Zeiss Axiovert 200 system was used to observe cell shape changes with optostimulation. The T-Cube LED Driver (Thorlabs, Newton, NJ, LEDD1B) and Blue (470 nm) Fiber-Coupled High-Power LED (Thorlabs, Newton, NJ, M470F1) were used to generate optostimulation (~24 mW/mm2) with optical fiber (Thorlabs, Newton, NJ, FT400EMT). Cell images were recorded continuously with a 20 s interval. Additional cell contraction was achieved with a 5 min exposure to 20 mM KCl. Data were accepted for further analysis if the features of VSM cells were seen with YFP signal and KCl response. Cell shortening was measured along its longitudinal axis by the ImageJ 1.48 software.
Axiovert 200 system
Manufactured by Zeiss
The Axiovert 200 system is a inverted microscope platform from Zeiss. It is designed for applications that require a stable and versatile imaging setup. The system provides a range of optical configurations and accessories to support various microscopy techniques.
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Lab products found in correlation
2 protocols using axiovert 200 system
1
Optostimulation Cell Shape Analysis
2
Cell Labeling and Imaging Protocol
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BT-474 cells at the third or fourth passage (5 × 105 cells/well in 0.5 mL of 46X medium) were placed in four-well chamber slides and grown for 24–48 h to 70–80 confluency. Cells were first incubated with Tz(TCO)6(AF-488)4 (20 μg/mL, 130 nM) in PBS+ (PBS supplemented with 0.5% BSA) at room temperature for 20 min. Pre-labeled cells were treated with reactive Alb(Peg4-Tt)15(Rhod)4 or non-reactive Alb(Rhod)4 and incubated at room temperature for 15 min. Treated cells were then incubated in fresh 46-X medium for 3 h at room temperature, or 37 °C, and fixed by 4% PFA in PBS. Cells were counterstained by Hoechst 33342 (1 μg/mL in dH2O) and wet-mounted for confocal microscopic imaging on a Zeiss Axiovert 200 system equipped with an LSM 510-Meta confocal module.
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