 diluted) dyes. To remove excess stain from the biofilm a gentle wash with 1 ml of 0.85% NaCl was given. The untreated biofilm was used as a control. The biofilm visualization was done using Carl Zeiss CLSM 780 microscope.
3
Immunofluorescence Microscopy of Measles Virus Phosphoprotein
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BGC-823 and SGC-7901 cell lines were cultured in 24-well plates at 4 × 105 cells per well. The cells after indicated treatment were gently washed with PBS, fixed in 4% paraformaldehyde (PFA) for 30 min, permeabilized by 0.1% TritonX-100, and blocked by 2% BSA. Cells were incubated with mouse anti-measles phosphoprotein (MV-P, abcam, ab43820), followed by Alexa Fluor® 594 donkey anti-mouse IgG (H + L) (Invitrogen, A-21203) and DAPI (Beyotime, C1005) in the dark, images were captured using Zeiss cLSM780 microscope. For co-localization analysis, cells were stained with Alexa 488-conjugated CTB (Invitrogen, C-34775) for 2 h at 37 °C incubator before fixation by 4% PFA. Then cells were incubated with anti-Acid sphingomyelinase (ASMase, abcam, ab83354) at 4 °C overnight, followed by Alexa Fluor® 594 goat anti-rabbit IgG (H + L) (Invitrogen, A-11012) and DAPI in the dark. Images were captured using Lecia TCS SP8 confocal laser scanning microscope.
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