Samples of 500,000 Kc167 cells were harvested, spun down, and resuspended in ice-cold phosphate-buffered saline (PBS) with 0.1% Triton X-100 (PBST) and 0.1% formaldehyde and incubated for 10 min on ice. Cells were spun down and resuspended in ice-cold PBST. Cells were then spun down onto a clean glass slide (Fisherbrand Superfrost Plus Microscope Slides; ThermoFisher Scientific, catalog no. 12-550-15) using a Thermo Scientific Shandon Cytospin 4. Slides were washed twice with 50 ml of 1× PBS each time and placed in a humid chamber with 1 ml of blocking solution (2.5% bovine serum albumin in 1× PBST) for 30 min of pre-blocking. The blocking buffer was then drained, and primary antibodies were added for incubation overnight at 4°C. Slides were then washed twice with 50 ml of 1× PBS and incubated with secondary antibodies for 2 hours at RT. Slides were then washed twice with 50 ml of 1× PBS and mounted with ProLong Diamond Mounting Media (Thermo Fisher Scientific, P36965). Cells were imaged on an EVOS auto 2.0 microscope (Thermo Fisher Scientific).
Evos auto 2.0 microscope
Manufactured by Thermo Fisher Scientific
The EVOS auto 2.0 is a digital microscope designed for high-quality cell imaging. It features a motorized stage and automated focus control for precise sample positioning and capture. The system is capable of bright-field, phase-contrast, and fluorescence imaging.
Automatically generated - may contain errors
2 protocols using evos auto 2.0 microscope
1
Immunofluorescence Staining of Kc167 Cells
2
Immunofluorescence Staining Kc167 Cells
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Samples of 500,000 Kc167 cells were harvested, spun down and resuspended in ice cold PBS with 0.1% Triton and 0.1% formaldehyde and incubated for 10 minutes on ice. Cells were spun down and resuspended in ice cold PBS with 0.1% Triton-X100. Cells were then spun down onto a clean glass slide (Fisherbrand Superfrost Plus Microscope Slides; ThermoFisher Scientific, cat. no. 12–550-15) using a ThermoScientific Shandon Cytospin 4. Slides were washed twice with 50 mL 1× PBS each time and placed in a humid chamber with 1 mL of blocking solution (2.5% BSA in 1× PBST) for 30 minutes of pre-blocking. Blocking buffer was then drained, and primary antibodies were added for incubation overnight at 4 °C. Slides were then washed twice with 50 mL 1× PBS and incubated with secondary antibodies for 2 hours at RT. Slides were then washed twice with 50 mL 1× PBS and mounted with ProLong Diamond Mounting Media (Thermo Fisher Scientific: P36965). Cells were imaged on an EVOS auto 2.0 microscope (ThermoFisher).
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