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Proceedings Paper

Large field of view quantitative phase imaging of induced pluripotent stem cells and optical pathlength reference materials
Author(s): Edward Kwee; Alexander Peterson; Jeffrey Stinson; Michael Halter; Liya Yu; Michael Majurski; Joe Chalfoun; Peter Bajcsy; John Elliott
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Paper Abstract

Induced pluripotent stem cells (iPSCs) are reprogrammed cells that can have heterogeneous biological potential. Quality assurance metrics of reprogrammed iPSCs will be critical to ensure reliable use in cell therapies and personalized diagnostic tests. We present a quantitative phase imaging (QPI) workflow which includes acquisition, processing, and stitching multiple adjacent image tiles across a large field of view (LFOV) of a culture vessel.

Low magnification image tiles (10x) were acquired with a Phasics SID4BIO camera on a Zeiss microscope. iPSC cultures were maintained using a custom stage incubator on an automated stage. We implement an image acquisition strategy that compensates for non-flat illumination wavefronts to enable imaging of an entire well plate, including the meniscus region normally obscured in Zernike phase contrast imaging. Polynomial fitting and background mode correction was implemented to enable comparability and stitching between multiple tiles. LFOV imaging of reference materials indicated that image acquisition and processing strategies did not affect quantitative phase measurements across the LFOV. Analysis of iPSC colony images demonstrated mass doubling time was significantly different than area doubling time.

These measurements were benchmarked with prototype microsphere beads and etched-glass gratings with specified spatial dimensions designed to be QPI reference materials with optical pathlength shifts suitable for cell microscopy.

This QPI workflow and the use of reference materials can provide non-destructive traceable imaging method for novel iPSC heterogeneity characterization.

Paper Details

Date Published: 23 February 2018
PDF: 8 pages
Proc. SPIE 10503, Quantitative Phase Imaging IV, 105032F (23 February 2018); doi: 10.1117/12.2290691
Show Author Affiliations
Edward Kwee, National Institute of Standards and Technology (United States)
Alexander Peterson, National Institute of Standards and Technology (United States)
Jeffrey Stinson, National Institute of Standards and Technology (United States)
Michael Halter, National Institute of Standards and Technology (United States)
Liya Yu, National Institute of Standards and Technology (United States)
Michael Majurski, National Institute of Standards and Technology (United States)
Joe Chalfoun, National Institute of Standards and Technology (United States)
Peter Bajcsy, National Institute of Standards and Technology (United States)
John Elliott, National Institute of Standards and Technology (United States)


Published in SPIE Proceedings Vol. 10503:
Quantitative Phase Imaging IV
Gabriel Popescu; YongKeun Park, Editor(s)

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