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Snapshot 3D tracking of insulin granules in live cells
Author(s): Xiaolei Wang; Xiang Huang; Itay Gdor; Matthew Daddysman; Hannah Yi; Alan Selewa; Theresa Haunold; Mark Hereld; Norbert F. Scherer
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Paper Abstract

Rapid and accurate volumetric imaging remains a challenge, yet has the potential to enhance understanding of cell function. We developed and used a multifocal microscope (MFM) for 3D snapshot imaging to allow 3D tracking of insulin granules labeled with mCherry in MIN6 cells. MFM employs a special diffractive optical element (DOE) to simultaneously image multiple focal planes. This simultaneous acquisition of information determines the 3D location of single objects at a speed only limited by the array detector’s frame rate. We validated the accuracy of MFM imaging/tracking with fluorescence beads; the 3D positions and trajectories of single fluorescence beads can be determined accurately over a wide range of spatial and temporal scales. The 3D positions and trajectories of single insulin granules in a 3.2um deep volume were determined with imaging processing that combines 3D decovolution, shift correction, and finally tracking using the Imaris software package. We find that the motion of the granules is superdiffusive, but less so in 3D than 2D for cells grown on coverslip surfaces, suggesting an anisotropy in the cytoskeleton (e.g. microtubules and action).

Paper Details

Date Published: 23 February 2018
PDF: 7 pages
Proc. SPIE 10499, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV, 104990R (23 February 2018); doi: 10.1117/12.2285530
Show Author Affiliations
Xiaolei Wang, The Univ. of Chicago (United States)
Xiang Huang, Argonne National Lab. (United States)
Itay Gdor, The Univ. of Chicago (United States)
Matthew Daddysman, The Univ. of Chicago (United States)
Hannah Yi, The Univ. of Chicago (United States)
Alan Selewa, The Univ. of Chicago (United States)
Theresa Haunold, The Univ. of Chicago (United States)
Mark Hereld, Argonne National Lab. (United States)
Norbert F. Scherer, The Univ. of Chicago (United States)


Published in SPIE Proceedings Vol. 10499:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV
Thomas G. Brown; Carol J. Cogswell; Tony Wilson, Editor(s)

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