Brain sections were collected referred in previous studies [13 (link)]. Briefly, mice were anaesthetized and transcardially perfused with 0.9% saline followed by 4% paraformaldehyde solution. The brains were removed and post-fixed over-night in 4% paraformaldehyde before being sectioned into 50 µm slices, and slices were stained with DAPI. For staining the caspase-3, fixed slices were immunostained with the caspase-3 antibody (1:500, Cell Signaling, #9661) and amplified with the goat anti-rabbit secondary antibody (1:500, Jackson, #611-605-215), and slices were stained with DAPI. Slices were imaged using the Leica TCS SP8 confocal microscope or the Olympus VS 120 slide scanning system.
Tcs sp8 confocal microscope
Manufactured by Olympus
Sourced in Japan
The Olympus TCS SP8 is a high-performance confocal microscope designed for advanced imaging applications. It features a spectral detection system that allows for the simultaneous acquisition of multiple fluorescent signals. The TCS SP8 provides high-resolution, high-sensitivity imaging with a flexible and customizable optical configuration.
Automatically generated - may contain errors
Lab products found in correlation
Vt1000s vibratome, by Leica (3 mentions)
Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (3 mentions)
Vectashield with dapi, by Vector Laboratories (3 mentions)
Vs120 microscope, by Olympus (3 mentions)
0.25na objective, by Olympus (3 mentions)
1.3 na objective, by Olympus (2 mentions)
Rabbit anti rfp, by Rockland Immunochemicals (2 mentions)
Caspase 3 antibody, by Cell Signaling Technology (1 mentions)
Goat anti rabbit secondary antibody, by Jackson ImmunoResearch (1 mentions)
Vs120 slide scanning system, by Olympus (1 mentions)
Cell f software, by Olympus (1 mentions)
Dm il led microscope, by Leica (1 mentions)
6 protocols using tcs sp8 confocal microscope
1
Immunohistochemical Analysis of Mouse Brain
2
Microscopy Protocols for Bioimaging
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Images were acquired using a Leica TCS SP8 confocal microscope or an Olympus 1 × 73 microscope with Cell^F software (Olympus Corporation). Phase-contrast images were taken on a Leica DM IL LED microscope with a Leica MC170 H9 camera (Leica Microsystems GmbH).
3
Immunofluorescence Imaging of Mouse Brain
4
Visualizing Autophagic Flux in Microglia
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To detect autophagic flux, microglial cells pretreating with TNF-α were transfected with an mRFP-GFP-LC3B adenovirus (Hanbio Co. Ltd., Shanghai, China) with or without 10 nM AT-RvD1. The fluorescence images were observed and staining was performed using the Leica Microsystems TCS SP8 confocal microscope (Olympus Fluoview™; FV1000, Japan). The colocalized red and green fluorescence puncta were calculated to show the activation of autophagy.
5
Immunohistochemical Analysis of Tdtomato Labeling
6
Immunohistochemical Analysis of Tdtomato Labeling
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Mice were anesthetized and perfused intracardially with 0.01 M PBS followed by 4% PFA. Brains were stored in 4% PFA for 12 – 18 hours at 4°C before being washed three times in 0.01 M PBS. Slices were cut on a VT-1000S vibratome (Leica) at 70 μm thickness, or 40 μm for immunostaining, and were either mounted directly onto gel-coated glass slides or processed for immunolabeling. For tdTomato immunolabeling, slices were incubated for 1 hour in blocking solution (1% bovine serum albumin and 0.2% Triton-X in 0.01 M PBS), primary antibody was applied overnight (rabbit anti-RFP (Rockland) at 1:1000 in blocking solution). Slices were then washed in PBS and incubated with secondary antibody (goat anti-rabbit 594 (abcam) at 1:400), in blocking solution) for 1.5 hours. Slices were coverslipped using ProLong Gold anti-fade reagent with DAPI (Invitrogen) or VectaShield with DAPI (Vector Labs). Fluorescent images were taken on an Olympus VS120 microscope, using a 10X 0.25NA objective (Olympus) or a Leica TCS SP8 confocal microscope, using a 20X 0.75NA and 40X 1.3NA objective (Olympus). All images were processed using NIH ImageJ.
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