title: Optical properties of mouse brain tissue after optical clearing with FocusClear™.
abstract: Fluorescence microscopy is commonly used to investigate disease progression in biological tissues. Biological tissues, however, are strongly scattering in the visible wavelengths, limiting the application of fluorescence microscopy to superficial (<200µm) regions. Optical clearing, which involves incubation of the tissue in a chemical bath, reduces the optical scattering in tissue, resulting in increased tissue transparency and optical imaging depth. The goal of this study was to determine the time- and wavelength-resolved dynamics of the optical scattering properties of rodent brain after optical clearing with FocusClear™. Light transmittance and reflectance of 1-mm mouse brain sections were measured using an integrating sphere before and after optical clearing and the inverse adding doubling algorithm used to determine tissue optical scattering. The degree of optical clearing was quantified by calculating the optical clearing potential (OCP), and the effects of differing OCP were demonstrated using the optical histology method, which combines tissue optical clearing with optical imaging to visualize the microvasculature. We observed increased tissue transparency with longer optical clearing time and an analogous increase in OCP. Furthermore, OCP did not vary substantially between 400 and 1000 nm for increasing optical clearing durations, suggesting that optical histology can improve ex vivo visualization of several fluorescent probes.
authors.affiliation: University of California, Irvine, Beckman Laser Institute and Medical Clinic, Department of Surgery, 1002 Health Sciences Road East, Irvine, California 92617, United StatesbUniversity of California, Irvine, Department of Biomedical Engineering, 3120 Natur.
authors.forename: Austin J
authors.initials: AJ
authors.lastname: Moy
authors.affiliation: University of California, Irvine, Beckman Laser Institute and Medical Clinic, Department of Surgery, 1002 Health Sciences Road East, Irvine, California 92617, United States.
authors.forename: Bernard V
authors.initials: BV
authors.lastname: Capulong
authors.affiliation: University of California, Irvine, Beckman Laser Institute and Medical Clinic, Department of Surgery, 1002 Health Sciences Road East, Irvine, California 92617, United States.
authors.forename: Rolf B
authors.initials: RB
authors.lastname: Saager
authors.affiliation: University of California, Irvine, Beckman Laser Institute and Medical Clinic, Department of Surgery, 1002 Health Sciences Road East, Irvine, California 92617, United StatesbUniversity of California, Irvine, Department of Biomedical Engineering, 3120 Natur.
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Wiersma
authors.affiliation: University of California, Irvine, Beckman Laser Institute and Medical Clinic, Department of Surgery, 1002 Health Sciences Road East, Irvine, California 92617, United StatesbUniversity of California, Irvine, Department of Biomedical Engineering, 3120 Natur.
authors.forename: Patrick C
authors.initials: PC
authors.lastname: Lo
authors.affiliation: University of California, Irvine, Beckman Laser Institute and Medical Clinic, Department of Surgery, 1002 Health Sciences Road East, Irvine, California 92617, United States.
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Durkin
authors.affiliation: University of California, Irvine, Beckman Laser Institute and Medical Clinic, Department of Surgery, 1002 Health Sciences Road East, Irvine, California 92617, United StatesbUniversity of California, Irvine, Department of Biomedical Engineering, 3120 Natur.
authors.forename: Bernard
authors.initials: B
authors.lastname: Choi
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology* / Q000502:physiology*; D003198:Computer Simulation; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008027:Light; D051379:Mice; D008809:Mice, Inbred C3H; D008856:Microscopy, Fluorescence / Q000379:methods*; D008954:Models, Biological; D009924:Organ Culture Techniques; D019188:Organ Preservation Solutions / Q000737:chemistry*; D012542:Scattering, Radiation; D012996:Solutions
chemical_list: D019188:Organ Preservation Solutions; D012996:Solutions