Large-scale time-lapse microscopy of Oct4 expression in human embryonic stem cell colonies

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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Main Authors: 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
Format: Article
Language:English
Published: Elsevier 2016-07-01
Series:Stem Cell Research
Subjects:
Fluorescence microscopy
Stem cells
Live cell imaging
Cell therapy
Pluripotency
Online Access:http://www.sciencedirect.com/science/article/pii/S1873506116300502
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spelling 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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