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

Research output: Contribution to journalArticle

1 Citation (Scopus)

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.
LanguageEnglish
Number of pages5
JournalMicroscopy and Microanalysis
Early online date5 Aug 2016
DOIs
Publication statusE-pub ahead of print - 5 Aug 2016

Fingerprint

Diamond
Raman lasers
Photons
Diamonds
Lasers
Color
dyes
diamonds
color
Imaging techniques
Wavelength
photons
Coloring Agents
Dyes
wavelengths
excitation
lasers
sapphire
Aluminum Oxide
Sapphire

Keywords

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

Cite this

Tragardh, Johanna ; Murtagh, Michelle ; Robb, Gillian ; Parsons, Maddy ; Lin, Jipeng ; Spence, David J. ; McConnell, Gail. / Two-color, two-photon imaging at long excitation wavelengths using a diamond Raman laser. In: Microscopy and Microanalysis. 2016.
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Two-color, two-photon imaging at long excitation wavelengths using a diamond Raman laser. / Tragardh, Johanna; Murtagh, Michelle; Robb, Gillian; Parsons, Maddy; Lin, Jipeng; Spence, David J.; McConnell, Gail.

In: Microscopy and Microanalysis, 05.08.2016.

Research output: Contribution to journalArticle

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AU - Tragardh, Johanna

AU - Murtagh, Michelle

AU - Robb, Gillian

AU - Parsons, Maddy

AU - Lin, Jipeng

AU - Spence, David J.

AU - McConnell, Gail

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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.

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KW - raman laser

KW - fluorescent proteins

KW - two-photon microscopy

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