Hi, I want to quantify total tau, phospho tau and tubulin in the axon and dendrite of hippocampal and cortical neurons by Confocal microscopy analysis. I'll appreciate if you could suggest me the method to distinguish axon from dendrites, and the method to quantify them using imageJ software.
I have attached the image file/picture I scanned using microscope (red channel: total tau, green channel: tubulin, yellow channel: phospho tau). The image is focused on neurites.
Thank you.
Sincerely,
Saroj
There are microfluidic chambers that allowes the separation of cell body and dendrites from axons.
Sigma also has provided an assay kit for this purpose
https://www.sigmaaldrich.com/technical-documents/articles/biology/cell-culture/cellular-assays/neurite-outgrowth-assays.html
The picture you have shown looks like a picture of basket cells. Do you want to differentiate Axons and dendrites in these cells only?
I agree with what Dr. Mona says.
Do you have individual channel images? One with red only, one with green only, and one with yellow only? These are used for quantification. Otherwise, you will need to use the Wand Tracing option in ImageJ.
https://imagej.nih.gov/ij/docs/guide/146-19.html
Hi Nurul, yes I have individual channel for green green and yellow. Just wondering how to separate axon from dendrites and other neurites...
Hi Mona and Muneeb. Thank you so much for your suggestion.
I was wondering if the filaments with microfluids chambers are axon, I could differentiates axon. I took the thick filaments which are at proximity of soma (cell body) as dendrites. So I am confused in the selection appropriate axons. In the image I have attached, soma in not under focus as I scanned the image with prim focus on neurites (axon and dendrites). I have already taken and scanned images for only cell body(soma). That's why I need to distinguish axon in the image I have attached. Your suggestion would be highly appreciable. Thank you once again.
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