Abstract
Objective: this study proposes and evaluates a technique for in vivo deep-tissue superresolution imaging in the light-scattering mouse brain at up to a 3.5 Hz 2-D imaging rate with a 21×21 μm2 field of view. Methods: we combine the deep-tissue penetration and high imaging speed of resonant laser scanning two-photon (2P) microscopy with the superresolution ability of patterned excitation microscopy. Using high-frequency intensity modulation of the scanned two-photon excitation beam, we generate patterned illumination at the imaging plane. Using the principles of structured illumination, the high-frequency components in the collected images are then used to reconstruct images with an approximate twofold increase in optical resolution. Results: using our technique, resonant 2P superresolution patterned excitation reconstruction microscopy, we demonstrate our ability to investigate nanoscopic neuronal architecture in the cerebral cortex of the mouse brain at a depth of 120 μm in vivo and 210 μm ex vivo with a resolution of 119 nm. This technique optimizes the combination of speed and depth for improved in vivo imaging in the rodent neocortex. Conclusion: this study demonstrates a potentially useful technique for superresolution in vivo investigations in the rodent brain in deep tissue, creating a platform for investigating nanoscopic neuronal dynamics. Significance: this technique optimizes the combination of speed and depth for improved superresolution in vivo imaging in the rodent neocortex.
Original language | English (US) |
---|---|
Pages (from-to) | 232-238 |
Number of pages | 7 |
Journal | IEEE Transactions on Biomedical Engineering |
Volume | 65 |
Issue number | 1 |
DOIs | |
State | Published - Jan 2018 |
Funding
This work was supported in part by the National Institutes of Health under Grants R01EY026078, R24EY022883, and DP3DK108248, in part by the National Science Foundation under Grants DBI-1353952 and CBET-1055379, and in part by the Chicago Biomedical Consortium Catalyst Award. The work of Y. Kozorovitskiy was supported by the William and Bernice E. Bumpus Foundation Innovation Award, the Beckman Young Investigator Award, and the NARSAD Young Investigator Grant and P&S Fund.
Keywords
- Neuron
- Structured illumination
- Super-resolution imaging
- Two-photon microscopy
ASJC Scopus subject areas
- Biomedical Engineering