Mapping the subcellular energy landscape with 4D microscopy

We will develop new methods for real time 3D analysis of living cells – 4D microscopy.

We use two different microscopy techniques – lattice light sheet microscopy (LLSM) and our own construction of multifocus structured illumination (MF-SIM).With those techniques we aim to map the dynamic subcellular energy landscape and explore how mitochondria are transported seemingly random but superimposed by energy gradients.

We hypothesize that the random motion of mitochondria is constitutive and result in an energy optimized subcellular organization.

We will study biological systems with increasing complexity, starting with single cells and moving up to tissue models and organoids.

We seek to identify the decisive moment when an unstructured growing clump of cells is transformed into an ordered structure with a defined inside/out direction.To experimentally address the fundamental biophysical questions we study energy homeostasis in relation to the sodium pump, Na,K-ATPase, the single largest consumer of energy in the body.

We use a cell line from dog kidney that we grow into polarized structures - hollow cysts.

We express fluorescent proteins in the cells to map and follow organelles and proteins of interest.The project is of relevance to fundamental biophysics as well as to human diseases, including genetic disorders affecting mitochondria dynamics and more common hypertension, where the regulation of Na,K-ATPase is of relevance for kidney function and blood pressure.