Receptor Internalization Assay
Name
Taavi Tetlov
Abstract
High-content screening has proved itself to be a highly efficent method for analyzing bioassays in various different fields from oncology to drug screening. Cellomics HCS platform is a modular system which provides the means to analyze cellular and subcellular processes by using fluorescent dyes and automatic fluorescent microscopy.
The aim of this thesis is to describe receptor internalization bioassay in biological system developed by Icosagen. Main focus of the thesis is to describe both image and data analysis methods and processes behind the final description of the assay.
Studied assay is Endothelin A receptor internalization assay and the goal was to investigate the relation of peptide Endothelin-1 and internalization activity. By using Cellomics Colocalization V3 BioApplication for image analysis and fitting four parameter logistic curve on receptor and early-endosome overlap medians it was found that the Endothelin-1 effective doses are 0.29nM for the assay to show any activity, 1.6nM for half-maximal efficency and 10nM for maximum internalization.
As an alternative data analysis method Kolmogorov-Smirnov two sample test for difference in variance was also tested. Results of this approach were fitted to four-parameter logistic model and the sigmoidal curves suggested that this method can be used as an alternative to more common study of mean or median values.
Graduation Thesis language
Estonian
Graduation Thesis type
Bachelor - Information Technology
Supervisor(s)
Raivo Kolde, Andres Tover ja Sven Laur
Defence year
2012