Cells were fixed with 4% PFA in PBS for 15 min at 37°C, then washed twice with PBS, permeabilized with 0.1% Triton X-100 in PBS for 10 min, washed four times with PBS, and blocked with 3% bovine serum albumin in PBS, all at room temperature. For double staining, the cells were incubated with appropriate primary antibodies for 1 h at room temperature, washed three times with PBS, and then incubated with appropriate secondary antibodies for 30 min. The samples were washed as before, mounted using Fluorescent Mounting Medium (Dako), and analyzed using an Olympus FV1000-D confocal fluorescence microscope.
Fv1000 d confocal fluorescence microscope
Manufactured by Olympus
The FV1000-D confocal fluorescence microscope is a high-performance imaging system designed for biological research. It utilizes laser-scanning technology to capture detailed, high-resolution images of fluorescently labeled specimens. The microscope is capable of imaging multiple fluorescent probes simultaneously, enabling the visualization of complex cellular and subcellular structures.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using fv1000 d confocal fluorescence microscope
1
Immunofluorescence Staining Protocol
2
Fluorescent Staining and Microscopy Analysis of Mouse Brain Slices
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were perfused with 4% paraformaldehyde (PFA) and the brains were isolated. Brains were fixed in 4% PFA overnight at 4 °C, and 40 μm slice sections were prepared. For fluorescent stains, sections were incubated in 0.1% Triton X-100/PBS for 15 min, washed with PBS, blocked with 0.5% blocking reagent (Roche) in 0.1% Triton X-100/PBS, and incubated for two nights at 4 °C with primary antibodies. Slices were incubated overnight with Alexa-conjugated secondary antibodies and counterstained with Hoechst 33258. For BrdU staining, sections were incubated in 0.4% Triton X-100/PBS for 10 min, washed with PBS, treated with 2 M HCl for 25 min at 37 °C, neutralized with 40 mM boric acid, washed with PBS, blocked with 0.5% blocking reagent (Roche) in 0.1% Triton X-100/PBS, and incubated for two nights at 4 °C with primary antibodies. Slices were incubated overnight with Alexa-conjugated secondary antibodies. The samples were analyzed using an Olympus FV1000-D confocal fluorescence microscope. The fluorescent area was manually measured in the ROI (0.0625 mm2) of the DG using Fiji software (37 (link)). Calretinin positive cells were manually counted in the ROI (0.24 mm2) of the DG.
3
Immunofluorescence Analysis of Brain Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Brains were fixed in 4% PFA, and 40 μm slice sections were prepared. Slices were incubated for 2 nights at 4 °C with primary antibodies. Slices were incubated overnight with Alexa-conjugated secondary antibodies and counterstained with Hoechst 33258. The samples were analyzed using an Olympus FV1000-D confocal fluorescence microscope.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!