NDLI: In vivo clonal tracking of hematopoietic stem and progenitor cells marked by five fluorescent proteins using confocal and multiphoton microscopy.

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In vivo clonal tracking of hematopoietic stem and progenitor cells marked by five fluorescent proteins using confocal and multiphoton microscopy.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Malide, Daniela Métais, Jean-Yves Dunbar, Cynthia E.
Description	Author Affiliation: Malide D ( Light Microscopy Core Facility, NHLBI/NIH); Métais JY ( Hematology Branch, NHLBI/NIH.); Dunbar CE ( Hematology Branch, NHLBI/NIH.)
Abstract	We developed and validated a fluorescent marking methodology for clonal tracking of hematopoietic stem and progenitor cells (HSPCs) with high spatial and temporal resolution to study in vivo hematopoiesis using the murine bone marrow transplant experimental model. Genetic combinatorial marking using lentiviral vectors encoding fluorescent proteins (FPs) enabled cell fate mapping through advanced microscopy imaging. Vectors encoding five different FPs: Cerulean, EGFP, Venus, tdTomato, and mCherry were used to concurrently transduce HSPCs, creating a diverse palette of color marked cells. Imaging using confocal/two-photon hybrid microscopy enables simultaneous high resolution assessment of uniquely marked cells and their progeny in conjunction with structural components of the tissues. Volumetric analyses over large areas reveal that spectrally coded HSPC-derived cells can be detected non-invasively in various intact tissues, including the bone marrow (BM), for extensive periods of time following transplantation. Live studies combining video-rate multiphoton and confocal time-lapse imaging in 4D demonstrate the possibility of dynamic cellular and clonal tracking in a quantitative manner.
File Format	HTM / HTML
e-ISSN	1940087X
DOI	10.3791/51669
Journal	Journal of Visualized Experiments
Issue Number	90
Language	English
Publisher	MyJove Corp.
Publisher Date	2014-08-06
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine Cell Tracking Fluorescent Antibody Technique Hematopoietic Stem Cells Chemistry Cytology Microscopy, Confocal Microscopy, Fluorescence, Multiphoton Animals Clone Cells Hematopoietic Stem Cell Transplantation Luminescent Proteins Genetics Mice Mice, Inbred C57bl Research Support, N.i.h., Intramural Video-audio Media
Content Type	Text
Resource Type	Article

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