Large-scale Time-lapse Microscopy of Oct4 Expression in Human Embryonic Stem Cell Colonies

Published

May 22, 2016

Author(s)

Kiran Bhadriraju, Michael Halter, Julien M. Amelot, Peter Bajcsy, Joe Chalfoun, Antoine Vandecreme, Barbara Mallon, Kye-yoon Park, John T. Elliott, Subhash Sista, Anne L. Plant

Abstract

Identification of critical characteristics of therapeutically active cells, and quantification of those characteristics, are essential for assuring consistency and potency of cell therapy products. We have developed image analysis and visualization software that allows effective use of time-lapse microscopy to provide spatial and dynamic information from large numbers of human embryonic stem cell colonies. To achieve statistically relevant sampling, we examined more than 680 colonies from 3 different preparations of cells over 5 days, generating a total experimental dataset of 0.9 terabyte (TB). The 0.5 Giga-pixel images at each time point were represented by multi-resolution pyramids and visualized using the Deep Zoom Javascript library extended to support viewing Giga-pixel images over time and extracting data on individual colonies. We present a methodology that enables quantification of variations in nominally-identical preparations and between colonies, correlation of colony characteristics with Oct4 expression, and identification of rare events.

Citation

Nature Biotechnology

Volume

17

Pub Type

Journals

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DOI Link

Keywords

Fluorescence microscopy, stem cells, live cell imaging, cell therapy, pluripotency, quantitative optical microscopy

Citation

Bhadriraju, K. , Halter, M. , Amelot, J. , Bajcsy, P. , Chalfoun, J. , Vandecreme, A. , Mallon, B. , Park, K. , Elliott, J. , Sista, S. and Plant, A. (2016), Large-scale Time-lapse Microscopy of Oct4 Expression in Human Embryonic Stem Cell Colonies, Nature Biotechnology, [online], https://doi.org/10.1016/j.scr.2016.05.012 (Accessed November 21, 2024)