Cells were cultured and transfected on Lab-Tek II chamber slides. Cells were fixed for 15 min in 4% formaldehyde, washed with phosphate buffered saline (PBS) 3 times, and incubated in blocking buffer (PBS, 5% goat serum, 0.1% saponin) for 1h at room temperature. Primary antibody was diluted in antibody solution (PBS, 1% BSA, 0.1% saponin) and incubated at 4C overnight. After washed with PBS 3 times, cells were incubated with secondary antibody conjugated with Alexa Fluor 488 or 568 (Invitrogen) for 1h at room temperature. After PBS wash, the slides were mounted with Prolong Gold Antifade reagent. The immunofluorescent pictures were taken by the Zeiss LSM510 Meta Confocal Laser Scanning Microscope or Nikon Eclipse E800, and analyzed by the LSM image browser or Nikon NIS Elements BR 3.0 software.
Lsm510 meta confocal laser scanning microscope
Manufactured by Nikon
The LSM510 Meta Confocal Laser Scanning Microscope is a high-performance research-grade microscope system designed for advanced imaging applications. It utilizes laser excitation and confocal optics to capture detailed, high-resolution images of biological samples. The system is capable of multi-channel fluorescence imaging, optical sectioning, and 3D reconstruction.
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Lab products found in correlation
Eclipse 80i microscope, by Nikon (2 mentions)
Alexa fluor 488 or 568, by Thermo Fisher Scientific (1 mentions)
Nis elements br 3, by Nikon (1 mentions)
Eclipse e800, by Nikon (1 mentions)
Fv1000 confocal microscope, by Zeiss (1 mentions)
Lsm image examiner, by Zeiss (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Fv10 asw, by Olympus (1 mentions)
2 protocols using lsm510 meta confocal laser scanning microscope
1
Immunofluorescence Microscopy of Cells
2
Microscopic Imaging of Filamentous Cyanobacteria
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Microscopic studies and photographs were carried out using Olympus FV1000 confocal microscope, ZEISS LSM 510 META confocal laser scanning microscope, or Nikon Eclipse 80i microscope (details were provided in related figure legends). DNA was stained by 4, 6-Diamidino-2-phenylindole (DAPI) (Sigma) at 1 mg/mL for 20 min before observation. Images were processed using Olympus Fluoview or Zeiss LSM Image Examiner. 3D analysis was accomplished in Olympus FV10-ASW Version 03.01.01.09. In time-lapse microscopy, filaments were cultured in a chamber slide filled with BG11 medium containing 0.5% agarose and imaged using Nikon Eclipse 80i microscope.
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