Monolayers were grown in Ibidi μdishes (Thistle Scientific) or Lab-Tek II chamber slides (Thermo) and allowed to proliferate for 24 h. Cells were treated as stated in the main text, washed with PBS and fixed in paraformaldehyde (4%, 10 mins). For samples co-stained with DAPI (500 nM, 2 min) this was added after fixation and membrane-permeabilisation (0.1% Triton, 10 mins) steps. Epi-fluorescent images were obtained using a Nikon 90i upright widefield microscope and a x60 oil-immersion objective. An in-house filter cube comprising FITC excitation with TRITC emission was employed to detect MLCT emission of 1 and 2 . Confocal images were obtained using a Zeiss LSM 780 META inverted confocal microscope and x40 or x63 oil-immersion objectives. Microscopy images were processed using either Zeiss LSM Image Browser or ImageJ software.
Lsm780 meta inverted confocal microscope
Manufactured by Zeiss
The LSM780 Meta inverted confocal microscope is a high-performance imaging system designed for advanced microscopy applications. It provides efficient optical sectioning and high-resolution imaging capabilities. The system features a flexible and modular configuration to accommodate a variety of sample types and research requirements.
Automatically generated - may contain errors
Lab products found in correlation
Lab tek 2 chamber slide, by Thermo Fisher Scientific (1 mentions)
90i upright widefield microscope, by Nikon (1 mentions)
Fluorescein isothiocyanate (fitc), by Jackson ImmunoResearch (1 mentions)
Gata4, by Abcam (1 mentions)
Ssea 1, by Merck Group (1 mentions)
Blimp 1, by Abcam (1 mentions)
Stra8, by Abcam (1 mentions)
Nanog, by Merck Group (1 mentions)
Cyanine3 (cy3), by Jackson ImmunoResearch (1 mentions)
Gh2ax, by Abcam (1 mentions)
Sycp1, by Abcam (1 mentions)
Elyra ps 1 microscope system, by Zeiss (1 mentions)
Fitc cy3 cy5, by Jackson ImmunoResearch (1 mentions)
Sycp3, by Abcam (1 mentions)
Rad51, by Santa Cruz Biotechnology (1 mentions)
3 protocols using lsm780 meta inverted confocal microscope
1
Fluorescence Imaging of Monolayer Cells
2
Immunofluorescence Profiling of Germ Cell Markers
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Cells or seminiferous tubules were fixed for 15 min with 4% paraformaldehyde at room temperature, blocked for 30 min with 0.3% Triton X-100/2% BSA in PBS, and incubated with primary antibodies against OCT4 (Santa Cruz Biotechnology), NANOG (Millipore), SSEA1 (Millipore), DDX4 (Abcam), BLIMP1 (Abcam), STRA8 (Abcam), and GATA4 (Abcam). After overnight incubation at 4 C, samples were washed three times in PBS, followed by incubation with secondary antibodies or/and peanut agglutinin (PNA) (10 mg/ml, Sigma) for 1 hr. Secondary antibodies were labeled with fluorescein isothiocyanate (FITC), Cy3, and Cy5 (Jackson ImmunoResearch). DNA was counterstained with 10 mg/ml Hoechst 33342 for 15 min, followed by three washes with PBS. Images were captured with a Zeiss LSM780 Meta inverted confocal microscope.
3
Immunofluorescence Analysis of Meiotic Proteins
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Cultured cells were digested into single-cell suspensions. Chromosomal spreads were prepared using a hypotonic bursting technique (Peters et al., 1997) . Primary antibodies were Sycp3 (Abcam), Sycp1 (Abcam), gH2AX (Abcam), Rad 51 (Santa Cruz), and Spo11 (provided by Scott Keeney) (Lange et al., 2011) . Secondary antibodies were FITC-, Cy3-, Cy5-, and DyLight 405labeled (Jackson ImmunoResearch). Images were captured with Zeiss LSM780 Meta inverted confocal microscope. Super-resolution analysis was performed using a Zeiss Elyra PS.1 microscope system.
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