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Development of a Tool for Visualization of Cross-Correlation of Intercellular Calcium Signaling in MDCK-II Cell Clusters
KTH, School of Engineering Sciences (SCI), Applied Physics.
2025 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesisAlternative title
Utveckling av ett verktyg för visualisering av kalciumsignalkorrelationer mellan MDCK-II-celler i kluster (Swedish)
Abstract [en]

Intercellular calcium waves are implicated in a multitude of signaling processes in the body. They have been found to influence both wound healing and cancer tumor growth. Understanding signaling patterns of calcium waves could therefore yield important new insights which could lead to improved therapeutical methods.

This thesis develops a tool for finding and visually representing crosscorrelations between intracellular calcium signals of cells in a cluster, as a means of representing communication between cells through intercellular calcium waves. Madin-Darby Canine Kidney II (MDCK II) cells expressing the calcium indicator GCaMP6m were imaged using fluorescence microscopy time-lapse imaging. Images were subsequently analyzed using previously developed tools based on Cellpose 2.0 to perform segmentation, tracking and calcium signal extraction. Finally, a tool was created that calculated cross-correlation of intracellular calcium signals, evaluated significance, and mapped correlations onto an image of the cell cluster.

The results of cross-correlation analysis indicated some patterns of interest in how cells within the imaged clusters communicate. In all clusters, oneor two cells had correlations with all other cells in the network showing correlations above the significance threshold, whereas other cells only had one or a few cross-correlations. Results indicate that the tool developed could be useful in revealing intercellular calcium signaling patterns.

Abstract [sv]

Intercellulära kalciumvågor utgör en mekanism för intercellulär kalciumsignalering som styr en mängd processer i kroppen. De har konstaterats påverka såväl sårläkning som tillväxt hos cancertumörer. Därmed skulle kartläggning av mönster i intercellulär kalciumsignalering kunna leda till viktiga insikter som kan bidra till utveckling av bättre medicinska behandlingsmetoder.

Detta examensarbete syftade till att utveckla en metod för att analysera korrelationen mellan intracellulära kalciumsignaler i olika celler, som ett sätt att modellera kommunikationen mellan celler via intercellulära kalciumvågor. Celler av typen Madin-Darby Canine Kidney II som transfekterats med kalciumindikatorn GCaMP6m avbildades i tidsserier med hjälp av fluorescensmikroskopi. Därefter analyserades bilderna med redan tillgängliga verktyg baserade på Cellpose 2.0 för att segmentera och följa cellerna samt extrahera deras respektive kalciumsignaler. Slutligen skapades ett verktyg för att beräkna korrelationskoefficienterna mellan kalciumsignalerna från olika celler, utvärdera deras signifikans, samt skapa en visuell representation av resultaten ovanpå bilden av cellklustret.

Resultaten av korrelationsanalysen indikerade att det fanns vissa mönster i hur cellerna i klustren signalerade till varandra. I samtliga kluster fanns en ellertvå celler med korrelation med alla andra i det nätverk av celler vars inbördes korrelationer översteg gränsen för signifikans. Samtidigt visade andra celler bara en eller några få signifikanta korrelationer övriga. Resultaten antyder att det utvecklade verktyget skulle kunna användas för att undersöka intercellulära kalciumsignaleringsmönster.

Place, publisher, year, edition, pages
2025.
Series
TRITA-SCI-GRU ; 2025:004
Keywords [en]
Calcium signaling, MDCK II, GCaMP6, Cellpose 2.0, Intercellular calcium waves, Cross-correlation, Visualization
Keywords [sv]
Kalciumsignaliering, MDCK II, GCaMP6, Intercellulära kalciumvågor, Korrelationssamband, Visuell representation
National Category
Physical Sciences
Identifiers
URN: urn:nbn:se:kth:diva-358766OAI: oai:DiVA.org:kth-358766DiVA, id: diva2:1929711
Subject / course
Physics
Educational program
Master of Science - Engineering Physics
Supervisors
Examiners
Available from: 2025-01-21 Created: 2025-01-21 Last updated: 2025-01-21Bibliographically approved

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