The cells were cultured in glass coverslip-bottomed culture dishes (MatTek, Ashland, MA) to 50–80% confluence. After the culture medium was aspirated, the cells were rinsed with PBS twice, fixed with 3.7% paraformaldehyde at 25 °C for 10 min, and permeabilized with 0.2% Triton X-100 in PBS at 25 °C for 10 min. After washing with PBS, the cells were incubated with primary antibody at 8 °C overnight. The cells were washed with PBS three times and incubated with a fluorescent dye-conjugated secondary antibody and phalloidin with at 37 °C for 1–2 h. After washed with PBS three times, fluorescent staining of the cells were visualized under Zeiss LSM710 confocal microscope or Nikon inverted fluorescent microscope.
Glass coverslip bottomed culture dishes
Manufactured by MatTek
Glass coverslip-bottomed culture dishes are a type of cell culture equipment used for microscopic observation and analysis. These dishes feature a thin glass coverslip as the bottom, allowing for high-quality imaging and visualization of cells under a microscope.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using glass coverslip bottomed culture dishes
1
Immunofluorescence Cell Imaging Protocol
2
Immunofluorescence Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were cultured in glass coverslip-bottomed culture dishes (MatTek, Ashland, MA) to 50-80% confluence. After the culture medium was aspirated, the cells were rinsed with PBS twice, fixed with 3.7% paraformaldehyde at 25°C for 30 min, and permeabilized with 0.2% Triton X-100 in PBS at 25°C for 20 min. After washing with TBST, the cells were incubated with primary antibody at 37°C for 1 h. Then the cells were washed with TBST three times and incubated with secondary antibody that was conjugated with a fluorescent dye at 37°C for 1 h. Finally, the cells were washed with TBST three times, and the immunofluorescence staining was visualized under a Nikon inverted fluorescent microscope. The nuclei were stained with DAPI.
3
Immunofluorescence Microscopy Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For IF microscopy, the cells were cultured in glass coverslip-bottomed culture dishes (MatTek, Ashland, MA). After the culture medium was aspirated, the cells were rinsed with PBS twice, fixed in 4% paraformaldehyde, and permeabilized with 0.5% Triton X-100 for 15 minutes. The slides were then incubated with a primary antibody in blocking solution overnight at 4°C in a humidified chamber. The glass slides were then washed three times in PBS and incubated with Alexa Fluor 568- or Alexa Fluor 488 -conjugated second antibody for 1 h at room temperature in a humidified chamber. Finally, the cover slips were incubated with 40, 60-diamidino-2-phenylindole (Sigma-Aldrich) for 15 min, and images were obtained with a phase-contrast and confocal microscopy.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!