title: Longitudinal label-free tracking of cell death dynamics in living engineered human skin tissue with a multimodal microscope.
abstract: We demonstrate real-time, longitudinal, label-free tracking of apoptotic and necrotic cells in living tissue using a multimodal microscope. The integrated imaging platform combines multi-photon microscopy (MPM, based on two-photon excitation fluorescence), optical coherence microscopy (OCM), and fluorescence lifetime imaging microscopy (FLIM). Three-dimensional (3-D) co-registered images are captured that carry comprehensive information of the sample, including structural, molecular, and metabolic properties, based on light scattering, autofluorescence intensity, and autofluorescence lifetime, respectively. Different cell death processes, namely, apoptosis and necrosis, of keratinocytes from different epidermal layers are longitudinally monitored and investigated. Differentiation of the two cell death processes in a complex living tissue environment is enabled by quantitative image analysis and high-confidence classification processing based on the multidimensional, cross-validating imaging data. These results suggest that despite the limitations of each individual label-free modality, this multimodal imaging approach holds the promise for studies of different cell death processes in living tissue and in vivo organs.
authors.affiliation: Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Youbo
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Marina
authors.initials: M
authors.lastname: Marjanovic
authors.affiliation: Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Chaney
authors.affiliation: Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA ; Department of Electrical and Computer Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Benedikt W
authors.initials: BW
authors.lastname: Graf
authors.affiliation: Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA ; Department of Kinesiology and Community Health, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Ziad
authors.initials: Z
authors.lastname: Mahmassani
authors.affiliation: Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA ; Department of Kinesiology and Community Health, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Marni D
authors.initials: MD
authors.lastname: Boppart
authors.affiliation: Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA ; Department of Electrical and Computer Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA ; Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA ; Department of Internal Medicine, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Boppart
keywords: (170.1530) Cell analysis; (170.1870) Dermatology; (170.4500) Optical coherence tomography; (180.2520) Fluorescence microscopy; (180.4315) Nonlinear microscopy