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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 4, Iss. 11 — Nov. 1, 2013
  • pp: 2477–2490

In vitro characterization of genetically expressed absorbing proteins using photoacoustic spectroscopy

Jan Laufer, Amit Jathoul, Martin Pule, and Paul Beard  »View Author Affiliations

Biomedical Optics Express, Vol. 4, Issue 11, pp. 2477-2490 (2013)

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Genetically expressed fluorescent proteins have been shown to provide photoacoustic contrast. However, they can be limited by low photoacoustic generation efficiency and low optical absorption at red and near infrared wavelengths, thus limiting their usefulness in mammalian small animal models. In addition, many fluorescent proteins exhibit low photostability due to photobleaching and transient absorption effects. In this study, we explore these issues by synthesizing and characterizing a range of commonly used fluorescent proteins (dsRed, mCherry, mNeptune, mRaspberry, AQ143, E2 Crimson) and novel non-fluorescent chromoproteins (aeCP597 and cjBlue and a non-fluorescent mutant of E2 Crimson). The photoacoustic spectra, photoacoustic generation efficiency and photostability of each fluorescent protein and chromoprotein were measured. Compared to the fluorescent proteins, the chromoproteins were found to exhibit higher photoacoustic generation efficiency due to the absence of radiative relaxation and ground state depopulation, and significantly higher photostability. The feasibility of converting an existing fluorescent protein into a non-fluorescent chromoprotein via mutagenesis was also demonstrated. The chromoprotein mutant exhibited greater photoacoustic signal generation efficiency and better agreement between the photoacoustic and the specific extinction coefficient spectra than the original fluorescent protein. Lastly, the genetic expression of a chromoprotein in mammalian cells was demonstrated. This study suggests that chromoproteins may have potential for providing genetically encoded photoacoustic contrast.

© 2013 Optical Society of America

OCIS Codes
(000.1430) General : Biology and medicine
(300.0300) Spectroscopy : Spectroscopy
(110.5125) Imaging systems : Photoacoustics

ToC Category:
Photoacoustic Imaging and Spectroscopy

Original Manuscript: August 7, 2013
Revised Manuscript: October 1, 2013
Manuscript Accepted: October 1, 2013
Published: October 14, 2013

Virtual Issues
November 22, 2013 Spotlight on Optics

Jan Laufer, Amit Jathoul, Martin Pule, and Paul Beard, "In vitro characterization of genetically expressed absorbing proteins using photoacoustic spectroscopy," Biomed. Opt. Express 4, 2477-2490 (2013)

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