All infected cells were visualized under fluorescent microscopy. All samples were evaluated with an inverted microscope (Nikon Eclipse TE300, Nikon, Tokyo, Japan) using phase-contrast and fluorescence microscopy. A GFP emission filter was used to detect green fluorescence, and images were obtained using NIS software. Infected tumorspheres were further examined with a Leica TCS AOBS SP2 spectral confocal microscope on an upright stand (Leica Microsystems, Wetzlar, Germany) following staining with Annexin V- Cy5 Apoptosis Detection Kit (BioVision, Mountain View, CA) and 4′, 6-diamidino-2-phenylindole dihydrochloride DAPI for measures of early and late apoptosis. Time-lapse images of sphere formation and cellular infection were obtained using a Zeiss LSM 5 Live confocal laser scanning microscope (Carl Zeiss, Oberkochen, Germany).
Lsm 5 live confocal laser scanning microscope
Manufactured by Zeiss
Sourced in Germany
The LSM 5 Live is a confocal laser scanning microscope manufactured by Zeiss. It is designed to provide high-speed, real-time imaging of live samples. The microscope uses laser technology to illuminate and scan the specimen, capturing detailed images with improved resolution and contrast compared to traditional optical microscopes.
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Lab products found in correlation
2 protocols using lsm 5 live confocal laser scanning microscope
1
Imaging of Infected Tumorspheres
2
Immunofluorescent Localization of NLRP3 in PCa Cells
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The human PCa cell lines (PC3 and LNCaP) were fixed with 4% paraformaldehyde for 30 min, permeabilized with 0.5% Triton X-100 for 15 min, and incubated with 5% bovine serum albumin (BSA) in PBS for 1 h at room temperature. Subsequently, cells were incubated with primary antibodies against NLRP3 (1:200, Abcam, London, UK) overnight at 4 °C. After three washes with PBS (5 min per wash), cells were incubated with anti-rabbit (1:500, Invitrogen) for 1 h at room temperature. After three washes with PBS, nuclei were stained with Hoechst 33342 (Beyotime, Nantong, China) for 30 min. Immunofluorescence images were captured using a Zeiss LSM5 Live confocal laser scanning microscope (Carl Zeiss, Jena, Germany).
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