Segmentation and 3D reconstruction of microtubules in total internal reflection fluorescence microscopy (TIRFM)

Abstract

The interaction of the microtubules with the cell cortex plays numerous critical roles in a cell. For instance, it directs vesicle delivery, and modulates membrane adhesions pivotal for cell movement as well as mitosis. Abnormal function of the microtubules is involved in cancer. An effective method to observe microtubule function adjacent to the cortex is TIRFM. To date most analysis of TIRFM images has been done by visual inspection and manual tracing. In this work we have developed a method to automatically process TIRFM images of microtubules so as to enable high throughput quantitative studies. The microtubules are extracted in terms of consecutive segments. The segments are described via Hamilton-Jacobi equations. Subsequently, the algorithm performs a limited reconstruction of the microtubules in 3D. Last, we evaluate our method with phantom as well as real TIRFM images of living cells.