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J Biophotonics. 2014 Jun;7(6):425-33. doi: 10.1002/jbio.201200175. Epub 2012 Nov 26.

Real-time interactive two-photon photoconversion of recirculating lymphocytes for discontinuous cell tracking in live adult mice.

Author information

1
Immunological Diseases Division, Garvan Institute of Medical Research, Sydney, Australia; St Vincent's Clinical School, Faculty of Medicine, University of New South Wales, Sydney, Australia. t.chtanova@garvan.org.au.

Abstract

The potential usefulness of intravital two-photon microscopy for fate mapping is limited by its inability to track cells beyond the confines of the imaging volume. Therefore, we have developed and validated a novel method for in vivo photolabelling of spatially-restricted cells expressing the Kaede optical highlighter by two-photon excitation. This has allowed us to optically mark a cohort of follicular B cells and track their dissemination from the original imaging volume in the lymph node to the spleen and contralateral lymph node. We also present the first demonstration, to our knowledge, of in vivo photoconversion of a freely moving single cell in a live adult animal. This method of `discontinuous' cell tracking therefore significantly extends the fate mapping capabilities of two-photon microscopy to delineate the spatiotemporal dynamics of cellular processes that span multiple anatomical sites at the single cell level.

KEYWORDS:

Kaede protein; cell tracking; multiphoton fluorescence microscopy; optics and photonics

PMID:
23184395
DOI:
10.1002/jbio.201200175
[Indexed for MEDLINE]

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