We avoided measurements within the same cells to minimize photo-bleaching of insulin signal caused by repeated exposure to laser beams.
The level of [Ca2+]c was estimated with YC2.1 at every 10 minute in order to minimize photo-bleaching: with this slow image acquisition rate, the presence or the absence of Ca2+ spikes could not be determined [16].
To minimize photo-bleaching, the intensity of light was reduced to 1 4% by intensity control.
For image acquisition, a short exposure time (200 ms) and low-intensity excitation light were applied to minimize photo-bleaching.
All manipulations involving RA were conducted under subdued lighting or yellow lights in order to minimize photo-oxidation.
Although significantly faster imaging rates are possible, these conditions were chosen to minimize photo-bleaching and phototoxicity, while specifically selecting stably bound (>3 s) molecules.
CARS also enables detection in the presence of one-photon fluorescence, 3-D sectioning, and penetration to a depth of ~0.4 mm while minimizing photo damage [ 19– 219.
Compared to previous works where conventional fluorescence microscopy was used, our approach has been demonstrated to offer all the advantages of multi-photon imaging, providing substantial enhancement in both spatial and temporal resolutions while minimizing photo-damages.
Further, the use of a helium cryostream is compatible with XAS measurements at the sulfur K-edge and has the advantage of minimizing thiol oxidation due to air-exposure, minimizing spectral distortion due to X-ray absorption by argon, and also minimizing photo-oxidation of sulfur (see Supporting Information Figure 1).
43, 62 It is important to advise patients using tacrolimus ointment to avoid a solarium, to minimize exposure to sunlight, and to use sunscreen, even though it is not a photo-sensitizer.
