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Tracking Single Cells in Live Animals Using a Photoconvertible Near-Infrared Cell Membrane Label

We describe a novel photoconversion technique to track individual cells in vivo using a commercial lipophilic membrane dye, DiR. We show that DiR exhibits a permanent fluorescence emission shift (photoconversion) after light exposure and does not reacquire the original color over time. Ratiometric i...

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Autores principales: Carlson, Alicia L., Fujisaki, Joji, Wu, Juwell, Runnels, Judith M., Turcotte, Raphaël, Celso, Cristina Lo, Scadden, David T., Strom, Terry B., Lin, Charles P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3753322/
https://www.ncbi.nlm.nih.gov/pubmed/23990881
http://dx.doi.org/10.1371/journal.pone.0069257
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author Carlson, Alicia L.
Fujisaki, Joji
Wu, Juwell
Runnels, Judith M.
Turcotte, Raphaël
Celso, Cristina Lo
Scadden, David T.
Strom, Terry B.
Lin, Charles P.
author_facet Carlson, Alicia L.
Fujisaki, Joji
Wu, Juwell
Runnels, Judith M.
Turcotte, Raphaël
Celso, Cristina Lo
Scadden, David T.
Strom, Terry B.
Lin, Charles P.
author_sort Carlson, Alicia L.
collection PubMed
description We describe a novel photoconversion technique to track individual cells in vivo using a commercial lipophilic membrane dye, DiR. We show that DiR exhibits a permanent fluorescence emission shift (photoconversion) after light exposure and does not reacquire the original color over time. Ratiometric imaging can be used to distinguish photoconverted from non-converted cells with high sensitivity. Combining the use of this photoconvertible dye with intravital microscopy, we tracked the division of individual hematopoietic stem/progenitor cells within the calvarium bone marrow of live mice. We also studied the peripheral differentiation of individual T cells by tracking the gain or loss of FoxP3-GFP expression, a marker of the immune suppressive function of CD4(+) T cells. With the near-infrared photoconvertible membrane dye, the entire visible spectral range is available for simultaneous use with other fluorescent proteins to monitor gene expression or to trace cell lineage commitment in vivo with high spatial and temporal resolution.
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spelling pubmed-37533222013-08-29 Tracking Single Cells in Live Animals Using a Photoconvertible Near-Infrared Cell Membrane Label Carlson, Alicia L. Fujisaki, Joji Wu, Juwell Runnels, Judith M. Turcotte, Raphaël Celso, Cristina Lo Scadden, David T. Strom, Terry B. Lin, Charles P. PLoS One Research Article We describe a novel photoconversion technique to track individual cells in vivo using a commercial lipophilic membrane dye, DiR. We show that DiR exhibits a permanent fluorescence emission shift (photoconversion) after light exposure and does not reacquire the original color over time. Ratiometric imaging can be used to distinguish photoconverted from non-converted cells with high sensitivity. Combining the use of this photoconvertible dye with intravital microscopy, we tracked the division of individual hematopoietic stem/progenitor cells within the calvarium bone marrow of live mice. We also studied the peripheral differentiation of individual T cells by tracking the gain or loss of FoxP3-GFP expression, a marker of the immune suppressive function of CD4(+) T cells. With the near-infrared photoconvertible membrane dye, the entire visible spectral range is available for simultaneous use with other fluorescent proteins to monitor gene expression or to trace cell lineage commitment in vivo with high spatial and temporal resolution. Public Library of Science 2013-08-26 /pmc/articles/PMC3753322/ /pubmed/23990881 http://dx.doi.org/10.1371/journal.pone.0069257 Text en © 2013 Carlson et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Carlson, Alicia L.
Fujisaki, Joji
Wu, Juwell
Runnels, Judith M.
Turcotte, Raphaël
Celso, Cristina Lo
Scadden, David T.
Strom, Terry B.
Lin, Charles P.
Tracking Single Cells in Live Animals Using a Photoconvertible Near-Infrared Cell Membrane Label
title Tracking Single Cells in Live Animals Using a Photoconvertible Near-Infrared Cell Membrane Label
title_full Tracking Single Cells in Live Animals Using a Photoconvertible Near-Infrared Cell Membrane Label
title_fullStr Tracking Single Cells in Live Animals Using a Photoconvertible Near-Infrared Cell Membrane Label
title_full_unstemmed Tracking Single Cells in Live Animals Using a Photoconvertible Near-Infrared Cell Membrane Label
title_short Tracking Single Cells in Live Animals Using a Photoconvertible Near-Infrared Cell Membrane Label
title_sort tracking single cells in live animals using a photoconvertible near-infrared cell membrane label
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3753322/
https://www.ncbi.nlm.nih.gov/pubmed/23990881
http://dx.doi.org/10.1371/journal.pone.0069257
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