Abstract
Elucidation of intracellular cholesterol transport is important for understanding the molecular basis of several metabolic and neuronal diseases, like atheroclerosis or lysosomal storage disorders. Progress in this field depends crucially on the development of new technical approaches to follow the cellular movement of this essential lipid molecule. In this article, a survey of the various methods being used for analysis of sterol trafficking is given. Various classical biochemical methods are presented and their suitability for analysis of sterol trafficking is assessed. Special emphasis is on recent developments in imaging technology to follow the intracellular fate of intrinsically fluorescent sterols as faithful cholesterol markers. In particular, UV-sensitive wide field and multiphoton microscopy of the sterol dehydroergosterol, DHE, is explained and new methods of quantitative image analysis like pixel-wise bleach rate fitting and multiphoton image correlation spectroscopy are introduced. Several applications of the new technology including observation of vectorial sterol trafficking in polarized human hepatoma cells for investigation of reverse cholesterol transport are presented.
Keywords: Fluorescent sterols, cholesterol ergostero, wide field microscopy, multiphoton microscopy, image correlation spectroscopy, particle tracking, hepatocytes, bile, atherosclerosis, macrophages, vectorial sterol trafficking, sterol dehydroergosterol, lysosomal storage disorders, heavy atomic isotopes, lipid molecule
Current Pharmaceutical Biotechnology
Title: Following Intracellular Cholesterol Transport by Linear and Non-Linear Optical Microscopy of Intrinsically Fluorescent Sterols
Volume: 13 Issue: 2
Author(s): Daniel Wustner
Affiliation:
Keywords: Fluorescent sterols, cholesterol ergostero, wide field microscopy, multiphoton microscopy, image correlation spectroscopy, particle tracking, hepatocytes, bile, atherosclerosis, macrophages, vectorial sterol trafficking, sterol dehydroergosterol, lysosomal storage disorders, heavy atomic isotopes, lipid molecule
Abstract: Elucidation of intracellular cholesterol transport is important for understanding the molecular basis of several metabolic and neuronal diseases, like atheroclerosis or lysosomal storage disorders. Progress in this field depends crucially on the development of new technical approaches to follow the cellular movement of this essential lipid molecule. In this article, a survey of the various methods being used for analysis of sterol trafficking is given. Various classical biochemical methods are presented and their suitability for analysis of sterol trafficking is assessed. Special emphasis is on recent developments in imaging technology to follow the intracellular fate of intrinsically fluorescent sterols as faithful cholesterol markers. In particular, UV-sensitive wide field and multiphoton microscopy of the sterol dehydroergosterol, DHE, is explained and new methods of quantitative image analysis like pixel-wise bleach rate fitting and multiphoton image correlation spectroscopy are introduced. Several applications of the new technology including observation of vectorial sterol trafficking in polarized human hepatoma cells for investigation of reverse cholesterol transport are presented.
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Cite this article as:
Wustner Daniel, Following Intracellular Cholesterol Transport by Linear and Non-Linear Optical Microscopy of Intrinsically Fluorescent Sterols, Current Pharmaceutical Biotechnology 2012; 13 (2) . https://dx.doi.org/10.2174/138920112799095301
DOI https://dx.doi.org/10.2174/138920112799095301 |
Print ISSN 1389-2010 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-4316 |
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