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Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein.
Chu, Jun; Haynes, Russell D; Corbel, Stéphane Y; Li, Pengpeng; González-González, Emilio; Burg, 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; Shen, Kang; Blau, Helen M; Lin, Michael Z.
Afiliação
  • Chu J; 1] Department of Bioengineering, Stanford University, Stanford, California, USA. [2] Department of Pediatrics, Stanford University School of Medicine, Stanford, California, USA.
  • Haynes RD; 1] Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, USA. [2] Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California, USA.
  • Corbel SY; 1] Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, USA. [2] Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California, USA.
  • Li P; Department of Biological Sciences, Stanford University, Stanford, California, USA.
  • González-González E; 1] Department of Pediatrics, Stanford University School of Medicine, Stanford, California, USA. [2] Molecular Imaging Program at Stanford, Stanford University School of Medicine, Stanford, California, USA.
  • Burg JS; 1] Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California, USA. [2] Department of Structural Biology, Stanford University School of Medicine, Stanford, California, USA.
  • Ataie NJ; Department of Chemistry, University of Hawaii at Manoa, Honolulu, Hawaii, USA.
  • Lam AJ; 1] Department of Bioengineering, Stanford University, Stanford, California, USA. [2] Department of Pediatrics, Stanford University School of Medicine, Stanford, California, USA.
  • Cranfill PJ; 1] Department of Biological Science, Florida State University, Tallahassee, Florida, USA. [2] National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
  • Baird MA; 1] Department of Biological Science, Florida State University, Tallahassee, Florida, USA. [2] National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
  • Davidson MW; 1] Department of Biological Science, Florida State University, Tallahassee, Florida, USA. [2] National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida, USA.
  • Ng HL; 1] Department of Chemistry, University of Hawaii at Manoa, Honolulu, Hawaii, USA. [2] University of Hawaii Cancer Center, Honolulu, Hawaii, USA.
  • Garcia KC; 1] Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California, USA. [2] Department of Structural Biology, Stanford University School of Medicine, Stanford, California, USA. [3] Howard Hughes Medical Institute, Stanford University, Stanford, Californ
  • Contag CH; 1] Department of Pediatrics, Stanford University School of Medicine, Stanford, California, USA. [2] Molecular Imaging Program at Stanford, Stanford University School of Medicine, Stanford, California, USA.
  • Shen K; 1] Department of Biological Sciences, Stanford University, Stanford, California, USA. [2] Howard Hughes Medical Institute, Stanford University, Stanford, California, USA.
  • Blau HM; 1] Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, USA. [2] Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California, USA.
  • Lin MZ; 1] Department of Bioengineering, Stanford University, Stanford, California, USA. [2] Department of Pediatrics, Stanford University School of Medicine, Stanford, California, USA. [3] Molecular Imaging Program at Stanford, Stanford University School of Medicine, Stanford, California, USA.
Nat Methods ; 11(5): 572-8, 2014 May.
Article em En | MEDLINE | ID: mdl-24633408
ABSTRACT
A method for non-invasive visualization of genetically labeled cells in animal disease models with micrometer-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. Two of these, mNeptune2 and mNeptune2.5, demonstrate improved maturation and brighter fluorescence than mNeptune, whereas 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 into myocytes in living mice with high anatomical detail.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Diagnóstico por Imagem / Diferenciação Celular / Proteínas Luminescentes / Microscopia de Fluorescência Tipo de estudo: Diagnostic_studies Limite: Animals / Humans / Male Idioma: En Ano de publicação: 2014 Tipo de documento: Article
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Diagnóstico por Imagem / Diferenciação Celular / Proteínas Luminescentes / Microscopia de Fluorescência Tipo de estudo: Diagnostic_studies Limite: Animals / Humans / Male Idioma: En Ano de publicação: 2014 Tipo de documento: Article