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 journalArticlepeer-review

7 Citations (Scopus)
257 Downloads (Pure)


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


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


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