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Fluoróforos de triarilmetano resistentes al fotoazulamiento oxidativo

Alexey N Butkevich^1,2, Mariano L Bossi², Gražvydas Lukinavičius¹

¹Department of NanoBiophotonics , Max Planck Institute for Biophysical Chemistry , Am Fassberg 11 , 37077 Göttingen , Germany.

Journal of the American Chemical Society

|December 19, 2018

Resumen

Desarrollamos nuevos fluoróforos de rodamina con estabilidad espectral mejorada para imágenes multicolores. Estas sondas resisten el foto blanqueamiento y la fotoconversión, mejorando la reproducibilidad de los datos en microscopía de súper resolución.

Área de la Ciencia:

Síntesis química y fotofísica
Microscopía y espectroscopia de fluorescencia
Ciencias de los materiales para sondas de imágenes

Sus antecedentes:

La estabilidad espectral de las sondas fluorescentes es crucial para obtener imágenes de fluorescencia multicolor precisas.
Las altas intensidades de luz en la microscopía de superresolución desafían la estabilidad de la sonda, afectando la reproducibilidad de los datos.
El foto blanqueamiento y la fotoconversión son las principales limitaciones de los fluoróforos actuales.

Objetivo del estudio:

Para sintetizar fluoróforos de rodamina espectralmente estables.
Evaluar su fotoestabilidad frente al fotoblanqueamiento y la fotoconversión.
Para demostrar su utilidad en la microscopía de super-resolución de células vivas.

Principales métodos:

Síntesis secuencial catalizada por Ru y Cu de nuevos derivados de la rodamina.
Análisis quimiométrico para comparar la fotostabilidad con los fluoróforos existentes.
La nanoscopia con células vivas STED (agotamiento estimulado de las emisiones) para evaluar el rendimiento.

Principales resultados:

Se han identificado patrones de sustitución para los fluoróforos de triarilmetano resistentes a la fotoconversión.
Se ha demostrado una fotoestabilidad superior y resistencia al fotoblanqueamiento y a la fotoconversión.
Aplicó con éxito las etiquetas no azules en la nanoscopia STED de células vivas.

Conclusiones:

Desarrolló una ruta sintética para los fluoróforos de rodamina altamente fotostables.
Estas nuevas sondas ofrecen una estabilidad espectral mejorada para aplicaciones de imágenes exigentes.
Las etiquetas no azules son adecuadas para imágenes de células vivas de alta resolución.

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