Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution
Abstract
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.
- Authors:
-
- 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)
- Publication Date:
- Research Org.:
- Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
- Sponsoring Org.:
- USDOE National Nuclear Security Administration (NNSA)
- OSTI Identifier:
- 1259471
- Report Number(s):
- SAND2016-4996J
Journal ID: ISSN 2156-7085; 640849
- Grant/Contract Number:
- AC04-94AL85000
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Biomedical Optics Express
- Additional Journal Information:
- Journal Volume: 7; Journal Issue: 6; Journal ID: ISSN 2156-7085
- Publisher:
- Optical Society of America
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 47 OTHER INSTRUMENTATION
Citation Formats
Meddens, Marjolein B. M., Liu, Sheng, Finnegan, Patrick S., Edwards, Thayne L., James, Conrad D., and Lidke, Keith A. Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution. United States: N. p., 2016.
Web. doi:10.1364/BOE.7.002219.
Meddens, Marjolein B. M., Liu, Sheng, Finnegan, Patrick S., Edwards, Thayne L., James, Conrad D., & Lidke, Keith A. Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution. United States. https://doi.org/10.1364/BOE.7.002219
Meddens, Marjolein B. M., Liu, Sheng, Finnegan, Patrick S., Edwards, Thayne L., James, Conrad D., and Lidke, Keith A. Fri .
"Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution". United States. https://doi.org/10.1364/BOE.7.002219. https://www.osti.gov/servlets/purl/1259471.
@article{osti_1259471,
title = {Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution},
author = {Meddens, Marjolein B. M. and Liu, Sheng and Finnegan, Patrick S. and Edwards, Thayne L. and James, Conrad D. and Lidke, Keith A.},
abstractNote = {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.},
doi = {10.1364/BOE.7.002219},
journal = {Biomedical Optics Express},
number = 6,
volume = 7,
place = {United States},
year = {Fri Jan 01 00:00:00 EST 2016},
month = {Fri Jan 01 00:00:00 EST 2016}
}
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