title: Two-photonin vivoflow cytometry using a fiber probe.
abstract: We have demonstrated the use of a double-clad fiber probe to conduct two-photon excited flow cytometryin vitroandin vivo. We conducted two-channel detection to measure fluorescence at two distinct wavelengths simultaneously. Because the scattering and absorption problems from whole blood were circumvented by the fiber probe, the detected signal strength from the cells were found to be similar in PBS and in whole blood. We achieved the same detection efficiency of the membrane-binding lipophilic dye DiD labeled cells in PBS and in whole blood. High detection efficiency of green fluorescent protein (GFP)-expressing cells in whole blood was demonstrated. DiD-labeled untransfected and GFP-transfected cells were injected into live mice and the circulation dynamics of the externally injected cells were monitored. The detection efficiency of GFP-expressing cellsin vivowas consistent with that observed in whole blood.
authors.affiliation: Center for Ultrafast Optical Science, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Yu-Chung
authors.initials: YC
authors.lastname: Chang
authors.affiliation: Center for Ultrafast Optical Science, University of Michigan, Ann Arbor, MI 48109; Michigan Nanotechnology Institute for Medicine and Biological Sciences, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Jing Yong
authors.initials: JY
authors.lastname: Ye
authors.affiliation: Michigan Nanotechnology Institute for Medicine and Biological Sciences, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Thommey P
authors.initials: TP
authors.lastname: Thomas
authors.affiliation: Michigan Nanotechnology Institute for Medicine and Biological Sciences, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Zhengyi
authors.initials: Z
authors.lastname: Cao
authors.affiliation: Michigan Nanotechnology Institute for Medicine and Biological Sciences, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Alina
authors.initials: A
authors.lastname: Kotlyar
authors.affiliation: Center for Ultrafast Optical Science, University of Michigan, Ann Arbor, MI 48109; Michigan Nanotechnology Institute for Medicine and Biological Sciences, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Eric R
authors.initials: ER
authors.lastname: Tkaczyk
authors.affiliation: Michigan Nanotechnology Institute for Medicine and Biological Sciences, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109.
authors.forename: James R
authors.initials: JR
authors.lastname: Baker
authors.affiliation: Center for Ultrafast Optical Science, University of Michigan, Ann Arbor, MI 48109; Michigan Nanotechnology Institute for Medicine and Biological Sciences, Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109.
authors.forename: Theodore B
authors.initials: TB
authors.lastname: Norris
keywords: Two-photon fluorescence; cancer; circulating tumor cell; double-clad fiber; fiber probe; green fluorescent protein (GFP); in vivo flow cytometry; metastasis