Fluorescence measurements were obtained using IX71 inverted microscope (Olympus, Tokyo, Japan) attached with a 60× (numerical aperture 0.9) water-immersion objective lens as previously reported14 (link). The signal was detected by a cool CCD video camera Ixon (Andor Co., Tokyo, Japan) coupled to Lambda-10.2 (Sutter Instruments, CA), and excitation was provided by Sutter DG-4 and 175-W Xenon-Arc lamp (Sutter Instruments, Navato, CA). For the experiments, cover slips were placed in an imaging chamber (RC-42LP and RC-43C; Warner Instruments, Hamden, CT) mounted on the stage of the inverted microscope and maintained at 37 °C with TC-344B thermo-warmer for chamber and TA-29 thermo-warmer for perfusate (Warner Instruments, Hamden, CT). The changes in fluorescence intensity were quantified using MetaFluor imaging software (Universal Imaging Co., Ypsilanti, MI) for each experiment. To exclude the possibility of selection bias of the measured MCs, the fluorescence intensity of all living mesangial cells in each cover glass was measured.
175 w xenon arc lamp
Manufactured by Sutter Instruments
Sourced in Japan
The 175-W Xenon-Arc lamp is a high-intensity light source that generates illumination using a xenon gas-filled arc. It is designed to provide a broad spectrum of light output, making it suitable for various applications that require intense, uniform illumination.
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2 protocols using 175 w xenon arc lamp
1
Fluorescence Imaging of Mesangial Cells
2
Retinal Light Stimulation Protocol
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Full-field light stimulation was provided by a 175-W xenon arc lamp (Sutter Instruments). Calibrated neutral density filters and narrowband interference filters were used to control light density and stimulus wavelength, respectively. Retinas were stimulated with full-field unpolarized monochromatic (500 nm, 10 nm half-width) light, and the duration of the stimulus was 500 ms. The intensity of the unattenuated stimulus at 500 nm was 2.18 × 10−4 W/cm2 or 5.49 × 1014 photons/cm2 per second, converted to 2.06 × 106R*/rod/s) [for details, see (43 (link))]. Intervals between light stimuli were adjusted so that the baseline of activity was able to fully recover to its initial level before the next stimulus. For the experiments depicted in Fig. 6 , we used a 500-nm light-emitting diode (LED) as the light source. The LED signal was delivered either at 1 Hz or at 10 Hz (square signal).
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