A long Stokes shift red fluorescent Ca2+ indicator protein for two-photon and ratiometric imaging.
Nat Commun
; 5: 5262, 2014 Oct 31.
Article
em En
| MEDLINE
| ID: mdl-25358432
ABSTRACT
The introduction of calcium ion (Ca(2+)) indicators based on red fluorescent proteins (RFPs) has created new opportunities for multicolour visualization of intracellular Ca(2+) dynamics. However, one drawback of these indicators is that they have optimal two-photon excitation outside the near-infrared window (650-1,000 nm) where tissue is most transparent to light. To address this shortcoming, we developed a long Stokes shift RFP-based Ca(2+) indicator, REX-GECO1, with optimal two-photon excitation at <1,000 nm. REX-GECO1 fluoresces at 585 nm when excited at 480 nm or 910 nm by a one- or two-photon process, respectively. We demonstrate that REX-GECO1 can be used as either a ratiometric or intensiometric Ca(2+) indicator in organotypic hippocampal slice cultures (one- and two-photon) and the visual system of albino tadpoles (two-photon). Furthermore, we demonstrate single excitation wavelength two-colour Ca(2+) and glutamate imaging in organotypic cultures.
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Base de dados:
MEDLINE
Assunto principal:
Prótons
/
Engenharia de Proteínas
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Cálcio
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Imagem Óptica
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Proteínas Luminescentes
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article