Quantitative Analysis of Hypothalamic Neuropeptides

Slides were anatomically-matched and analyzed by an investigator blinded to the experimental groups. Sections were imaged using a Zeiss Axio Imager M2 microscope connected to an ORCA-R2 CCD digital camera (Hamamatsu Photonics). Gray-scale images of cell bodies, when present, and fibers immunopositive for AgRP, MSH, and AVP (Supplemental Fig. 6) were analyzed in Image J 1.43 u (National Institutes of Health, Bethesda, MD). Iba-immunoreactivity was analyzed as in^{43 (link)}. The region of analysis was outlined in each section. Subjects for which the relevant sections were damaged or unavailable were dropped from a given analysis. All immunoreactivity is quantified by the mean area stained in pixels.

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Holder M.K., Peters N.V., Whylings J., Fields C.T., Gewirtz A.T., Chassaing B, & de Vries G.J. (2019). Dietary emulsifiers consumption alters anxiety-like and social-related behaviors in mice in a sex-dependent manner. Scientific Reports, 9, 172.

Publication 2019

Axio Imager M2 microscope

Agrp Bodies images Camera Cell Microscope Orca

Corresponding Organization :

Other organizations : Georgia State University

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Variable analysis

independent variables

Experimental groups

dependent variables

Quantification of immunoreactivity for AgRP, MSH, and AVP
Quantification of Iba-immunoreactivity

control variables

Anatomically-matched slides
Investigator blinded to the experimental groups
Imaging using a Zeiss Axio Imager M2 microscope and ORCA-R2 CCD digital camera
Image analysis using ImageJ 1.43u

controls

Positive control: Iba-immunoreactivity analyzed as in the referenced study (43)
Negative control: Not explicitly mentioned

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