Automated axon length quantification for populations of labelled neurons

Broser, P. J., Erdogan, S., Grinevich, V., Osten, P., Sakmann, B., Wallace, D. J. (March 2008) Automated axon length quantification for populations of labelled neurons. Journal of Neuroscience Methods, 169 (1). pp. 43-54. ISSN 0165-0270

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Abstract

Virus-based methods for labelling populations of cortical neurons, when combined with cell-type specific recombinant promoters and techniques allowing temporal control of gene expression, provide neuroscience with new opportunities to examine the connectivity between brain regions and how this connectivity is modified by experience or disease. However, to take full advantage of these technical advances, it is necessary to develop new methods for quantification of the axonal projections revealed. Here we describe a method for quantitative analysis of axonal projection patterns emanating from populations of labelled cells, using transmitted light bright field microscopy. A single high resolution image of an area to be analysed is first acquired using mosaic extended focus image microscopy. This image is then analysed by specifically developed image processing algorithms that identify and track axon segments present. For quantitative analysis, measurement grids consisting of a user-defined number of individual elements are placed over an area of interest, with the computer-based method then returning the summed length of the axon segments in each element. Axon density plots can thus be generated. We present an example from rat brain showing, over a whole coronal section, axon densities emanating from a population of layer 2/3 somatosensory neurons.

Item Type:	Paper
Uncontrolled Keywords:	Algorithms Animals Animals, Newborn Axons/physiology/*ultrastructure Brain Mapping/instrumentation/*methods Cell Count/instrumentation/*methods Cell Shape/physiology Cell Size Genetic Vectors/genetics Green Fluorescent Proteins/genetics Image Cytometry/instrumentation/*methods Image Processing, Computer-Assisted Lentivirus/genetics Microscopy/instrumentation/*methods Pyramidal Cells/physiology Rats Rats, Wistar Somatosensory Cortex/cytology/physiology Staining and Labeling/instrumentation/*methods
Subjects:	bioinformatics > genomics and proteomics > annotation > gene expression profiling annotation Investigative techniques and equipment > microscopy organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > kinase > tyrosine kinase organism description > virus
CSHL Authors:	Osten, Pavel
Communities:	CSHL labs > Osten lab
Depositing User:	Tom Adams
Date:	30 March 2008
Date Deposited:	26 Aug 2011 13:31
Last Modified:	05 Apr 2013 16:57
Related URLs:	Publisher
URI:	https://repository.cshl.edu/id/eprint/7694

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