Two-color, two-photon imaging at long excitation wavelengths using a diamond Raman laser

Johanna Tragardh; Michelle Murtagh; Gillian Robb; Maddy Parsons; Jipeng Lin; David J. Spence; Gail McConnell

doi:10.1017/S143192761601151X

Two-color, two-photon imaging at long excitation wavelengths using a diamond Raman laser

Johanna Tragardh, Michelle Murtagh, Gillian Robb, Maddy Parsons, Jipeng Lin, David J. Spence, Gail McConnell

Strathclyde Institute Of Pharmacy And Biomedical Sciences

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

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Abstract

We demonstrate that the second-Stokes output from a diamond Raman laser, pumped by a femtosecond Ti:Sapphire laser, can be used to efficiently excite red-emitting dyes by two-photon excitation at 1080 nm and beyond. We image red fluorescent protein (RFP) expressing HeLa cells, as well as dyes such as Texas Red and Mitotracker Red. We demonstrate the potential for simultaneous two-color, two-photon imaging with this laser by using the residual pump beam for excitation of a green-emitting dye. We demonstrate this for the combination of AlexaFluor 488 and AlexaFluor 568. Because the Raman laser extends the wavelength range of the Ti:Sapphire laser, resulting in a laser system tunable 680 nm-1200 nm it can be used for two-photon excitation of a large variety and combination of dyes.

Original language	English
Number of pages	5
Journal	Microscopy and Microanalysis
Early online date	5 Aug 2016
DOIs	https://doi.org/10.1017/S143192761601151X
Publication status	E-pub ahead of print - 5 Aug 2016

Keywords

nonlinear microscopy
fluorescence microscopy
raman laser
fluorescent proteins
two-photon microscopy

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10.1017/S143192761601151X

Tragardh-etal-MM-2016-two-color-two-photon-imaging-at-long-excitation-wavelengthsAccepted author manuscript, 322 KBLicence: Unspecified

Cite this

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title = "Two-color, two-photon imaging at long excitation wavelengths using a diamond Raman laser",

abstract = "We demonstrate that the second-Stokes output from a diamond Raman laser, pumped by a femtosecond Ti:Sapphire laser, can be used to efficiently excite red-emitting dyes by two-photon excitation at 1080 nm and beyond. We image red fluorescent protein (RFP) expressing HeLa cells, as well as dyes such as Texas Red and Mitotracker Red. We demonstrate the potential for simultaneous two-color, two-photon imaging with this laser by using the residual pump beam for excitation of a green-emitting dye. We demonstrate this for the combination of AlexaFluor 488 and AlexaFluor 568. Because the Raman laser extends the wavelength range of the Ti:Sapphire laser, resulting in a laser system tunable 680 nm-1200 nm it can be used for two-photon excitation of a large variety and combination of dyes.",

keywords = "nonlinear microscopy, fluorescence microscopy, raman laser , fluorescent proteins, two-photon microscopy",

author = "Johanna Tragardh and Michelle Murtagh and Gillian Robb and Maddy Parsons and Jipeng Lin and Spence, {David J.} and Gail McConnell",

year = "2016",

month = aug,

day = "5",

doi = "10.1017/S143192761601151X",

language = "English",

journal = "Microscopy and Microanalysis",

issn = "1431-9276",

publisher = "Cambridge University Press",

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TY - JOUR

T1 - Two-color, two-photon imaging at long excitation wavelengths using a diamond Raman laser

AU - Tragardh, Johanna

AU - Murtagh, Michelle

AU - Robb, Gillian

AU - Parsons, Maddy

AU - Lin, Jipeng

AU - Spence, David J.

AU - McConnell, Gail

PY - 2016/8/5

Y1 - 2016/8/5

N2 - We demonstrate that the second-Stokes output from a diamond Raman laser, pumped by a femtosecond Ti:Sapphire laser, can be used to efficiently excite red-emitting dyes by two-photon excitation at 1080 nm and beyond. We image red fluorescent protein (RFP) expressing HeLa cells, as well as dyes such as Texas Red and Mitotracker Red. We demonstrate the potential for simultaneous two-color, two-photon imaging with this laser by using the residual pump beam for excitation of a green-emitting dye. We demonstrate this for the combination of AlexaFluor 488 and AlexaFluor 568. Because the Raman laser extends the wavelength range of the Ti:Sapphire laser, resulting in a laser system tunable 680 nm-1200 nm it can be used for two-photon excitation of a large variety and combination of dyes.

AB - We demonstrate that the second-Stokes output from a diamond Raman laser, pumped by a femtosecond Ti:Sapphire laser, can be used to efficiently excite red-emitting dyes by two-photon excitation at 1080 nm and beyond. We image red fluorescent protein (RFP) expressing HeLa cells, as well as dyes such as Texas Red and Mitotracker Red. We demonstrate the potential for simultaneous two-color, two-photon imaging with this laser by using the residual pump beam for excitation of a green-emitting dye. We demonstrate this for the combination of AlexaFluor 488 and AlexaFluor 568. Because the Raman laser extends the wavelength range of the Ti:Sapphire laser, resulting in a laser system tunable 680 nm-1200 nm it can be used for two-photon excitation of a large variety and combination of dyes.

KW - nonlinear microscopy

KW - fluorescence microscopy

KW - raman laser

KW - fluorescent proteins

KW - two-photon microscopy

UR - http://journals.cambridge.org/jid_MAM

U2 - 10.1017/S143192761601151X

DO - 10.1017/S143192761601151X

M3 - Article

SN - 1431-9276

JO - Microscopy and Microanalysis

JF - Microscopy and Microanalysis

ER -

Two-color, two-photon imaging at long excitation wavelengths using a diamond Raman laser

Abstract

Keywords

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MESOLAB: A Centre For Optical Mesoscopy For Biomedical Research At The University Of Strathclyde

Versatile Ultrafast Raman Laser Sources For Biophotonics - Macquarie University