Journal
PLOS ONE
Volume 8, Issue 8, Pages -Publisher
PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0069257
Keywords
-
Categories
Funding
- National Institutes of Health (NIH) [HL097748, HL97794, CA086355, HL100402]
- NIH [5 PO1 AI041521-12]
- Juvenile Diabetes Research Foundation [7-2005-1329, 4-2007-1057]
- JDRF Center in Immunologic Tolerance
- BBSRC [BB/I004033/1, BB/K021168/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/K021168/1, BB/I004033/1] Funding Source: researchfish
- Medical Research Council [MR/J006742/1] Funding Source: researchfish
Ask authors/readers for more resources
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available