Second Harmonic Generation Microscopy for Quantitative Analysis of Collagen Fibrillar Structure

Overview

Journal Nat Protoc

Specialties Biology
Pathology
Science

Date 2012 Mar 10

PMID 22402635

Citations 395

Authors

Xiyi Chen

Oleg Nadiarynkh

Sergey Plotnikov

Paul J Campagnola

Affiliations

Soon will be listed here.

Abstract

Second-harmonic generation (SHG) microscopy has emerged as a powerful modality for imaging fibrillar collagen in a diverse range of tissues. Because of its underlying physical origin, it is highly sensitive to the collagen fibril/fiber structure, and, importantly, to changes that occur in diseases such as cancer, fibrosis and connective tissue disorders. We discuss how SHG can be used to obtain more structural information on the assembly of collagen in tissues than is possible by other microscopy techniques. We first provide an overview of the state of the art and the physical background of SHG microscopy, and then describe the optical modifications that need to be made to a laser-scanning microscope to enable the measurements. Crucial aspects for biomedical applications are the capabilities and limitations of the different experimental configurations. We estimate that the setup and calibration of the SHG instrument from its component parts will require 2-4 weeks, depending on the level of the user's experience.

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