Monitoring real-time hormone release kinetics via high-content 3-D imaging of compensatory endocytosis

Andrei I Tarasov, Juris Galvanovskis, Olof Rorsman, Alexander Hamilton, Elisa Vergari, Paul R V Johnson, Frank Reimann, Frances M Ashcroft, Patrik Rorsman

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskriftPeer review

11 Citeringar (SciVal)

Sammanfattning

High-content real-time imaging of hormone secretion in tissues or cell populations is a challenging task, which is unlikely to be resolved directly, despite immense translational value. We approach this problem indirectly, using compensatory endocytosis, a process that closely follows exocytosis in the cell, as a surrogate read-out for secretion. The tissue is immobilized in an open-air perifusion chamber and imaged using a two-photon microscope. A fluorescent polar tracer, perifused through the experimental circuit, gets trapped into the cells via endocytosis, and is quantified using a feature-detection algorithm. The signal of the tracer that accumulates into the endocytotic system reliably reflects stimulated exocytosis, which is demonstrated via co-imaging of the latter using existing reporters. A high signal-to-noise ratio and compatibility with multisensor imaging affords the real-time quantification of the secretion at the tissue/population level, whereas the cumulative nature of the signal allows imprinting of the "secretory history" within each cell. The technology works for several cell types, reflects disease progression and can be used for human tissue.

Originalspråkengelska
Sidor (från-till)2838-2848
Antal sidor11
TidskriftLab on a Chip
Volym18
Utgåva18
DOI
StatusPublished - 2018 sep. 11
Externt publiceradJa

Ämnesklassifikation (UKÄ)

  • Cell- och molekylärbiologi

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