Automated Quantification of Glial Activation and Storage Material

To analyze the degree of glial activation in the gray matter (GFAP-positive astrocytes + CD68-positive microglia) as well as accumulation of storage material (SCMAS), a semiautomated thresholding image analysis method was used with Image-Pro Premier software (Media Cybernetics)^{23 (link)}. Briefly, this involved the collection of slide-scanned images at 10 × magnification for a one-in-six series of sections per animal followed by demarcation of all regions of interest⁴⁷ while maintaining the lamp intensity, video camera setup, and calibration constant throughout image capturing. Images were subsequently analyzed using Image-Pro Premier (Media Cybernetics) using an appropriate threshold that selected the foreground immunoreactivity above background. This threshold was then applied as a constant to all subsequent images analyzed per batch of animals and reagent used to determine the specific area of immunoreactivity for each antigen^{23 (link)}. Specific regions of interest included the ventral posteromedial/posterolateral (VPM/VPL) nuclei of the thalamus and the S1BF cortex^{22 (link),31 (link),32 (link)}.

Free full text: Click here

Dearborn J.T., Nelvagal H.R., Rensing N.R., Takahashi K., Hughes S.M., Wishart T.M., Cooper J.D., Wong M, & Sands M.S. (2022). Effects of chronic cannabidiol in a mouse model of naturally occurring neuroinflammation, neurodegeneration, and spontaneous seizures. Scientific Reports, 12, 11286.

Publication 2022

Image-Pro Premier

Animals Astrocytes Gfap Glial Gray matter Microglia Nuclei of thalamus

Corresponding Organization : Washington University in St. Louis

Other organizations : University of Otago, Roslin Institute, University of Edinburgh

Top 5 similar protocols

Variable analysis

independent variables

Degree of glial activation in the gray matter (GFAP-positive astrocytes + CD68-positive microglia)
Accumulation of storage material (SCMAS)

dependent variables

Area of immunoreactivity for each antigen (GFAP, CD68, SCMAS)

control variables

Lamp intensity
Video camera setup
Calibration constant throughout image capturing
Threshold applied as a constant to all subsequent images analyzed per batch of animals and reagent used

controls

No positive or negative controls were explicitly mentioned.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!