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Direct wavefront sensing for high-resolution in vivo imaging in scattering tissue.

Wang K, Sun W, Richie CT, Harvey BK, Betzig E, Ji N - Nat Commun (2015)

Bottom Line: Adaptive optics by direct imaging of the wavefront distortions of a laser-induced guide star has long been used in astronomy, and more recently in microscopy to compensate for aberrations in transparent specimens.Here we extend this approach to tissues that strongly scatter visible light by exploiting the reduced scattering of near-infrared guide stars.The method enables in vivo two-photon morphological and functional imaging down to 700 μm inside the mouse brain.

View Article: PubMed Central - PubMed

Affiliation: Janelia Research Campus, Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, Virginia 20147, USA.

ABSTRACT
Adaptive optics by direct imaging of the wavefront distortions of a laser-induced guide star has long been used in astronomy, and more recently in microscopy to compensate for aberrations in transparent specimens. Here we extend this approach to tissues that strongly scatter visible light by exploiting the reduced scattering of near-infrared guide stars. The method enables in vivo two-photon morphological and functional imaging down to 700 μm inside the mouse brain.

No MeSH data available.


Related in: MedlinePlus

Exemplary spatial variation of sample-induced aberrations in the cortex of a living mouse.(a) SH images and (b) their corresponding corrective wavefronts obtained at pia surface and three different regions (i, ii and iii) 400 μm below pia of a Thy1-YFPH mouse. (c) Images of regions i, ii and iii obtained with the corrective wavefronts in b, respectively. Scale bar, 20 μm. The inset (‘map of imaging locations') shows the relative positions of regions i, ii and iii. Scale bar, 100 μm (inset).
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f2: Exemplary spatial variation of sample-induced aberrations in the cortex of a living mouse.(a) SH images and (b) their corresponding corrective wavefronts obtained at pia surface and three different regions (i, ii and iii) 400 μm below pia of a Thy1-YFPH mouse. (c) Images of regions i, ii and iii obtained with the corrective wavefronts in b, respectively. Scale bar, 20 μm. The inset (‘map of imaging locations') shows the relative positions of regions i, ii and iii. Scale bar, 100 μm (inset).

Mentions: Even though a single correction can improve image quality over a field of hundreds of microns in the mouse cortex512, the best performance is still obtained when the aberration is measured at the imaging location. To demonstrate, we measured aberrations at the surface of the mouse cortex as well as three locations at 400-μm depth that were laterally separated by 200–400 μm (Fig. 2). Although the three wavefronts measured at 400-μm depth were similar, in all three cases the highest signal and greatest contrast recovery occurred when the locally measured correction was employed. Therefore, to have diffraction-limited images over a large field of view (for example, 1 × 1 mm2), frequent AO corrections are likely needed. Our method is well suited for this eventuality, thanks to its high correction speed (milliseconds to seconds; Supplementary Table 1).


Direct wavefront sensing for high-resolution in vivo imaging in scattering tissue.

Wang K, Sun W, Richie CT, Harvey BK, Betzig E, Ji N - Nat Commun (2015)

Exemplary spatial variation of sample-induced aberrations in the cortex of a living mouse.(a) SH images and (b) their corresponding corrective wavefronts obtained at pia surface and three different regions (i, ii and iii) 400 μm below pia of a Thy1-YFPH mouse. (c) Images of regions i, ii and iii obtained with the corrective wavefronts in b, respectively. Scale bar, 20 μm. The inset (‘map of imaging locations') shows the relative positions of regions i, ii and iii. Scale bar, 100 μm (inset).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4490402&req=5

f2: Exemplary spatial variation of sample-induced aberrations in the cortex of a living mouse.(a) SH images and (b) their corresponding corrective wavefronts obtained at pia surface and three different regions (i, ii and iii) 400 μm below pia of a Thy1-YFPH mouse. (c) Images of regions i, ii and iii obtained with the corrective wavefronts in b, respectively. Scale bar, 20 μm. The inset (‘map of imaging locations') shows the relative positions of regions i, ii and iii. Scale bar, 100 μm (inset).
Mentions: Even though a single correction can improve image quality over a field of hundreds of microns in the mouse cortex512, the best performance is still obtained when the aberration is measured at the imaging location. To demonstrate, we measured aberrations at the surface of the mouse cortex as well as three locations at 400-μm depth that were laterally separated by 200–400 μm (Fig. 2). Although the three wavefronts measured at 400-μm depth were similar, in all three cases the highest signal and greatest contrast recovery occurred when the locally measured correction was employed. Therefore, to have diffraction-limited images over a large field of view (for example, 1 × 1 mm2), frequent AO corrections are likely needed. Our method is well suited for this eventuality, thanks to its high correction speed (milliseconds to seconds; Supplementary Table 1).

Bottom Line: Adaptive optics by direct imaging of the wavefront distortions of a laser-induced guide star has long been used in astronomy, and more recently in microscopy to compensate for aberrations in transparent specimens.Here we extend this approach to tissues that strongly scatter visible light by exploiting the reduced scattering of near-infrared guide stars.The method enables in vivo two-photon morphological and functional imaging down to 700 μm inside the mouse brain.

View Article: PubMed Central - PubMed

Affiliation: Janelia Research Campus, Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, Virginia 20147, USA.

ABSTRACT
Adaptive optics by direct imaging of the wavefront distortions of a laser-induced guide star has long been used in astronomy, and more recently in microscopy to compensate for aberrations in transparent specimens. Here we extend this approach to tissues that strongly scatter visible light by exploiting the reduced scattering of near-infrared guide stars. The method enables in vivo two-photon morphological and functional imaging down to 700 μm inside the mouse brain.

No MeSH data available.


Related in: MedlinePlus