HCT116, HepG2 and SKOV3 cells were seeded into 12-well plates with glass slides, and incubated overnight. Polyplex prepared with Cy5-labled siRNA, Cy5-labled siRNA transfected with Lipo 2000 (3 μL per well) and naked Cy5-labled siRNA with a siRNA concentration of 50 nM were added to each well and incubated for 12 h. After washing with PBS three times, cells were treated with 4% paraformaldehyde for 30 min and stained with DAPI for 10 min, then the distribution of siRNA inside the cells was observed by the Deltavision Elite Microscopy Imaging Systems (GE Healthcare) to evaluate the transfection efficacy.
Deltavision elite microscopy imaging systems
Manufactured by GE Healthcare
Sourced in United States
The Deltavision Elite Microscopy Imaging Systems is a high-performance microscopy imaging platform designed for advanced fluorescence and live-cell imaging applications. It offers a combination of optical and software capabilities to enable detailed analysis of cellular structures and dynamics.
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2 protocols using deltavision elite microscopy imaging systems
1
Visualizing siRNA Transfection Efficiency
2
Cellular Internalization of FITC-Labeled Compounds
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FITC-labeled gelonin and TpG (FITC-Gelonin and FITC-TpG) were used to facilitate cellular internalization study. Three different cancer cell lines ─ human ovarian cancer cell line (SKOV3), human lung adenocarcinoma cell line (A549) and human colon cancer cell line (HCT-8) were seeded on glass coverslips in a 12-well plate and incubated for 24 h. Next, the medium was replaced with fresh medium (pH 7.4 or 6.5) containing FITC-Gelonin or FITC-TpG (with a concentration of 1 μM) and incubated for 12 h. The cells were washed three times with PBS, stained with DAPI for cell nuclei, and observed by Deltavision Elite Microscopy Imaging Systems (GE Healthcare).
For flow cytometric assay, SKOV3, A549 and HCT-8 cells were seeded into 12-well plates and incubated for 24 h, FITC-Gelonin or FITC-TpG (1 μM) in fresh medium (pH 7.4 or 6.5) was added and incubated for 12 h. The cells were harvested by trypsinization, washed twice with cold PBS, filtered through nylon mesh, and subjected to flow cytometric analysis using a Beckman Coulter CytoFLEX flow cytometer (Beckman Coulter, USA).
For flow cytometric assay, SKOV3, A549 and HCT-8 cells were seeded into 12-well plates and incubated for 24 h, FITC-Gelonin or FITC-TpG (1 μM) in fresh medium (pH 7.4 or 6.5) was added and incubated for 12 h. The cells were harvested by trypsinization, washed twice with cold PBS, filtered through nylon mesh, and subjected to flow cytometric analysis using a Beckman Coulter CytoFLEX flow cytometer (Beckman Coulter, USA).
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