The treated cells were fixed with 4% paraformaldehyde on glass slides at room temperature for 30 min, washed with PBS, and permeabilized with 1% Triton X-100 at room temperature for 1 min. After washing with PBS, the slides were stained with an anti-TFEB antibody at 4 °C overnight. Subsequently, the slides were incubated with an FITC-conjugated secondary antibody for 1 h, washed with PBS, and mounted on coverslips with ProLong™ Gold Antifade Mountant with DAPI (Thermo Fisher Scientific). Nrf2 localization was determined using an LSM 510 multiphoton confocal microscope (Carl Zeiss, Jena, Germany).
Lsm 510 multiphoton confocal microscope
Manufactured by Zeiss
Sourced in Germany
The LSM 510 multiphoton confocal microscope is a high-performance imaging system designed for advanced fluorescence microscopy. It utilizes multiphoton excitation and confocal technology to capture high-resolution, three-dimensional images of biological samples with improved depth penetration and reduced phototoxicity compared to traditional confocal microscopes.
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Lab products found in correlation
2 protocols using lsm 510 multiphoton confocal microscope
1
Immunofluorescence Staining for TFEB
2
Immunofluorescent detection of Nrf2
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The treated cells were fixed with 4% paraformaldehyde on glass slides for 30 min at room temperature, washed with PBS, and permeabilized with 1% Triton X-100 for 1 min at room temperature. After washing with PBS, the slides were stained with anti-Nrf2 antibody overnight at 4 °C. The slides were subsequently incubated with an FITC-conjugated secondary antibody for 1 h, washed with PBS, and mounted on coverslips with ProLong™ Gold antifade mountant with DAPI (Molecular Probes, Eugene, OR, USA). The localization of Nrf2 was determined under LSM 510 multiphoton confocal microscope (Zeiss).
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