Adaptive optics can correct for optical aberrations. We developed multi-pupil adaptive optics (MPAO), which enables simultaneous wavefront correction over a field of view of 450 × 450 μm2 and expands the correction area to nine times that of conventional methods. MPAO's ability to perform spatially independent wavefront control further enables 3D nonplanar imaging. We applied MPAO to in vivo structural and functional imaging in the mouse brain.
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This work was funded by NIH grant no. 1U01NS094341-01 and Purdue University. The authors thank W. Gan for valuable discussion and advice; G. Holtom and B. Wei for their help on manuscript preparation; and the Howard Hughes Medical Institute for equipment support. J.-H.P. thanks the NRF (grant no. 2016R1C1B2015130) for support during the manuscript preparation; and L.K. thanks the NSFC (grant no. 61327902) for support during manuscript revision.
Integrated supplementary information
We show the MIP of microglia along z axis (450 × 450 × 40 μm3 volume at the depth of 330-370 μm) recorded by system correction with MPAO, full correction with MPAO, full correction with an averaged wavefront (full average), and full correction with the wavefront measured from one region (full single, nine groups in sequence).
MIP along z axis of the 450 × 450 × 50 μm3 volume at 0-50 μm under the dura. Using MPAO with full correction, we performed large FOV volumetric imaging of resting state microglia. In addition to the motion of microglia and dendritic cells, the video also captured GFP-expressing leukocytes trafficking in brain vasculatures.
MIP along z axis of the 450 × 450 × 35 μm3 volume at the depth of 330-365 μm. Using laser ablation, we introduced tissue damage, and employed MPAO with full correction to record the activated microglia dynamics.
The image volume was 450×450×550 μm3. We show the volume data with system and with full correction for comparison.
In the video, we show the partial data of the calcium transient in Supplementary Fig. 10. The imaging FOV is 450 × 450 μm 2.
The image in Supplementary Fig. 11d is extracted from this video. Magenta: the 3D image stacks. Green: the planes defined by the defocusing wavefront in nonplanar imaging. The imaging FOV is 450 × 450 μm2.
The plot in Fig. 3e is extracted from this video. The imaging FOV is 450 × 450 μm 2.
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Archives of Pharmacal Research (2019)