11 – Characterisation using imaging techniques
S. Anderson
2007
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Key Takeaway Key Takeaway: This chapter outlines the importance of adapting microscopy techniques and experimental design for biomaterial samples and tissue engineering constructs, emphasizing the use of Confocal Laser Scanning Microscopy (CLSM) for live cell imaging.
Abstract
Publisher Summary The field of tissue engineering delivers a distinct set of samples to the microscopist. These comprise both biomaterial samples (polymers, metals, and ceramics) in a two- or three-dimensional structure, with or without the addition of biological material (cells and extracellular matrix components) around and within the material scaffold. This composite structure is challenging as, although microscopy is used as a tool to examine both biological and material samples independently, the typical protocols utilized are generally different. For example, the chemicals used to process tissue for histology or electron microscopy may dissolve some polymers or the force required to section engineered scaffolds may shred the delicate cell-matrix layers attached to their surfaces. It is, therefore, necessary to adapt techniques and alter experimental design to address the unique challenge these samples pose. This chapter presents a practical approach to microscopy of biomaterial samples and tissue engineering constructs. A key issue, crucial to the efficiency and success of a cell experiment in tissue engineering is experimental design and the chapter describes the general considerations so that the appropriate technique can be chosen on the basis of what is optimal for the samples and also what is available in the imaging laboratory. A core topic discussed in the chapter is the use of Confocal Laser Scanning Microscopy (CLSM) to image cells in 3D scaffolds and the different options available within this technique. The chapter presents a guide to live cell imaging using vital dyes and on combining CLSM with other techniques to maximize the amount of information gained from a given experiment.