For immunofluorescence staining, keratinocytes on PDMS substrates were fixed in 4% paraformaldehyde (Electron Microscopy Science) and permeabilized in 0.5% Triton X-100 (Sigma-Aldrich). For actin and nucleus staining, the cells were incubated with Alexa-488-labeled phalloidin (Invitrogen) and Hoechst 33342 (Sigma-Aldrich) in 1% BSA (Sigma-Aldrich) blocking solution. For focal adhesion staining, the cells were incubated with a focal adhesion kinase (Proteintech, Rosemont, IL, USA) primary antibody for 1 h at room temperature. After washing, the cells were incubated for 1 h with Alexa-568-labeled goat anti-rabbit secondary antibody (Thermo Fisher Scientific). All images were obtained using an upright microscope (Nikon) with Plan Fluor 20× (NA 0.5) and S Plan Fluor 40× (NA 0.6) objectives.
S plan fluor 40
Manufactured by Nikon
Sourced in Japan
The S Plan Fluor 40× is an objective lens designed for microscopy applications. It provides a magnification of 40× and is suitable for use with a variety of sample types. The lens features a plan-apochromatic optical design, which ensures high-quality, distortion-free images across the entire field of view.
Automatically generated - may contain errors
Lab products found in correlation
Upright microscope, by Nikon (2 mentions)
Plan fluor 20, by Nikon (2 mentions)
Alexa 488 labeled phalloidin, by Thermo Fisher Scientific (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Hoechst 33342, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions)
Calcein am, by Thermo Fisher Scientific (1 mentions)
2 protocols using s plan fluor 40
1
Immunofluorescence Staining of Keratinocytes
2
Quantifying Cell Morphology and Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were fixed in 4% paraformaldehyde and permeabilized in 0.5%
Triton X-100 (Sigma-Aldrich, USA). For actin and nucleus staining,
the cells were incubated with Alexa-488-labeled phalloidin (Invitrogen,
USA) and Hoechst 33342 (Sigma-Aldrich, USA) in PBS with 1% bovine
serum albumin (Sigma-Aldrich, USA). All images were obtained using
an upright microscope (Nikon, Japan) with Plan Fluor 20× (NA
0.5) and S Plan Fluor 40× (NA 0.6) objectives. Quantification
analysis was performed to measure the cell area, perimeter, and aspect
ratio using the open source software Cell Profiler.52 (link) For a live–dead viability test, an ethidium homodimer
in combination with Calcein-AM (Invitrogen, USA) was used.
Triton X-100 (Sigma-Aldrich, USA). For actin and nucleus staining,
the cells were incubated with Alexa-488-labeled phalloidin (Invitrogen,
USA) and Hoechst 33342 (Sigma-Aldrich, USA) in PBS with 1% bovine
serum albumin (Sigma-Aldrich, USA). All images were obtained using
an upright microscope (Nikon, Japan) with Plan Fluor 20× (NA
0.5) and S Plan Fluor 40× (NA 0.6) objectives. Quantification
analysis was performed to measure the cell area, perimeter, and aspect
ratio using the open source software Cell Profiler.52 (link) For a live–dead viability test, an ethidium homodimer
in combination with Calcein-AM (Invitrogen, USA) was used.
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