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Un versátil microscopio en miniatura de dos fotones que permite imágenes multicolores del cerebro profundo

Runlong Wu1,2,3, Chunzhu Zhao4,5, Shan Qiu6

  • 1National Biomedical Imaging Center, State Key Laboratory of Membrane Biology, Institute of Molecular Medicine, Peking-Tsinghua Center for Life Sciences, College of Future Technology, Peking University, Beijing, China. rlwu@bistu.edu.cn.

Nature methods
|August 21, 2025
PubMed
Resumen
Este resumen es generado por máquina.

Desarrollamos el FHIRM-TPM 3.0, un microscopio en miniatura para imágenes del cerebro profundo en ratones. Este avanzado sistema de microscopía de dos fotones permite imágenes multicolores de la actividad neuronal y las estructuras celulares in vivo.

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Área de la Ciencia:

  • La neurociencia
  • Ingeniería biomédica
  • Tecnología de microscopía

Sus antecedentes:

  • Las imágenes cerebrales profundas en animales que se comportan libremente son cruciales para entender los circuitos neuronales.
  • Los sistemas de microscopía de dos fotones existentes se enfrentan a limitaciones en tamaño, flexibilidad y profundidad de imagen.
  • Las imágenes multicolores son esenciales para diseccionar procesos celulares y moleculares complejos en el cerebro.

Objetivo del estudio:

  • Para presentar el FHIRM-TPM 3.0, un nuevo y compacto microscopio de dos fotones para imágenes avanzadas del cerebro in vivo.
  • Para demostrar la capacidad del sistema para imágenes multicolores del cerebro profundo en ratones de comportamiento libre.
  • Mostrar la versatilidad y la alta resolución del sistema para diversas aplicaciones de investigación en neurociencias.

Principales métodos:

  • Integración de un microscopio de dos fotones en miniatura con una fibra hueca antirresonante de banda ancha.
  • Corrección de las aberraciones ópticas y optimización de la recolección de fluorescencia para la penetración en tejidos profundos.
  • Ingeniería de objetivos intercambiables para lograr campos de visión escalables y alta resolución lateral.
  • Utilizando longitudes de onda de excitación multicolor (780, 920, 1030 nm) para el monitoreo simultáneo de la actividad celular.

Principales resultados:

  • Obtención de imágenes neuronales corticales a profundidades superiores a 820 μm.
  • Habilitado imágenes de Ca2+ en el hipocampo con una única resolución de columna dendrítica utilizando una lente GRIN.
  • Proporciona un campo de visión escalable diez veces (hasta 1 × 0,8 mm2) con resoluciones desde 0,68 μm hasta 1,46 μm.
  • Se han investigado con éxito las actividades mitocondriales y citosólicas de Ca2+ en relación con las placas amiloides en ratones APP/PS1.

Conclusiones:

  • FHIRM-TPM 3.0 es una herramienta versátil y poderosa para la obtención de imágenes cerebrales profundas multicolores en la investigación de las neurociencias.
  • La miniaturización del sistema y el diseño óptico avanzado facilitan los estudios in vivo en sujetos de comportamiento libre.
  • Permite imágenes de alta resolución de tejidos profundos, avanzando en el estudio de las enfermedades neurológicas y la función cerebral.