Hyperspectral stimulated emission depletion microscopy and methods of use thereof

Timlin, Jerilyn A; Aaron, Jesse S

Hyperspectral stimulated emission depletion microscopy and methods of use thereof

Patent · 01 April 2014

OSTI ID:1128705

Timlin, Jerilyn A; Aaron, Jesse S

A hyperspectral stimulated emission depletion ("STED") microscope system for high-resolution imaging of samples labeled with multiple fluorophores (e.g., two to ten fluorophores). The hyperspectral STED microscope includes a light source, optical systems configured for generating an excitation light beam and a depletion light beam, optical systems configured for focusing the excitation and depletion light beams on a sample, and systems for collecting and processing data generated by interaction of the excitation and depletion light beams with the sample. Hyperspectral STED data may be analyzed using multivariate curve resolution analysis techniques to deconvolute emission from the multiple fluorophores. The hyperspectral STED microscope described herein can be used for multi-color, subdiffraction imaging of samples (e.g., materials and biological materials) and for analyzing a tissue by Forster Resonance Energy Transfer ("FRET").

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

Sponsoring Organization:: USDOE

DOE Contract Number:: AC04-94AL85000;

Assignee:: Sandia Corporation (Albuquerque, NM)

Patent Number(s):: 8,686,363

Application Number:: 13/609,057

OSTI ID:: 1128705

Country of Publication:: United States

Language:: English

References (7)

Weighting hyperspectral image data for improved multivariate curve resolution results Jones, Howland D. T.; Haaland, David M.; Sinclair, Michael B. Journal of Chemometrics, Vol. 22, Issue 9, p. 482-490 https://doi.org/10.1002/cem.1170	journal	September 2008
Preprocessing strategies to improve MCR analyses of hyperspectral images Jones, Howland D.T.; Haaland, David M.; Sinclair, Michael B. Chemometrics and Intelligent Laboratory Systems, Vol. 117, p. 149-158 https://doi.org/10.1016/j.chemolab.2012.01.011	journal	August 2012
Förster resonance energy transfer – A spectroscopic nanoruler: Principle and applications Sahoo, Harekrushna Journal of Photochemistry and Photobiology C: Photochemistry Reviews, Vol. 12, Issue 1, p. 20-30 https://doi.org/10.1016/j.jphotochemrev.2011.05.001	journal	March 2011
Superlocalization Spectral Imaging Microscopy of a Multicolor Quantum Dot Complex Shi, Xingbo; Xie, Zhongqiu; Song, Yuehong Analytical Chemistry, Vol. 84, Issue 3 https://doi.org/10.1021/ac202784h	journal	January 2012
In vivo hyperspectral confocal fluorescence imaging to determine pigment localization and distribution in cyanobacterial cells Vermaas, W. F. J.; Timlin, J. A.; Jones, H. D. T. Proceedings of the National Academy of Sciences, Vol. 105, Issue 10, p. 4050-4055 https://doi.org/10.1073/pnas.0708090105	journal	March 2008
Hyperspectral confocal microscope Sinclair, Michael B.; Haaland, David M.; Timlin, Jerilyn A. Applied Optics, Vol. 45, Issue 24, p. 6283-6291 https://doi.org/10.1364/AO.45.006283	journal	January 2006
STED microscopy with a supercontinuum laser source Wildanger, Dominik; Rittweger, Eva; Kastrup, Lars Optics Express, Vol. 16, Issue 13 https://doi.org/10.1364/OE.16.009614	journal	January 2008

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