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Extended voltage imaging in cardiomyocytes with a triplet state quencher-stabilized silicon rhodamine.
Martinez, Kayli N; Gerstner, Nels C; Yang, Samantha J; Miller, Evan W.
Afiliación
  • Martinez KN; Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720-1460, USA.
  • Gerstner NC; Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720-1460, USA.
  • Yang SJ; Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720-1460, USA.
  • Miller EW; Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720-1460, USA; Department of Molecular & Cell Biology, University of California, Berkeley, Berkeley, CA 94720-1460, USA; Helen Wills Neuroscience Institute, University of California, Berkeley, Berkeley, CA 94720-1460, US
Bioorg Med Chem Lett ; 109: 129842, 2024 Sep 01.
Article en En | MEDLINE | ID: mdl-38844174
ABSTRACT
Voltage imaging of cardiac electrophysiology with voltage-sensitive dyes has long been a powerful complement to traditional methods like patch-clamp electrophysiology. Chemically synthesized voltage sensitive fluorophores offer flexibility for imaging in sensitive samples like human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs), since they do not require genetic transformation of the sample. One serious concern for any fluorescent voltage indicator, whether chemically synthesized or genetically encoded, is phototoxicity. We have been exploring self-healing fluorophores that use triplet state quenchers (TSQs) as a means to reduce the already low phototoxicity of VoltageFluor dyes developed in our lab. We previously showed that conjugation of the TSQ cyclooctatetraene (COT) to a fluorescein based VoltageFluor dye substantially reduced phototoxicity. Here, we show that this approach can be applied to far-red Silicon rhodamine dyes. COT-conjugated Si-rhodamines show improved photostability and reduced phototoxicity in hiPSC-CMs compared to the unmodified dye. This enables imaging of hiPSC-CMs for up to 30 min with continuous illumination. We show that this effect is mediated by a combination of reduced singlet oxygen production and lower loading in the cellular membrane. We discuss future applications and avenues of improvement for TSQ-stabilized VoltageFluor dyes.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Rodaminas / Miocitos Cardíacos / Células Madre Pluripotentes Inducidas / Colorantes Fluorescentes Límite: Humans Idioma: En Revista: Bioorg Med Chem Lett Asunto de la revista: BIOQUIMICA / QUIMICA Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Rodaminas / Miocitos Cardíacos / Células Madre Pluripotentes Inducidas / Colorantes Fluorescentes Límite: Humans Idioma: En Revista: Bioorg Med Chem Lett Asunto de la revista: BIOQUIMICA / QUIMICA Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido