Highly inclined plane illumination microscopy with extended imaging depth (Conference Presentation)

Jinhan Ren; Kyu Young Han

doi:10.1117/12.2544314

9 March 2020 Highly inclined plane illumination microscopy with extended imaging depth (Conference Presentation)

Jinhan Ren, Kyu Young Han

Proceedings Volume 11246, Single Molecule Spectroscopy and Superresolution Imaging XIII; 112460J (2020) https://doi.org/10.1117/12.2544314
Event: SPIE BiOS, 2020, San Francisco, California, United States

Abstract

Confocal microscopy features a good sectioning capability, which makes it essential for high-resolution 3D biological imaging. However, a tightly focused excitation beam inevitably leads to irreversible photodamage to live specimens, such as photobleaching and phototoxicity, and point-by-point scanning mechanism hampers its applications in fast volumetric imaging. As an alternative approach, selective plane illumination microscopy has shown outstanding performance in long-term imaging of embryonic development and neuronal activities attributed to its capabilities of intrinsic sectioning, gentle excitation and fast imaging. One drawback of SPIM is that the arrangement of two closely placed objectives greatly restricts the geometry of sample holders and the available numerical aperture (NA) for effective fluorescence collection. Here, we propose a highly-inclined plane illumination with a single high NA objective using for both excitation and detection. Unlike the requirement of two relay objective units in remote focusing imaging, an adaptive optical device serving as a flexible wavefront modulator compensates the systematical aberrations induced by optical components and effectively extended the imaging depth by generating an elongated point spread function (PSF). Our technique is applicable to single-molecule tracking and super-resolution imaging in live cells. Moreover, the adaptive optics can be replaced with a transmitted phase mask to enhance the effective fluorescent collection and simplify the system alignment effort.

Conference Presentation

Citation Download Citation

Jinhan Ren and Kyu Young Han "Highly inclined plane illumination microscopy with extended imaging depth (Conference Presentation)", Proc. SPIE 11246, Single Molecule Spectroscopy and Superresolution Imaging XIII, 112460J (9 March 2020); https://doi.org/10.1117/12.2544314

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KEYWORDS

Microscopy

Objectives

Imaging systems

Optical components

Point spread functions

3D image processing

Confocal microscopy

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