Quantifying Neurochemical Expression Patterns

The computerized image analysis for quantifying the expression pattern of neurochemicals was carried out in those animals subjected to immunohistochemistry (n = 18). The general approach for quantitative image analysis was similar to our previous studies [68 (link),71 (link)]. A computer based image analysis system along with the Image-Pro Plus software 6.0 (Media Cybernetics, Silver Spring, MD, USA) was used to quantify the amount of positive labeling. A digital camera mounted on a ZEISS microscope (Axioplane 2, Carl Zeiss MicroImaging GmbH, Hamburg, Germany) imaged sections and displayed them on a high-resolution monitor. A total of 20 sections per animal, representing the entire rostro-caudal extent of the CeA, were analyzed. The number of CeA neurons (or nuclei) reacted for CRF, SP, and c-fos was then densitometrically counted, and all readings were combined and averaged to obtain the total amount of labeling. For quantifying the morphological changes of the microglia (ranging from simple rounded to complex branched), the value of fractal dimension (D) was used as an indicator in which a higher D means a greater complexity of the expression pattern [72 (link)]. All images were captured on the same day by the same experimenter to maintain the uniform settings adjusted at the beginning of capturing.

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Liao W.C., Yao R.A., Chen L.Y., Renn T.Y., Klimenkov I.V., Sudakov N.P., Mai F.D., Chen Y.T, & Chang H.M. (2022). Olfactory Stimulation Successfully Modulates the Neurochemical, Biochemical and Behavioral Phenotypes of the Visceral Pain. Molecules, 27(21), 7659.

Publication 2022

Image-Pro Plus software 6.0 Axioplane 2

Animal C fos Camera Immunohistochemistry Microglia Microscope Neurons Nuclei Silver

Corresponding Organization : National Taipei University of Nursing and Health Science

Other organizations : National Chung Hsing University, Chung Shan Medical University, Hiroshima University, Siberian Branch of the Russian Academy of Sciences, Limnological Institute

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Variable analysis

independent variables

Immunohistochemistry

dependent variables

Expression pattern of neurochemicals
Morphological changes of the microglia (ranging from simple rounded to complex branched)
Number of CeA neurons (or nuclei) reacted for CRF, SP, and c-fos

control variables

Uniform settings adjusted at the beginning of capturing images
All images were captured on the same day by the same experimenter

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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