An adaptive approach for uniform scanning in multifocal multiphoton microscopy with a spatial light modulator

作者：Matsumoto Naoya^*; Okazaki Shigetoshi; Fukushi Yasuko; Takamoto Hisayoshi; Inoue Takashi; Terakawa Susumu

来源：Optics Express, 2014, 22(1): 633-645.

DOI：10.1364/OE.22.000633

We propose high-quality generation of uniform multiple fluorescence spots (MFS) with a spatial light modulator (SLM) and demonstrate uniform laser scanning in multifocal multiphoton microscopy (MMM). The MFS excitation method iteratively updates a computer-generated hologram (CGH) using correction coefficients to improve the fluorescence intensity distribution in a dye solution whose consistency is uniform. This simple correction method can be applied for calibration of the MMM before observation of living tissue. We experimentally demonstrate an improvement of the uniformity of a 10x10 grid of MFS by using a dye solution. After the calibration, we performed laser scanning with two-photon excitation to observe fluorescent polystyrene beads, as well as the gastric gland of a guinea pig specimen.

• 出版日期2014-1-13