Evidence map›Paper›PMID 42763863›Full record

ArticleMethods in molecular biology (Clifton, N.J.)2027

Monitoring Calcium Dynamics in Pig Intestinal Organoids with FLIM and FRET.

Michele Cervellera, Hang Zhou, Bram van den Broek, Kees Jalink, Ruslan I Dmitriev

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Article in Methods in molecular biology (Clifton, N.J.), 2027. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it
0cells of the map it votes in
0citing papers in PubMed
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1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

5 authors.

Michele CervelleraTissue Engineering and Biomaterials Group, Department of Human Structure and Repair, Faculty of Medicine and Health Sciences, Ghent University, The Core, C. Heymanslaan 10, Ghent, 9000, Belgium.
Hang ZhouTissue Engineering and Biomaterials Group, Department of Human Structure and Repair, Faculty of Medicine and Health Sciences, Ghent University, The Core, C. Heymanslaan 10, Ghent, 9000, Belgium.
Bram van den BroekDivision of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, Amsterdam, 1066CX, The Netherlands.
Kees JalinkDivision of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, Amsterdam, 1066CX, The Netherlands.
Ruslan I DmitrievTissue Engineering and Biomaterials Group, Department of Human Structure and Repair, Faculty of Medicine and Health Sciences, Ghent University, The Core, C. Heymanslaan 10, Ghent, 9000, Belgium. Ruslan.dmitriev@ugent.be.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Intracellular calcium signaling plays a crucial role in intestinal epithelial function. In this chapter, we describe a method to monitor calcium dynamics in live pig small intestinal organoids using stably expressed genetically encoded FRET-based biosensor in combination with intensity-based ratiometric and fluorescence lifetime imaging (FLIM) microscopy readouts. The calcium biosensor Twitch-2B was introduced into organoids by electroporation. Despite the modest lifetime contrast, the biosensor provides a highly reliable ratiometric signal and is compatible with both conventional fluorescence and FLIM-based imaging platform. Following electroporation, organoids are allowed to form and subsequently were selected to establish stable biosensor-expressing lines. This is followed by the live microscopy of heterogeneous and cell-specific calcium responses upon pharmacological stimulation. The presented protocol offers a versatile approach for studies of calcium signaling in relevant 3D cell models, such as organoids and tissues-on-a-chip.

Indexed as

Biosensing TechniquesCalciumCalcium SignalingFluorescence Resonance Energy TransferIntestinesOptical ImagingOrganoidsAnimalsElectroporationMicroscopy, FluorescenceSwineCalciumCalcium signalingElectroporationFluorescence lifetime imaging microscopy (FLIM)Förster resonance energy transfer (FRET)Intestinal organoidsLive cell imaging

Identifiers

PMID42763863

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.