can we measure vascular smooth muscle cells proliferation with haematoxylin eosin staining? i want to measure vascular muscle cells proliferation with simple HE staining, can I? or other staining can measure?
Are there any methods for know the different hyperplasia and hypertrophy in cross section in artery?
This is a difficult question, and it is not entirely clear what your starting hypothesis is. If what you want to know is whether there is generalized hypertrophy/hyperplasia that is causing the wall to thicken, then you can simply measure the thickness of the wall, using basic morphometry tools. Any standard histologic stain would work for this, but you have to make sure that the arteries are cut at exactly the same site from animal to animal, that the sections are perfectly transverse, and that the arteries being compared have similar lumen diameters. The lumen diameter is particularly important; in any normal animal, the artery diameter will fluctuate; if an artery has been sampled and fixed in a dilated state, with its lumen wider than that of another animal (just from normal individual physiologic variability), its wall will be more stretched and thinner than the other artery, even if both arteries are normal. Ideally you may want to normalize the wall thickness measurement to the lumen diameter.
If what you are interested is at the cellular level (presence of proliferation), then you can stain by immunohistochemistry with a proliferation marker (ki67, PCNA) and count the number of positive nuclei over the total number of nuclei. Since in one section of artery you won't have that many nuclei, you would need to do a number of sections to attain a reasonably representative number of nuclei. Evaluting for hypertrophy at the cellular level would be even more difficult.
I guess this will be difficult. If you want to rely on H&E stained sections, you will have to count mitotic figures. I would recommend to think about using immunohistology with a Ki-67 antibody. To be more specific, I would need to know a little bit more about what question you want to address and what model you are studying.
Given the purpose of the H/E staining, I doubt you can be able to quantify or distinguish such a cell type with a generic staining like this.
Mr Geral Niedobitek:i want to know what happen with the VSMC in artery when we know the tunica media are more thickness than control?can i have your email address? thanks for your answer sir
This is a difficult question, and it is not entirely clear what your starting hypothesis is. If what you want to know is whether there is generalized hypertrophy/hyperplasia that is causing the wall to thicken, then you can simply measure the thickness of the wall, using basic morphometry tools. Any standard histologic stain would work for this, but you have to make sure that the arteries are cut at exactly the same site from animal to animal, that the sections are perfectly transverse, and that the arteries being compared have similar lumen diameters. The lumen diameter is particularly important; in any normal animal, the artery diameter will fluctuate; if an artery has been sampled and fixed in a dilated state, with its lumen wider than that of another animal (just from normal individual physiologic variability), its wall will be more stretched and thinner than the other artery, even if both arteries are normal. Ideally you may want to normalize the wall thickness measurement to the lumen diameter.
If what you are interested is at the cellular level (presence of proliferation), then you can stain by immunohistochemistry with a proliferation marker (ki67, PCNA) and count the number of positive nuclei over the total number of nuclei. Since in one section of artery you won't have that many nuclei, you would need to do a number of sections to attain a reasonably representative number of nuclei. Evaluting for hypertrophy at the cellular level would be even more difficult.
Proliferation will not be accessible by using a standard stain like HE.
Counting of mitotic figures will be an issue, due to the relatively slow proliferation rate in the smooth muscles
As suggested in previous answers assessment of proliferation markers is the way to go.
Other is to look for a vascular remodeling, manifested by thickened tunica media and, eventually, by displacement of smooth muscle cells into the intima - intimal outgrowth of smooth muscles cells.
PAS staining will give you the borders of the smooth muscle cells in the media of the vessel. Simple counting of the cells in several (ate least 3) serial sections will give you some idea, whether you have a productive (hyperplastic) or non-productive (hypertrophic) vascular alteration.
If there is no possibility for PAS stain, then just count the nuclei in HE, which will give you a number of smooth muscle cells. However, I will prefer PAS, since one can really see, whether the cells are hypertrophic ot not , just by delineation of their cell borders.
I hope this might help you
Best and good luck!
H&E stain allows to identify smooth muscle cells. You may measure their area by point counting or image analysis in the vessels of same caliber, in horizontal sections. The difference of area will be the reflection of the increase of sooth muscle amount.
Because i think tunica media increase size it can be hypertrophy or hyperplasia, or proliferation, i can not identify clear boundary cell with HE
Hi,
Why not try these if you can afford the antibodies.
http://www.ncbi.nlm.nih.gov/pubmed/23749886
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1360224/
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