Review
Advanced microscopy techniques for quantitative analysis in neuromorphology and neuropathology research: current status and requirements for the future

https://doi.org/10.1016/j.jchemneu.2010.06.005Get rights and content

Abstract

Visualizing neuromorphology and in particular neuropathology has been the focus of many researchers in the quest to solve the numerous questions that are still remaining related to several neurological and neuropsychiatric diseases. Over the last years, intense research into microscopy techniques has resulted in the development of various new types of microscopes, software imaging systems, and analysis programs. This review briefly discusses some key techniques, such as confocal stereology and automated neuron tracing and reconstruction, and their applications in neuroscience research. Special emphasis is placed on needs for further developments, such as the demand for higher-throughput analyses in quantitative neuromorphology. These developments will advance basic neuroscience research as well as pharmaceutical and biotechnology research and development.

Introduction

In neuromorphology and neuropathology, quantitative analyses are the primary factor towards advanced understanding of normal and pathological composition of neural tissue. Design-based stereology has become widely accepted in analyzing populations of three-dimensional (3D) structures such as neurons, while neuron tracing and reconstruction are utilized for morphological analyses of single neurons or networks of neurons. Many new insights into disease pathogenesis were gained through quantification of profound or, in particular, subtle changes in regional volumes of brain regions, numbers of neurons and glial cells, size of cells, neuron arborization, spine numbers, etc. Although new techniques have evolved during recent years, efficiency and quality of these techniques can still be improved.

In this review we first briefly explain the meaning of design-based stereology, neuron tracing and reconstruction. Subsequently we focus on past, present, and possible future developments in the aforementioned disciplines. Moreover, we give examples how researchers apply these techniques in neuroscience, and finally outline some needs for the future in order to perform higher-throughput analyses.

Section snippets

Design-based stereology

Design-based stereology is a discipline to perform precise quantitative evaluations of the structure of small and large 3D objects, hence the name stereology (stereos = solid; logos = knowledge) (Weibel, 1979, Howard, 1998, Schmitz and Hof, 2005, Glaser et al., 2007).

By analyzing systematically and randomly sampled two-dimensional (2D) sections or 3D subsamples of tissue, one can calculate morphometric parameters of a 3D structure, while eliminating almost all potential methodological bias. In

Neuron tracing and reconstruction

Neuron tracing and reconstruction is the process of delineating and reconstructing the cell body of a neuron, its axon, dendrites and spines, hereby creating a geometric model (Glaser and Glaser, 1990). It has become increasingly apparent that the morphological properties of a single cell determine the neuronal dynamics and synaptic plasticity of this particular cell (see, e.g., Mainen and Sejnowski, 1996, Koch and Segev, 2000, Euler and Denk, 2001, Vetter et al., 2001, Krichmar et al., 2002,

Conflict of interest

C.S. serves as paid consultant for MBF Bioscience (Williston, VT, USA), some products of which are discussed in this review. However, no financial support has been received directly or indirectly related to this manuscript.

Acknowledgements

The illustration was made by an SI-SD system (MBF Bioscience), which was obtained by an NWO grant no. 91106003 “Semiautomated Confocal Microscopical High-Precision Design-Based (CM-HPDB) Stereology – applications in neuroscience with a focus on neurodegenerative diseases”.

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