I am working on primary hippocampus neurons from mice. I am trying to look at the effect of amyloid beta on the neurons. I am looking at parameters like cell death, number of neurons in a field of view, neuronal length. I also want to try to analyze the neuronal coverage or area occupied by neurites to understand how amyloid beta affects the neurons.
Does anyone know how to measure the neuronal coverage or area occupied by neurites?
Thank You.
Hi there,
It's better if you provide an image and explain over the image, more or less, what you want to do in order to get help from the forum.
Cheers,
J
Try tu use the Scholl analysis. Do you wish to know the reference ?
Mihir,
There are a few ways to figure out the area fraction of neuron coverage. The easiest and least costly would be to stain with NueN and use ImageJ to calculate the area fraction of an outlined area such as the hippocampus. Also, Microbrightfield software has Neurolucida which can trace and measure neuron length. Double immunofluorescence staining of A-beta and NueN would show the relationship between Neurons and plaques. UIC here at the UMN has image scanning of slides which can then be used for ImageJ analysis. I have a protocol written to show you how to use this if you want to go that route. Joyce
Hello Mihir,
That's a good question! As suggested by Joyce, you can also dually stain your neurons with GFAP and Beta-amyloid plaques preferably by confocal microscopy as you can have high chances of measuring the spatial distribution of beta-amyloid plaques in 3-D format.
Best
Raghuram
Thank You for the suggestions.
The attached picture is of a neuronal culture. I want to know how to measure the coverage?