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Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein
|Title:||Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein|
Haynes, Russell D.
Corbel, Stéphane Y.
González- González, Emilio
show 12 moreBurg, John S.
Ataie, Niloufar J.
Lam, Amy J.
Cranfill, Paula J.
Baird, Michelle A.
Davidson, Michael W.
Ng, Ho Leung
Garcia, K. Christopher
Contag, Christopher H.
Blau, Helen M.
Lin, Michael Z.
|Date Issued:||16 Mar 2014|
|Citation:||Chu J, Haynes RD, Corbel SY, Li P, González-González E, Burg JS, Ataie NJ, Lam AJ, Cranfill PJ, Baird MA, Davidson MW, Ng HL, Garcia KC, Contag CH, Shen K, Blau HM, Lin MZ. Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein. Nat. Methods 2014;11:572-578.|
|Abstract:||A method for non-invasive visualization of genetically labelled cells in animal disease
models with micron-level resolution would greatly facilitate development of cell-based
therapies. Imaging of fluorescent proteins (FPs) using red excitation light in the “optical
window” above 600 nm is one potential method for visualizing implanted cells. However,
previous efforts to engineer FPs with peak excitation beyond 600 nm have resulted in
undesirable reductions in brightness. Here we report three new red-excitable monomeric FPs obtained by structure-guided mutagenesis of mNeptune, previously the brightest monomeric FP when excited beyond 600 nm. Two of these, mNeptune2 and mNeptune2.5, demonstrate improved maturation and brighter fluorescence, while the third, mCardinal, has a red-shifted excitation spectrum without reduction in brightness. We show that mCardinal can be used to non-invasively and longitudinally visualize the differentiation of myoblasts and stem cells into myocytes in living mice with high anatomical detail.
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Ng, Ho Leung|
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