For immunofluorescence microscopy, cells were grown on acid-treated coverslips, fixed with 4% paraformaldehyde/PBS for 15 min, and permeabilized with 0.1 % Triton X-100/PBS for 5 min. After blocking with 5 % goat serum, the coverslips were incubated with primary antibodies overnight, followed by appropriate secondary antibodies conjugated with Alexa Flour 488 and Alexa Fluor 594 Phalloidin for 30 min. The coverslips were mounted on slides using DAPI-containing mounting solution (Vector Laboratories) and observed using a Zeiss LSM 710 laser confocal microscopy.
Lsm 710 laser confocal microscopy
Manufactured by Zeiss
Sourced in Germany
The LSM 710 is a laser confocal microscopy system developed by Zeiss. It is designed to provide high-resolution imaging of samples by using a focused laser beam to scan the specimen and collect fluorescence or reflectance data. The system includes features such as adjustable pinhole sizes, multiple laser excitation options, and sensitive detectors to capture detailed information about the sample.
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Lab products found in correlation
2 protocols using lsm 710 laser confocal microscopy
1
Immunofluorescent Labeling of Cellular Structures
2
Immunofluorescence Staining of Frozen Aorta
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Frozen aorta tissue sections were fixed in 4% paraformaldehyde for 15 min at room temperature, then rinsed twice with PBS, and permeabilized with 0.2% Triton X-100 for 20 min. Blocking was done for 1 h with a blocking solution consisting of 1% BSA and 0.1% Triton X-100 in PBS. Sections were then incubated overnight with anti-RhoA primary antibody at 1:100 dilution in 1% BSA and 0.05% Tween-20, then rinsed twice with 0.1% Tween-20. A goat anti-rabbit secondary antibody, conjugated to Alexa Fluor (AF594 IgG, Invitrogen, USA), was then added at 1:250 dilution in 1% BSA and 0.05% Tween-20 for 1 h in the dark. Slides were then rinsed five times in 0.1% Tween-20 at 10 min intervals. The nuclear stain 4′,6-diamidino-2-phenylindole (DAPI) was used at 1:5000 dilution and sections were incubated for 20 min in the dark. Imaging was done using a LSM710 laser confocal microscopy (Zeiss, Germany).
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