I want to know, after treatment with 5 groups of rattus novergicus, example:
I want to measure cells proliferation in vascular smooth muscle in carotid artery.
Questions:
1. I know little about histological section with HE to measure proliferation by thickness arterial wall/lumen diameter but i don't know how to measure VSMC in tunica media to measure cells proliferation with HE?are there another staining? and how to interpret the cell proliferation? which software or hardware to analysis?
2. What others assay than can be use to measure proliferation from in vivo example from VSMC in carotid artery?
Thanks for your answers
Hi.
You could use BrdU/EdU-stain. Our model was rat injured carotid. For the portocol have a look on "Local inhibition of ornithine decarboxylase reduces vascular stenosis in a murine model of carotid injury. Forte et al.,2013 Int J.Cardiol".
For the proliferation of VSMC I would suggest you to use EdU which is much better than BrdU for several reason, one of them is that with EdU you do not need to denature DNA which is asked for BrdU. The EdU Assay from Invitrogen is a novel alternative to the BrdU (bromo-deoxyuridine) assay. EdU is a nucleoside analog of thymidine and is incorporated into DNA during active DNA synthesis/cells in proliferation. You could stain also markers of syntheitic/proliferating cells such as PCNA/OPN o matrix proteins.
Hi Ricky,
PAS stain,
Aldehyde Fuchsin-Orcein (quite good for measurement of vascular morphometry)
Productive markers are not so applicable.
Anti-smooth muscle actin antibody can be of help too.
Other suggestion is to contact a veterinary pathologist to help :)
Best regards,
Ivan
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