Procedure was previously described (Ramos et al. 2019 (link)). Hela cells were plated at 2.5 × 105 cells on a six-well plate. Cells were transfected 1 d after plating with a 1.25–5 µg of DNA using Lipofectamine 3000. Cells were imaged 24 h post-transfection on an EVOS fluorescence microscopy imaging system (ThermoFisher) quantification. For DNA staining, cells were washed twice with PBS and then incubated for 30 min at 37°C with PBS containing 10% FBS and 1 µM of Hoechst and then imaged. For localization quantification, five images were taken of each well and the number of GFP-positive cells were counted in each of the five frames.
Evos fluorescence microscopy imaging system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The EVOS fluorescence microscopy imaging system is a versatile instrument designed for high-quality fluorescence imaging. It features LED illumination, advanced optics, and a user-friendly interface to capture and analyze fluorescent samples.
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Lab products found in correlation
3 protocols using evos fluorescence microscopy imaging system
1
Hela Cell Transfection and Imaging
2
ROS Quantification in SH-SY5Y Cells
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SH-SY5Y cells were planted into 12-well plates (10 × 104 cells/well) and cultured for 24 h; each group had 3 duplicate wells. The detection was performed strictly in accordance with the manufacturer’s instructions. Briefly, the harvested cells were incubated with diluted DCFH-DA (10 μmol/L final concentration) for 50 min in the dark at 37 °C. After washing with PBS, the images were observed and collected under a fluorescence microscope. Fluorescence was visualized with the EVOS fluorescence microscopy imaging system (Thermo Fisher; Waltham, MA, USA). The ROS levels were expressed as the folds of control.
3
ROS Detection in BV2 Cells
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BV2 cells were treated with 10 μmol/L 2′,7′-dichlorodihydrofluorescein dictate (DCFH-DA, Beyotime, China) for 1 h at 37 °C in the dark. The detection was performed in strict accordance with the manufacturer’s instructions and previous studies [39 (link), 40 (link)]. The fluorescence was visualized by using the EVOS fluorescence microscopy imaging system (Thermo, USA).
Flow cytometry was used to detect ROS production. Firstly, BV2 cells were collected into 15 mL centrifuge tubes and incubated with 10 μmol/L DCFH-DA at 37 °C in the dark for 1 h (mixing cells every 10 min). Subsequently, cells were re-suspended in pre-cooled PBS, washed in pre-cooled PBS three times, and then transferred to 1.5 mL EP tubes for detection by flow cytometry.
Flow cytometry was used to detect ROS production. Firstly, BV2 cells were collected into 15 mL centrifuge tubes and incubated with 10 μmol/L DCFH-DA at 37 °C in the dark for 1 h (mixing cells every 10 min). Subsequently, cells were re-suspended in pre-cooled PBS, washed in pre-cooled PBS three times, and then transferred to 1.5 mL EP tubes for detection by flow cytometry.
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