Interpretation of Fluctuations in the Basal Cell Membrane while Active Membrane Trafficking

Maji, Debanjana (2017) Interpretation of Fluctuations in the Basal Cell Membrane while Active Membrane Trafficking. Masters thesis, Indian Institute of Science Education and Research Kolkata.

PDF (MS dissertation of Debanjana Maji (12MS038)) Master's_thesis_Debanjana_Maji.pdf - Submitted Version Restricted to Repository staff only Download (3MB)

Abstract

A simple lipid bilayer in solution is subject to thermal excitation and fluctuates incessantly around its mean shape. From these fluctuations, distinct characteristics like correlation length and amplitude can be obtained, which yields important information about the membrane mechanical properties and its interaction with the environment. Membrane fluctuations are obtained by converting intensity maps into height maps and calculating the standard deviation of these heights over time. Previous studies in the lab have identified that there are regions in cell membrane where transient high fluctuations are observed, termed as hotspots. These hotspots are depicted as high intensity values in First Branch Regions (FBRs) (regions, comprising an area 144 pixels, found between a maximum and a minimum intensity spots of an intensity map of a cell). These hotspots appear in the same regions of the FBRs at a time interval of one second and disappear within one second. We hypothesize that these hotspots may occur because of the formation of membrane pits, endosomes (both early and late endosomes) or interaction of vesicles/ Golgi/ Endoplasmic Reticulum with plasma membrane. Therefore, we aim to correlate membrane trafficking with fluctuation hotspots in adhered cells using Interference Reflection Microscopy. To correlate the late events of endocytosis with the formation of hotspots of the cells I have used pHrodo Dextran and to correlate early events of endocytosis with hotspot formation, clathrin transfection has been used.

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