3D widefield light microscope image reconstruction without dyes

S. Larkin, J. Larson, C. Holmes, M. Vaicik, M. Turturro, A. Jurkevich, S. Sinha, T. Ezashi, G. Papavasiliou, E. Brey, T. Holmes

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review


3D image reconstruction using light microscope modalities without exogenous contrast agents is proposed and investigated as an approach to produce 3D images of biological samples for live imaging applications. Multimodality and multispectral imaging, used in concert with this 3D optical sectioning approach is also proposed as a way to further produce contrast that could be specific to components in the sample. The methods avoid usage of contrast agents. Contrast agents, such as fluorescent or absorbing dyes, can be toxic to cells or alter cell behavior. Current modes of producing 3D image sets from a light microscope, such as 3D deconvolution algorithms and confocal microscopy generally require contrast agents. Zernike phase contrast (ZPC), transmitted light brightfield (TLB), darkfield microscopy and others can produce contrast without dyes. Some of these modalities have not previously benefitted from 3D image reconstruction algorithms, however. The 3D image reconstruction algorithm is based on an underlying physical model of scattering potential, expressed as the sample's 3D absorption and phase quantities. The algorithm is based upon optimizing an objective function - the I-divergence-while solving for the 3D absorption and phase quantities. Unlike typical deconvolution algorithms, each microscope modality, such as ZPC or TLB, produces two output image sets instead of one. Contrast in the displayed image and 3D renderings is further enabled by treating the multispectral/multimodal data as a feature set in a mathematical formulation that uses the principal component method of statistics.

Original languageEnglish
Title of host publicationThree-Dimensional and Multidimensional Microscopy
Subtitle of host publicationImage Acquisition and Processing XXII
EditorsThomas G. Brown, Carol J. Cogswell, Tony Wilson
ISBN (Electronic)9781628414202
StatePublished - 2015
Externally publishedYes
EventThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXII - San Francisco, United States
Duration: 9 Feb 201512 Feb 2015

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
ISSN (Print)1605-7422


ConferenceThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXII
Country/TerritoryUnited States
CitySan Francisco


  • image reconstruction
  • light microscopy
  • live imaging
  • phase contrast
  • phase imaging
  • quantitative phase imaging
  • three dimensional


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