Journal
BIOMEDICAL OPTICS EXPRESS
Volume 4, Issue 11, Pages 2477-2490Publisher
OPTICAL SOC AMER
DOI: 10.1364/BOE.4.002477
Keywords
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Funding
- King's College London and University College London Comprehensive Cancer Imaging Centre Cancer Research UK
- UK Engineering and Physical Sciences Research Council
- European Research Council
- BBSRC [BB/J018899/1] Funding Source: UKRI
- EPSRC [EP/H005536/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/J018899/1] Funding Source: researchfish
- Engineering and Physical Sciences Research Council [EP/H005536/1] Funding Source: researchfish
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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. (C) 2013 Optical Society of America
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