School of Applied and Engineering Physics, Cornell University, Ithaca, NY, USA.
CNC Program, Stanford University, Stanford, CA, USA.
Nat Methods. 2018 Oct;15(10):789-792. doi: 10.1038/s41592-018-0115-y. Epub 2018 Sep 10.
Optical imaging through the intact mouse skull is challenging because of skull-induced aberrations and scattering. We found that three-photon excitation provided improved optical sectioning compared with that obtained with two-photon excitation, even when we used the same excitation wavelength and imaging system. Here we demonstrate three-photon imaging of vasculature through the adult mouse skull at >500-μm depth, as well as GCaMP6s calcium imaging over weeks in cortical layers 2/3 and 4 in awake mice, with 8.5 frames per second and a field of view spanning hundreds of micrometers.
通过完整的小鼠颅骨进行光学成像是具有挑战性的，因为颅骨会引起像差和散射。我们发现，与双光子激发相比，三光子激发提供了更好的光学切片，即使我们使用相同的激发波长和成像系统。在这里，我们展示了通过成年小鼠颅骨进行的三光子血管成像，深度超过 500μm，以及在清醒小鼠的皮层 2/3 和 4 层中进行的 GCaMP6s 钙成像，帧率为每秒 8.5 帧，视场跨越数百微米。
