The unfractionated endometrial stromal cells or eMSCs (8000–10,000 cells) were resuspended in growth medium and transferred to slides coated with 3-aminopropyl-triethoxysilane using a Shadon Cytospin Centrifuge (Thermo Electron, Waltham, USA) with centrifugation at 7500 rpm for 10 min followed by fixation in 4% paraformaldehyde for 20 min. Permeabilization was performed using 0.1% Triton-X 100 for 10 min and blocked with the corresponding serum for 30 min at room temperature. The slides were then incubated with the primary antibody (Additional file: Table S4) overnight at 4 °C, incubated with the secondary Alexa fluor donkey anti-rabbit 568 antibody (Thermo Scientific) for 1 h at room temperature. The cell nuclei were detected by DAPI (Thermo Scientific). Images were captured with a LSM 700 inverted confocal microscope and a LSM ZEN 2010 software (Carl Zeiss, Munich, Germany) at the Centre for PanorOmic Sciences (CPOS) imaging and Flow Cytometry Core, The University of Hong Kong.
Shadon cytospin centrifuge
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Shadon Cytospin Centrifuge is a laboratory instrument designed for the preparation of cytological samples. Its core function is to concentrate cells or other particulate matter onto a microscope slide, enabling their examination and analysis.
Automatically generated - may contain errors
3 protocols using shadon cytospin centrifuge
1
Immunofluorescent Staining of eMSCs
2
Immunostaining of Endometrial Stromal Cells
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The unfractionated endometrial stromal cells or eMSCs (8000 -10,000 cells) were resuspended in growth medium and transferred to slides coated with 3-aminopropyl-triethoxy silane using a Shadon Cytospin Centrifuge (Thermo Electron, Waltham, USA) with centrifugation at 7500 rpm for 10 minutes followed by xation in 4% paraformaldehyde for 20 minutes. Permeabilization was performed using 0.1% Triton-X 100 for 10 minutes and blocked with the corresponding serum for 30 minutes at room temperature. The slides were then incubated with the primary antibody (supplementary data Table S4) overnight at 4°C, incubated with the secondary Alexa uor donkey anti-rabbit 568 antibody (Thermo Scienti c) for 1 hour at room temperature. The cell nuclei were detected by DAPI (Thermo Scienti c). Images were captured with a LSM 700 inverted confocal microscope and a LSM ZEN 2010 software (Carl Zeiss, Munich, Germany) at the Centre for PanorOmic Sciences (CPOS) imaging and Flow Cytometry Core, The University of Hong Kong.
3
Immunofluorescent Staining of eMSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The unfractionated endometrial stromal cells or eMSCs (8000 -10,000 cells) were resuspended in growth medium and transferred to slides coated with 3-aminopropyl-triethoxy silane using a Shadon Cytospin Centrifuge (Thermo Electron, Waltham, USA) with centrifugation at 7500 rpm for 10 minutes followed by xation in 4% paraformaldehyde for 20 minutes. Permeabilization was performed using 0.1% Triton-X 100 for 10 minutes and blocked with the corresponding serum for 30 minutes at room temperature. The slides were then incubated with the primary antibody (Additional le: Table S4) overnight at 4°C, incubated with the secondary Alexa uor donkey anti-rabbit 568 antibody (Thermo Scienti c) for 1 hour at room temperature. The cell nuclei were detected by DAPI (Thermo Scienti c).
Images were captured with a LSM 700 inverted confocal microscope and a LSM ZEN 2010 software (Carl Zeiss, Munich, Germany) at the Centre for PanorOmic Sciences (CPOS) imaging and Flow Cytometry Core, The University of Hong Kong.
Images were captured with a LSM 700 inverted confocal microscope and a LSM ZEN 2010 software (Carl Zeiss, Munich, Germany) at the Centre for PanorOmic Sciences (CPOS) imaging and Flow Cytometry Core, The University of Hong Kong.
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