Large-scale time-lapse microscopy of Oct4 expression in human embryonic stem cell colonies
Identification and quantification of the characteristics of stem cell preparations is critical for understanding stem cell biology and for the development and manufacturing of stem cell based therapies. We have developed image analysis and visualization software that allows effective use of time-lap...
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doaj-a0e1fb69e8e54282870d293ea35fca752020-11-24T22:26:47ZengElsevierStem Cell Research1873-50611876-77532016-07-0117112212910.1016/j.scr.2016.05.012Large-scale time-lapse microscopy of Oct4 expression in human embryonic stem cell coloniesKiran Bhadriraju0Michael Halter1Julien Amelot2Peter Bajcsy3Joe Chalfoun4Antoine Vandecreme5Barbara S. Mallon6Kye-yoon Park7Subhash Sista8John T. Elliott9Anne L. Plant10Department of Bioengineering, University of Maryland, College Park, MD 20742, USABiosystems and Biomaterials Division, Materials Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899, USASoftware Systems Division, Information Technology Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899, USASoftware Systems Division, Information Technology Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899, USASoftware Systems Division, Information Technology Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899, USASoftware Systems Division, Information Technology Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899, USAThe NIH Stem Cell Unit, Division of Intramural Research, National Institute of Neurological Disorders and Stroke, NIH, U.S. Department of Health and Human Services, Bethesda, MD, USAThe NIH Stem Cell Unit, Division of Intramural Research, National Institute of Neurological Disorders and Stroke, NIH, U.S. Department of Health and Human Services, Bethesda, MD, USABiosystems and Biomaterials Division, Materials Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899, USABiosystems and Biomaterials Division, Materials Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899, USABiosystems and Biomaterials Division, Materials Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899, USAIdentification and quantification of the characteristics of stem cell preparations is critical for understanding stem cell biology and for the development and manufacturing of stem cell based therapies. 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 >680 colonies from 3 different preparations of cells over 5 days each, 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.http://www.sciencedirect.com/science/article/pii/S1873506116300502Fluorescence microscopyStem cellsLive cell imagingCell therapyPluripotency |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Kiran Bhadriraju Michael Halter Julien Amelot Peter Bajcsy Joe Chalfoun Antoine Vandecreme Barbara S. Mallon Kye-yoon Park Subhash Sista John T. Elliott Anne L. Plant |
spellingShingle |
Kiran Bhadriraju Michael Halter Julien Amelot Peter Bajcsy Joe Chalfoun Antoine Vandecreme Barbara S. Mallon Kye-yoon Park Subhash Sista John T. Elliott Anne L. Plant Large-scale time-lapse microscopy of Oct4 expression in human embryonic stem cell colonies Stem Cell Research Fluorescence microscopy Stem cells Live cell imaging Cell therapy Pluripotency |
author_facet |
Kiran Bhadriraju Michael Halter Julien Amelot Peter Bajcsy Joe Chalfoun Antoine Vandecreme Barbara S. Mallon Kye-yoon Park Subhash Sista John T. Elliott Anne L. Plant |
author_sort |
Kiran Bhadriraju |
title |
Large-scale time-lapse microscopy of Oct4 expression in human embryonic stem cell colonies |
title_short |
Large-scale time-lapse microscopy of Oct4 expression in human embryonic stem cell colonies |
title_full |
Large-scale time-lapse microscopy of Oct4 expression in human embryonic stem cell colonies |
title_fullStr |
Large-scale time-lapse microscopy of Oct4 expression in human embryonic stem cell colonies |
title_full_unstemmed |
Large-scale time-lapse microscopy of Oct4 expression in human embryonic stem cell colonies |
title_sort |
large-scale time-lapse microscopy of oct4 expression in human embryonic stem cell colonies |
publisher |
Elsevier |
series |
Stem Cell Research |
issn |
1873-5061 1876-7753 |
publishDate |
2016-07-01 |
description |
Identification and quantification of the characteristics of stem cell preparations is critical for understanding stem cell biology and for the development and manufacturing of stem cell based therapies. 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 >680 colonies from 3 different preparations of cells over 5 days each, 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. |
topic |
Fluorescence microscopy Stem cells Live cell imaging Cell therapy Pluripotency |
url |
http://www.sciencedirect.com/science/article/pii/S1873506116300502 |
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