Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution

Meddens, Marjolein B.  M.; Liu, Sheng; Finnegan, Patrick S.; Edwards, Thayne L.; James, Conrad D.; Lidke, Keith A.

doi:10.1364/BOE.7.002219

Title: Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution

Journal Article · Fri Jan 01 00:00:00 EST 2016 · Biomedical Optics Express

DOI:https://doi.org/10.1364/BOE.7.002219· OSTI ID:1259471

Meddens, Marjolein B. M. ^[1]; Liu, Sheng ^[2]; Finnegan, Patrick S. ^[3]; Edwards, Thayne L. ^[3]; James, Conrad D. ^[3]; Lidke, Keith A. ^[1]

Univ. of New Mexico, Albuquerque, NM (United States)
Univ. of New Mexico, Albuquerque, NM (United States); Purdue Univ., West Lafayette, IN (United States)
Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

Here, we have developed a method for performing light-sheet microscopy with a single high numerical aperture lens by integrating reflective side walls into a microfluidic chip. These 45° side walls generate light-sheet illumination by reflecting a vertical light-sheet into the focal plane of the objective. Light-sheet illumination of cells loaded in the channels increases image quality in diffraction limited imaging via reduction of out-of-focus background light. Single molecule super-resolution is also improved by the decreased background resulting in better localization precision and decreased photo-bleaching, leading to more accepted localizations overall and higher quality images. Moreover, 2D and 3D single molecule super-resolution data can be acquired faster by taking advantage of the increased illumination intensities as compared to wide field, in the focused light-sheet.

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Research Organization:: Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

Sponsoring Organization:: USDOE National Nuclear Security Administration (NNSA)

Grant/Contract Number:: AC04-94AL85000

OSTI ID:: 1259471

Report Number(s):: SAND2016-4996J; 640849

Journal Information:: Biomedical Optics Express, Vol. 7, Issue 6; ISSN 2156-7085

Publisher:: Optical Society of AmericaCopyright Statement

Country of Publication:: United States

Language:: English

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