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Multispectral fingerprinting for improved in vivo cell dynamics analysis

Kulesa, Paul M. and Teddy, Jessica M. and Smith, Miranda and Alexander, Richard and Cooper, Cameron H. J. and Lansford, Rusty and McLennan, Rebecca (2010) Multispectral fingerprinting for improved in vivo cell dynamics analysis. BMC Developmental Biology, 10 . Art. No. 101. ISSN 1471-213X. PMCID PMC2954993. https://resolver.caltech.edu/CaltechAUTHORS:20101103-111543029

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[img] Video (AVI) (Time-lapse analysis reveals that single-colored individual neural crest cells are difficult to follow when cell trajectories cross) - Supplemental Material
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[img] Video (AVI) (Time-lapse analysis reveals that multicolor-labeled neural crest cells are easier to follow when cell trajectories cross) - Supplemental Material
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[img] Video (AVI) (Time-lapse analysis reveals that multicolor-labeled cell movements are traceable within the dense neural tube) - Supplemental Material
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[img] Image (TIFF) (NC Cell Spectral Identity Remains Consistent in Short Term (~5 hr) Time-Lapse Imaging) - Supplemental Material
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Abstract

Background: Tracing cell dynamics in the embryo becomes tremendously difficult when cell trajectories cross in space and time and tissue density obscure individual cell borders. Here, we used the chick neural crest (NC) as a model to test multicolor cell labeling and multispectral confocal imaging strategies to overcome these roadblocks. Results: We found that multicolor nuclear cell labeling and multispectral imaging led to improved resolution of in vivo NC cell identification by providing a unique spectral identity for each cell. NC cell spectral identity allowed for more accurate cell tracking and was consistent during short term time-lapse imaging sessions. Computer model simulations predicted significantly better object counting for increasing cell densities in 3-color compared to 1-color nuclear cell labeling. To better resolve cell contacts, we show that a combination of 2-color membrane and 1-color nuclear cell labeling dramatically improved the semi-automated analysis of NC cell interactions, yet preserved the ability to track cell movements. We also found channel versus lambda scanning of multicolor labeled embryos significantly reduced the time and effort of image acquisition and analysis of large 3D volume data sets. Conclusions: Our results reveal that multicolor cell labeling and multispectral imaging provide a cellular fingerprint that may uniquely determine a cell's position within the embryo. Together, these methods offer a spectral toolbox to resolve in vivo cell dynamics in unprecedented detail.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1186/1471-213X-10-101DOIArticle
http://www.biomedcentral.com/1471-213X/10/101PublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2954993/PubMed CentralArticle
ORCID:
AuthorORCID
Smith, Miranda0000-0002-3321-1432
Lansford, Rusty0000-0002-2159-3699
Additional Information:© 2010 Kulesa et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Received: 28 May 2010 Accepted: 24 September 2010. Published: 24 September 2010. We would like to thank Hua Li for consultation on statistical analysis of data and Winfried Wiegraebe and Joel Schwartz for helpful technical discussions. This work was funded by NIH grant 1R01HD057922 and the Stowers Institute for Medical Research. Authors’ contributions RM, JMT, and MS performed all the embryo labeling, sample preparation, multispectral imaging, and image analysis in the DiI, GFP, and multi-color H2B-labeling experiments and cell tracking. CHJC and JMT performed all the embryo labeling, sample preparation, multispectral imaging, and image analysis in the multi-color cell membrane labeling experiments. RA designed and carried out the computational model simulations. RL provided many of the fluorescent protein constructs and participated in discussions of the experimental design strategy. PMK designed the research, analyzed data and wrote the paper. All authors read and approved the final manuscript.
Funders:
Funding AgencyGrant Number
NIH1R01HD057922
Stowers Institute for Medical ResearchUNSPECIFIED
PubMed Central ID:PMC2954993
Record Number:CaltechAUTHORS:20101103-111543029
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20101103-111543029
Official Citation:Kulesa et al.: Multispectral fingerprinting for improved in vivo cell dynamics analysis. BMC Developmental Biology 2010 10:101.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:20650
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:04 Nov 2010 15:08
Last Modified:09 Mar 2020 13:19

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