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Label-free photoacoustic microscopy of cytochromes

Zhang, Chi and Zhang, Yu Shrike and Yao, Da-Kang and Xia, Younan and Wang, Lihong V. (2013) Label-free photoacoustic microscopy of cytochromes. Journal of Biomedical Optics, 18 (2). Art. No. 020504. ISSN 1083-3668. PMCID PMC3560837.

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Photoacoustic microscopy (PAM) has achieved submicron lateral resolution in showing subcellular structures; however, relatively few endogenous subcellular contrasts have so far been imaged. Given that the hemeprotein, mostly cytochromes in general cells, is optically absorbing around the Soret peak (∼420  nm), we implemented label-free PAM of cytochromes in cytoplasm for the first time. By measuring the photoacoustic spectra of the oxidized and reduced states of fibroblast lysate and fitting the difference spectrum with three types of cytochromes, we found that the three cytochromes account for more than half the optical absorption in the cell lysate at 420 nm wavelength. Fixed fibroblasts on slides were imaged by PAM at 422 and 250 nm wavelengths to reveal cytoplasms and nuclei, respectively, as confirmed by standard staining histology. PAM was also applied to label-free histology of mouse ear sections by showing cytoplasms and nuclei of various cells. PAM of cytochromes in cytoplasm is expected to be a high-throughput, label-free technique for studying live cell functions, which cannot be accomplished by conventional histology.

Item Type:Article
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URLURL TypeDescription CentralArticle
Wang, Lihong V.0000-0001-9783-4383
Additional Information:© 2013 SPIE. Paper 12680LRR received Oct. 16, 2012; revised manuscript received Jan. 15, 2013; accepted for publication Jan. 16, 2013; published online Jan. 31, 2013. This work was sponsored in part by National Institutes of Health (NIH) grants R01 EB000712, R01 EB008085, R01 CA134539, U54 CA136398, R01 CA157277, and R01 CA159959. L.W. has a financial interest in Microphotoacoustics Inc. and Endra Inc., which did not support this work.
Funding AgencyGrant Number
NIHR01 EB000712
NIHR01 EB008085
NIHR01 CA134539
NIHU54 CA136398
NIHR01 CA157277
NIHR01 CA159959
Subject Keywords:photoacoustic microscopy; photoacoustic spectroscopy; cytochrome; mitochondria; label-free
PubMed Central ID:PMC3560837
Record Number:CaltechAUTHORS:20160803-111418126
Persistent URL:
Official Citation:Zhang C, Zhang Y, Yao D, Xia Y, Wang LV; Label-free photoacoustic microscopy of cytochromes. J. Biomed. Opt. 0001;18(2):020504-020504. doi:10.1117/1.JBO.18.2.020504
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:69410
Deposited By: Tony Diaz
Deposited On:03 Aug 2016 23:53
Last Modified:03 Aug 2016 23:53

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