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Spatial Molecular Imaging of the Glycome Using Mass Spectrometry
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De la microdisección láser a la glicómica espacial: protocolos de microarrays de lectinas para el mapeo del glicoma
Patcharaporn Boottanun1,2, Sayaka Fuseya1, Atsushi Kuno1
1Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan.
Methods in molecular biology (Clifton, N.J.)
|February 2, 2026
Resumen
La microdisección láser combinada con microarray de lectinas (LMD-LMA) perfila con precisión la glicosilación espacial en tejidos fijados en formalina e incrustados en parafina (FFPE). Este método permite el mapeo detallado del glicoma tisular para una mejor interpretación biológica en ómicas espaciales.
Área de la Ciencia:
- Bioquímica
- Biología Molecular
- Histología
Sus antecedentes:
- Los tejidos fijados en formalina e incrustados en parafina (FFPE) están ampliamente disponibles y conservan la arquitectura tisular.
- La glicómica espacial requiere el aislamiento preciso de regiones tisulares a microescala.
- La microdisección láser (LMD) y el microarray de lectinas (LMA) son tecnologías clave para el perfilado glucómico.
Objetivo del estudio:
- Presentar protocolos detallados para la integración de LMD con LMA (LMD-LMA) para el perfilado glucómico espacial.
- Optimizar el flujo de trabajo LMD-LMA utilizando un escáner avanzado de fluorescencia de campo evanescente.
- Permitir el perfilado glucómico preciso y reproducible de regiones tisulares a microescala.
Principales métodos:
- Se desarrollaron tres enfoques LMD-LMA: guiado por morfología, guiado por sonda en secciones seriadas y disección directa de secciones teñidas.
- Se optimizó el flujo de trabajo con un escáner de fluorescencia de campo evanescente de alta gama (GSR2300) para una mayor sensibilidad y un menor ruido.
- Se estandarizó la canalización de LMA: extracción de proteínas, marcaje con Cy3, interacción lectina-glicoproteína, escaneo de fluorescencia y calificación de datos.
Principales resultados:
- Se logró un perfilado glucómico preciso de regiones tisulares a microescala (aprox. 0.1 mm², 5 μm de espesor).
- Se revelaron patrones de glicosilación intra e inter-tisulares específicos.
- Se generaron conjuntos de datos "mapeo del glicoma tisular" compartibles a través de LM-GlycoRepo y se visualizaron a través de LM-GlycomeAtlas bajo los principios FAIR.
Conclusiones:
- LMD-LMA proporciona un protocolo "plug-and-play" para la glicómica espacial, compatible con las canalizaciones LMD existentes.
- Permite conjuntos de datos co-registrados para transcriptómica y proteómica espacial al reflejar las regiones microdiseccionadas.
- Fomenta la integración de la información de glicanos en flujos de trabajo de ómicas espaciales más amplios para una mejor interpretación biológica.
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