Forward-collected simultaneous fluorescence lifetime imaging and coherent anti-Stokes Raman scattering microscopy

Aaron D. Slepkov; Andrew Ridsdale; Huei Ning Wan; Ming Hao Wang; Adrian F. Pegoraro; Douglas J. Moffatt; John P. Pezacki; Fu Jen Kao; Albert Stolow

doi:10.1117/1.3490641

Forward-collected simultaneous fluorescence lifetime imaging and coherent anti-Stokes Raman scattering microscopy

Aaron D. Slepkov, Andrew Ridsdale, Huei Ning Wan, Ming Hao Wang, Adrian F. Pegoraro, Douglas J. Moffatt, John P. Pezacki, Fu Jen Kao, Albert Stolow

Institute of Biophotonics

Research output: Contribution to journal › Article › peer-review

20 Scopus citations

Abstract

We demonstrate the simultaneous collection and separation of femtosecond-laser-based forward-collected coherent anti-Stokes Raman scattering (F-CARS) and two-photon-excitation-induced fluorescence lifetime images (FLIM) using time-correlated single photon counting (TCSPC). We achieve this in a nondescanned geometry using a single multimode fiber without significant loss of light, field of view, and most importantly, TCSPC timing fidelity. In addition to showing the ability to separate CARS images from FLIM images using time gating, we also demonstrate composite multimodal epicollected FLIM imaging with fiber-collected F-CARS imaging in live cells.

Original language	English
Article number	021103
Journal	Journal of Biomedical Optics
Volume	16
Issue number	2
DOIs	https://doi.org/10.1117/1.3490641
State	Published - Feb 2011

Keywords

Raman spectroscopy
fiber optic applications
laser-induced fluorescence
nonlinear optics
scanning microscopy

Access to Document

10.1117/1.3490641

Cite this

@article{040595fe75fb47b99a1387f610aa6cb7,

title = "Forward-collected simultaneous fluorescence lifetime imaging and coherent anti-Stokes Raman scattering microscopy",

abstract = "We demonstrate the simultaneous collection and separation of femtosecond-laser-based forward-collected coherent anti-Stokes Raman scattering (F-CARS) and two-photon-excitation-induced fluorescence lifetime images (FLIM) using time-correlated single photon counting (TCSPC). We achieve this in a nondescanned geometry using a single multimode fiber without significant loss of light, field of view, and most importantly, TCSPC timing fidelity. In addition to showing the ability to separate CARS images from FLIM images using time gating, we also demonstrate composite multimodal epicollected FLIM imaging with fiber-collected F-CARS imaging in live cells.",

keywords = "Raman spectroscopy, fiber optic applications, laser-induced fluorescence, nonlinear optics, scanning microscopy",

author = "Slepkov, {Aaron D.} and Andrew Ridsdale and Wan, {Huei Ning} and Wang, {Ming Hao} and Pegoraro, {Adrian F.} and Moffatt, {Douglas J.} and Pezacki, {John P.} and Kao, {Fu Jen} and Albert Stolow",

note = "Funding Information: This research was largely funded by a joint NRC-NSC-ITRI Canada-Taiwan Cooperation project “Digital FLIM-CARS Microscopy.” We would like to thank Rodney Lyn for his expertise and time in HuH-7 cell sample preparation and general discussion. We gratefully acknowledge Guillaume Labroille for fruitful discussions and for his input into optical alignment and layout design.",

year = "2011",

month = feb,

doi = "10.1117/1.3490641",

language = "English",

volume = "16",

journal = "Journal of Biomedical Optics",

issn = "1083-3668",

publisher = "SPIE",

number = "2",

}

TY - JOUR

T1 - Forward-collected simultaneous fluorescence lifetime imaging and coherent anti-Stokes Raman scattering microscopy

AU - Slepkov, Aaron D.

AU - Ridsdale, Andrew

AU - Wan, Huei Ning

AU - Wang, Ming Hao

AU - Pegoraro, Adrian F.

AU - Moffatt, Douglas J.

AU - Pezacki, John P.

AU - Kao, Fu Jen

AU - Stolow, Albert

N1 - Funding Information: This research was largely funded by a joint NRC-NSC-ITRI Canada-Taiwan Cooperation project “Digital FLIM-CARS Microscopy.” We would like to thank Rodney Lyn for his expertise and time in HuH-7 cell sample preparation and general discussion. We gratefully acknowledge Guillaume Labroille for fruitful discussions and for his input into optical alignment and layout design.

PY - 2011/2

Y1 - 2011/2

N2 - We demonstrate the simultaneous collection and separation of femtosecond-laser-based forward-collected coherent anti-Stokes Raman scattering (F-CARS) and two-photon-excitation-induced fluorescence lifetime images (FLIM) using time-correlated single photon counting (TCSPC). We achieve this in a nondescanned geometry using a single multimode fiber without significant loss of light, field of view, and most importantly, TCSPC timing fidelity. In addition to showing the ability to separate CARS images from FLIM images using time gating, we also demonstrate composite multimodal epicollected FLIM imaging with fiber-collected F-CARS imaging in live cells.

AB - We demonstrate the simultaneous collection and separation of femtosecond-laser-based forward-collected coherent anti-Stokes Raman scattering (F-CARS) and two-photon-excitation-induced fluorescence lifetime images (FLIM) using time-correlated single photon counting (TCSPC). We achieve this in a nondescanned geometry using a single multimode fiber without significant loss of light, field of view, and most importantly, TCSPC timing fidelity. In addition to showing the ability to separate CARS images from FLIM images using time gating, we also demonstrate composite multimodal epicollected FLIM imaging with fiber-collected F-CARS imaging in live cells.

KW - Raman spectroscopy

KW - fiber optic applications

KW - laser-induced fluorescence

KW - nonlinear optics

KW - scanning microscopy

UR - http://www.scopus.com/inward/record.url?scp=79551714481&partnerID=8YFLogxK

U2 - 10.1117/1.3490641

DO - 10.1117/1.3490641

M3 - Article

C2 - 21361666

AN - SCOPUS:79551714481

SN - 1083-3668

VL - 16

JO - Journal of Biomedical Optics

JF - Journal of Biomedical Optics

IS - 2

M1 - 021103

ER -

Forward-collected simultaneous fluorescence lifetime imaging and coherent anti-Stokes Raman scattering microscopy

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this