Cells grown on Lab-Tek four-well glass chamber slides (NUNC) were incubated in HBSS or medium containing appropriate reagents for the indicated times. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.2% Triton X-100 for 5 min. They were washed with PBS and incubated with primary antibodies and subsequently with secondary antibody conjugates (Alexa Fluor 594 donkey anti-mouse IgG and/or Alexa Fluor 488 donkey anti-rabbit IgG; Invitrogen). Images were collected using a laser scanning confocal microscope LSM710 (Carl Zeiss, Oberkochen, Germany) equipped with argon (488 nm) and krypton (568 nm) lasers, using an x40 water immersion objective. Images were processed with ZEN 2009 light edition (Carl Zeiss).
Four well glass chamber slides
Manufactured by Thermo Fisher Scientific
Sourced in United States
The four-well glass chamber slides provide a convenient platform for researchers to conduct cell culture and microscopy experiments. Each slide features four individual wells made of glass, allowing for multiple samples or conditions to be observed simultaneously.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 710, by Zeiss (6 mentions)
Zen 2009 light edition, by Zeiss (3 mentions)
Dapi containing mounting medium, by Vector Laboratories (2 mentions)
Eclipse 90i fluorescence microscope, by Nikon (2 mentions)
Alexa fluor 594 donkey anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Ix71 inverted system microscope, by Olympus (1 mentions)
Antibodies against nrf2, by Santa Cruz Biotechnology (1 mentions)
Confocal laser scanning microscope lsm700, by Zeiss (1 mentions)
4 6 diamidino 2 phenyindole dapi, by Merck Group (1 mentions)
Fluorescein isothiocyanate fitc conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Vectashield antifade mounting medium with dapi, by Vector Laboratories (1 mentions)
Vectashield antifade mounting medium, by Vector Laboratories (1 mentions)
7 protocols using four well glass chamber slides
1
Confocal Imaging of Immunolabeled Cells
2
Visualization of BEND5 Protein Localization
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The cells were seeded in four-well glass chamber slides (Nunc, Bedford, Massachusetts, USA). After transfection for 24 h, the cells were fixed in 4% formaldehyde and stained with 4’,6-diamidino-2-phenylindole (DAPI) and anti-BEND5 antibody (1:1000, Sigma-Aldrich, SAB2700049, Taiwan, ROC). Images were captured using the Olympus IX71 Inverted Microscope System (Olympus America, Center Valley, PA, USA).
3
Immunofluorescence Analysis of Nrf2 in hMSCs
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Ethanol- or TCS-treated hMSCs were seeded at 2,000 cells per square centimeter on four-well glass chamber slides (Nalge Nunc International, Rochester, NY, USA), and the cells were incubated in a 5% CO2 incubator at 37 °C. The cells were fixed with 4% paraformaldehyde (Sigma) for 30 minutes, and then permeabilized with 1% Triton X-100 for 10 minutes followed by blocking for 1 hour with 5% bovine serum albumin (BSA) in PBS. The cells were incubated with 1:100 dilution of antibodies against Nrf2 (Santa Cruz Biotechnology) for 4 hours at room temperature, and then incubated fluorescein isothiocyanate (FITC)-conjugated secondary antibodies (Santa Cruz Biotechnology) at a 1:5,000 dilution in 1% BSA-containing PBS for 1 hour at room temperature in the dark. The nuclei were stained with 4,6-diamidino-2-phenyindole (DAPI) (Sigma) and then examined using a Zeiss LSM700 scanning laser confocal microscope (Zen 2011; Carl Zeiss MicroImaging GHBH, Jena,Germany).
4
Quantitative Imaging of Cellular Structures
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Cells grown on Lab-Tek four-well glass chamber slides (NUNC), were incubated in medium or medium containing staurosporine (1 μM) for the indicated times. In some experiments, the cells were incubated with Alexa Fluor 594-conjugated wheat germ agglutinin (2.5 μg/ml) for 10 min at 37 °C and washed twice with HBSS. The cells were fixed with 4% paraformaldehyde and permeabilized with 0.2% Triton X100 PBS or fixed with methanol. The fixed cells were stained with the appropriate Ab and mounted with DAPI-containing mounting medium (Vector Laboratories Ltd, Peterborough, UK). Images were collected using a laser scanning confocal microscope LSM710 (Carl Zeiss, Oberkochen, Germany) equipped with argon (488 nm) and krypton (568 nm) lasers and using a x40 water immersion objective. Images were processed with ZEN 2009 light edition (Carl Zeiss).
5
Quantitative Imaging of Cellular Structures
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Cells grown on Lab-Tek four-well glass chamber slides (NUNC), were incubated in medium or medium containing staurosporine (1 μM) for the indicated times. In some experiments, the cells were incubated with Alexa Fluor 594-conjugated wheat germ agglutinin (2.5 μg/ml) for 10 min at 37 °C and washed twice with HBSS. The cells were fixed with 4% paraformaldehyde and permeabilized with 0.2% Triton X100 PBS or fixed with methanol. The fixed cells were stained with the appropriate Ab and mounted with DAPI-containing mounting medium (Vector Laboratories Ltd, Peterborough, UK). Images were collected using a laser scanning confocal microscope LSM710 (Carl Zeiss, Oberkochen, Germany) equipped with argon (488 nm) and krypton (568 nm) lasers and using a x40 water immersion objective. Images were processed with ZEN 2009 light edition (Carl Zeiss).
6
Immunostaining of S. aureus-challenged cells
7
Fluorescent Imaging of TLR Activation in 661W Cells
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661W cells were cultured on four well glass chamber slides (Fisher Scientific, Rochester, NY) and stimulated with S. aureus and different TLR agonists for 8h. The cells were washed three times with PBS and fixed for 15 min. in 4% paraformaldehyde in PBS. After washing, the cells were permeabilized for 10 min. with an ethanol:acetic acid mixture (2:1) at -20°C and washed once more. The fixed cells were then blocked in 1% (w/v) BSA for 1h at room temperature, followed by incubation with primary antibodies (1:100 dilution) overnight at 4°C. Following removal of the primary antibodies, the cells were washed extensively with PBS and incubated for 1h with specific fluorescein isothiocyanate (FITC)-conjugated secondary antibodies (1:200 dilution) at room temperature. Finally, the cells were again extensively washed with PBS and the slides were mounted in Vectashield anti-fade mounting medium (Vector Laboratories) and visualized using an Eclipse 90i fluorescence microscope (Nikon, Melville, NY).
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