Institut de Noves Tecnologies de la Imatge (INIT), Universitat Jaume I, Castelló 12080, Spain.
Opt Lett. 2013 Feb 15;38(4):440-2. doi: 10.1364/OL.38.000440.
We demonstrate efficient generation of wide-field fluorescence signals in two-photon microscopy exploiting diffractive optical elements and short pulses by using a dispersion-compensated beam delivery optics module. Computer-generated holograms are codified onto a phase-only spatial light modulator, which allows for arbitrary single-shot patterning of the sample. Spatiotemporal shaping of the pulse is mandatory to overcome spatial chirp and pulse-front tilt effects that spread both in space and time the irradiance patterns, thus limiting not only the spatial resolution but also the signal-to-noise ratio in two-photon microscopy. By using a multipass amplifier delivering 30 fs, 0.8 mJ pulses at 1 kHz repetition rate, we experimentally demonstrated arbitrary single-shot fluorescence irradiance patterns in Rhodamine B.
我们展示了一种通过使用色散补偿光束传输光学模块，利用衍射光学元件和短脉冲在双光子显微镜中高效产生宽场荧光信号的方法。计算机生成的全息图被编码到一个纯相位空间光调制器上，这使得对样品进行任意单次图案化成为可能。脉冲的时空整形是必不可少的，以克服空间啁啾和脉冲前沿倾斜效应，这些效应会在空间和时间上扩展辐照度图案，从而不仅限制了双光子显微镜的空间分辨率，而且还限制了信噪比。通过使用一个多通放大器，我们以 1 kHz 的重复率提供 30 fs、0.8 mJ 的脉冲，在 Rhodamine B 中实验性地演示了任意单次荧光辐照度图案。
