23 February 2018 Large field of view quantitative phase imaging of induced pluripotent stem cells and optical pathlength reference materials
Author Affiliations +
Proceedings Volume 10503, Quantitative Phase Imaging IV; 105032F (2018) https://doi.org/10.1117/12.2290691
Event: SPIE BiOS, 2018, San Francisco, California, United States
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.
© (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Edward Kwee, Edward Kwee, Alexander Peterson, Alexander Peterson, Jeffrey Stinson, Jeffrey Stinson, Michael Halter, Michael Halter, Liya Yu, Liya Yu, Michael Majurski, Michael Majurski, Joe Chalfoun, Joe Chalfoun, Peter Bajcsy, Peter Bajcsy, John Elliott, John Elliott, } "Large field of view quantitative phase imaging of induced pluripotent stem cells and optical pathlength reference materials", Proc. SPIE 10503, Quantitative Phase Imaging IV, 105032F (23 February 2018); doi: 10.1117/12.2290691; https://doi.org/10.1117/12.2290691

Back to Top