Fluorescent protein-cell labeling and its application in time-lapse analysis of hematopoietic differentiation.
Here, we present a computer-controlled time-lapse system for imaging of cultured hematopoietic cells labeled by the expression of different fluorescent proteins. First, we describe experiments to optimize the visualization of three green fluorescent protein variants (cyan-, green-, and yellow-enhanced fluorescent protein) and the red-fluorescent protein (DsRed) by standard wide-field fluorescence microscopy. Then, we describe procedures to best distinguish combinations of cells expressing these proteins using seven commercially available filter sets, based on the relative fluorescence intensities of the individual fluorescent proteins. Finally, we make recommendations about which of these filters to choose when working with specific fluorescent proteins.
Stadtfeld
Matthias
M
Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, NY, USA.
Varas
Florencio
F
Graf
Thomas
T
D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*
D049452:Green Fluorescent Proteins