title: Integrated photoacoustic, confocal, and two-photon microscope.
abstract: The invention of green fluorescent protein and other molecular fluorescent probes has promoted applications of confocal and two-photon fluorescence microscopy in biology and medicine. However, exogenous fluorescence contrast agents may affect cellular structure and function, and fluorescence microscopy cannot image nonfluorescent chromophores. We overcome this limitation by integrating optical-resolution photoacoustic microscopy into a modern Olympus IX81 confocal, two-photon, fluorescence microscope setup to provide complementary, label-free, optical absorption contrast. Automatically coregistered images can be generated from the same sample. Imaging applications in ophthalmology, developmental biology, and plant science are demonstrated. For the first time, in a familiar microscopic fluorescence imaging setting, this trimodality microscope provides a platform for future biological and medical discoveries.
authors.affiliation: Washington University in St. Louis, Optical Imaging Laboratory, Department of Biomedical Engineering, One Brookings Drive, St. Louis, Missouri 63130.
authors.forename: Bin
authors.initials: B
authors.lastname: Rao
authors.affiliation: Washington University School of Medicine, Department of Ophthalmology and Visual Sciences, Box 8096, St. Louis, Missouri 63110.
authors.forename: Florentina
authors.initials: F
authors.lastname: Soto
authors.affiliation: Washington University School of Medicine, Department of Ophthalmology and Visual Sciences, Box 8096, St. Louis, Missouri 63110.
authors.forename: Daniel
authors.initials: D
authors.lastname: Kerschensteiner
authors.affiliation: Washington University in St. Louis, Optical Imaging Laboratory, Department of Biomedical Engineering, One Brookings Drive, St. Louis, Missouri 63130.
authors.forename: Lihong V
authors.initials: LV
authors.lastname: Wang
mesh_terms: D000818:Animals; D004625:Embryo, Nonmammalian; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry; D006651:Histocytochemistry; D008164:Luminescent Proteins / Q000032:analysis / Q000737:chemistry; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000379:methods; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation* / Q000379:methods; D061088:Photoacoustic Techniques / Q000295:instrumentation* / Q000379:methods; D012160:Retina; D015027:Zebrafish
chemical_list: D005456:Fluorescent Dyes; D008164:Luminescent Proteins