AbstractsPhysics

In living cells, calcium ions (Ca2+) play an important role as an intracellular second messenger. It mediates the regulation of cellular processes such as gene expression, initiation of vesicle fusion in synapses, is used in muscle contraction and is believed to play a fundamental role in synaptic plasticity as a molecular substrate for learning. The Ca2+ signals are created by the fact that the concentration of Ca2+ in the cytosol is four orders of magnitude lower than in the extracellular fluid as well as in cytoplasmic compartments such as the endoplasmic reticulum (ER). This enables fast increments in the cytosol concentration, which is regulated back to normal concentration by different mechanisms. In this project, the connection between Ca2+ signals of different cells was analysed using different correlation techniques: cross-correlation of continuous signals and digitalised signals. Therefore a software tool was developed in MATLAB, which takes Ca2+ recordings from time-lapse fluorescence microscopy as input and calculates the pair wise correlation for all cells. The software was tested by using previous data from experiments with embryonic stem cells from mouse (mES) and human (hES) as well as data from recordings done as part of the project. The study shows that the mathematical method of cross-correlation can successfully be applied to quantitative and qualititative analysis of Ca2+ signals. Furthermore, there exist strongly correlated cells in colonies of mES cells and hES cells. We suggest the synchronisation is achieved by physical coupling implicating a decrease of correlation as the distance increases for strong correlations. In addition, the lag used by the cross-correlation function (an effective phase shift) decreases as the correlation coefficient increases and increases as the intercellular distance increases for high correlation coefficients. Interestingly, the number of cells included in small scale clusters of strongly correlated cells is significantly larger for the differentiating mES cells than for the proliferating mÉS cells. In a broader perspective, the developed software might be usd in for instance analysis of cellular electrical activity and shows the relevance of applying methods from the exact sciences to biology.