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Light microscopic image analysis system to quantify immunoreactive terminal area apposed to nerve cellsThe present report describes a desktop computer-based method for the quantitative assessment of the area occupied by immunoreactive terminals in close apposition to nerve cells in relation to the perimeter of the cell soma. This method is based on Fast Fourier Transform (FFT) routines incorporated in NIH-Image public domain software. Pyramidal cells of layer V of the somatosensory cortex outlined by GABA immunolabeled terminals were chosen for our analysis. A Leitz Diaplan light microscope was employed for the visualization of the sections. A Sierra Scientific Model 4030 CCD camera was used to capture the images into a Macintosh Centris 650 computer. After preprocessing, filtering was performed on the power spectrum in the frequency domain produced by the FFT operation. An inverse FFT with filter procedure was employed to restore the images to the spatial domain. Pasting of the original image to the transformed one using a Boolean logic operation called 'AND'ing produced an image with the terminals enhanced. This procedure allowed the creation of a binary image using a well-defined threshold of 128. Thus, the terminal area appears in black against a white background. This methodology provides an objective means of measurement of area by counting the total number of pixels occupied by immunoreactive terminals in light microscopic sections in which the difficulties of labeling intensity, size, shape and numerical density of terminals are avoided.
Document ID
20040172947
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
Authors
Wu, L. C.
(San Jose State University Foundation CA 95192, United States)
D'Amelio, F.
Fox, R. A.
Polyakov, I.
Daunton, N. G.
Date Acquired
August 22, 2013
Publication Date
June 6, 1997
Publication Information
Publication: Journal of neuroscience methods
Volume: 74
Issue: 1
ISSN: 0165-0270
Subject Category
Life Sciences (General)
Distribution Limits
Public
Copyright
Other
Keywords
Non-NASA Center
NASA Discipline Neuroscience
NASA Center ARC

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