Hinchazón celular y obtención de imágenes basada en montaje vertical para la visualización de alto resolución del...

Área de la Ciencia:

• Cell biology
• Neuroscience
• Biotechnology

Sus antecedentes:

• Receptor-mediated transcytosis (RMT) is key for delivering therapeutics across the blood-brain barrier (BBB).
• Visualizing RMT pathways is challenging due to limitations in current imaging techniques.
• Existing methods often lack resolution or require expensive, complex equipment.

Objetivo del estudio:

• To develop a simplified, high-resolution imaging approach for visualizing RMT.
• To elucidate the intracellular trafficking pathways of RMT cargos.
• To enable direct observation of vesicle dynamics during transcytosis.

Principales métodos:

• Introduced a cell swelling and upright mounting (CSUM)-based imaging approach.
• Utilized standard confocal microscopy to reorient Z-axis into the XY-plane.
• Tracked transferrin (Tf) and anti-transferrin receptor antibody (anti-TfR Ab) using compartment-specific markers and co-localization analysis.

Principales resultados:

• CSUM enabled direct visualization of RMT without computational reconstruction.
• Observed vesicles containing Tf and anti-TfR Ab trafficking from apical to basolateral membranes.
• Tf transcytosis completed within 15 minutes; anti-TfR Ab showed initial endolysosomal pathway entry then rerouting.

Conclusiones:

• The CSUM approach offers a simple, effective platform for high-resolution imaging of membrane transport.
• This method facilitates the study of vesicle dynamics in RMT.
• Provides broad applicability for drug delivery research and designing brain-targeted therapeutics.