A Parallel Perifusion Slide From Glass for the Functional and Morphological Analysis of Pancreatic Islets.
NAD(P)H- and FAD-autofluorescence
borosilicate glass
calcium
femtosecond laser-structuring
insulin secretion
islet of langerhans
microfluidic perifusion system
Journal
Frontiers in bioengineering and biotechnology
ISSN: 2296-4185
Titre abrégé: Front Bioeng Biotechnol
Pays: Switzerland
ID NLM: 101632513
Informations de publication
Date de publication:
2021
2021
Historique:
received:
09
10
2020
accepted:
12
02
2021
entrez:
25
3
2021
pubmed:
26
3
2021
medline:
26
3
2021
Statut:
epublish
Résumé
An islet-on-chip system in the form of a completely transparent microscope slide optically accessible from both sides was developed. It is made from laser-structured borosilicate glass and enables the parallel perifusion of five microchannels, each containing one islet precisely immobilized in a pyramidal well. The islets can be in inserted via separate loading windows above each pyramidal well. This design enables a gentle, fast and targeted insertion of the islets and a reliable retention in the well while at the same time permitting a sufficiently fast exchange of the media. In addition to the measurement of the hormone content in the fractionated efflux, parallel live cell imaging of the islet is possible. By programmable movement of the microscopic stage imaging of five wells can be performed. The current chip design ensures sufficient time resolution to characterize typical parameters of stimulus-secretion coupling. This was demonstrated by measuring the reaction of the islets to stimulation by glucose and potassium depolarization. After the perifusion experiment islets can be removed for further analysis. The live-dead assay of the removed islets confirmed that the process of insertion and removal was not detrimental to islet structure and viability. In conclusion, the present islet-on-chip design permits the practical implementation of parallel perifusion experiments on a single and easy to load glass slide. For each immobilized islet the correlation between secretion, signal transduction and morphology is possible. The slide concept allows the scale-up to even higher degrees of parallelization.
Identifiants
pubmed: 33763408
doi: 10.3389/fbioe.2021.615639
pmc: PMC7982818
doi:
Types de publication
Journal Article
Langues
eng
Pagination
615639Informations de copyright
Copyright © 2021 Schulze, Mattern, Erfle, Brüning, Scherneck, Dietzel and Rustenbeck.
Déclaration de conflit d'intérêts
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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