Photonics Research, Volume. 9, Issue 6, 1144(2021)
High-resolution two-photon transcranial imaging of brain using direct wavefront sensing
Fig. 1. Optimization of the excitation NA and improved wavefront sensing algorithm. (a) Schematic illustration of a high-NA objective in the overfilled (top) and underfilled (bottom) configuration. (b) Monte Carlo simulated SHWS images from the guide star at 250 μm below the thinned skull with full NA and reduced NA of the objective. The dashed circles indicate the apertures for the effective NA of 1.05 and 0.7, respectively. Details are shown in Fig. S2 in Ref. [21]. (c) Simulated wavefront errors and theoretical lateral resolution as functions of effective NA. (d) Representative guide star images of YFP fluorescence in Thy1-YFP mice when the objective was overfilled (left) and underfilled (right) at 250 μm below the thinned skull. Details are shown in Fig. S3 in Ref. [21]. (e) Percentages of bad spots in the Shack–Hartmann spot image for overfilled (blue) and underfilled (red) configurations. A bad spot is the one with poor signal quality (SBR less than 1.2) to make its center unidentifiable. (f) Top: depth-coded images of the NIR-dye labelled microvascular vessels for direct wavefront sensing. Two segments of vessels at different depths (solid line and dashed line labelled with ls 1 and ls 2) were line scanned for wavefront measurement. Scale bar: 10 μm. Bottom: two-photon images of YFP labelled dendrite with system correction only. (g) Top row: guide star images on the SHWS with only ls 1 (left) and our algorithm (right). The right-top corners display the corresponding corrective wavefronts. Bottom row: the corresponding AO corrected images. Details are presented in Fig. S7 in Ref. [21]. Scale bar: 5 μm. (h) Comparisons of the detailed structures of the numbered spots in (g). (i) Intensity profiles along the dashed lines in the bottom panel of (f) and (g).
Fig. 2. AO improves structural and functional brain imaging
Fig. 3. Study of microglial activities in AD mice and neuron–microglial interactions following precise laser micro-lesion. (a)
Fig. 4. AO recovers high-resolution imaging of the cortex through an optical clearing window. (a)
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Congping Chen, Zhongya Qin, Sicong He, Shaojun Liu, Shun-Fat Lau, Wanjie Wu, Dan Zhu, Nancy Y. Ip, Jianan Y. Qu. High-resolution two-photon transcranial imaging of brain using direct wavefront sensing[J]. Photonics Research, 2021, 9(6): 1144
Category: Imaging Systems, Microscopy, and Displays
Received: Jan. 20, 2021
Accepted: Mar. 14, 2021
Published Online: Jul. 7, 2021
The Author Email: Jianan Y. Qu (eequ@ust.hk)