Hi everyone,
1.I am doing spectral imaging and linear unmixing on overlapped images and I wonder if possible to quantify the contribution of each component.
I explain. I am trying to separate elastin from other sources of fluorescence and then quantify elastin as pixel over the total amount of pixel in the image, then calculate the percentage.
Am I making any sense?
Does anyone know any specific technique to do this?
2.Also, I would like to measure the collagen crimp on SHG images but I am not sure how to do it. I found lots of papers about this but my images are not zoomed enough to actually see collagen bands and I was wondering if anyone knows a way to do it.
Thank you
Hi, I think your samples should be studied by video image analysis. I recommend that you use OrientationJ, which is a Java plug-in for ImageJ software. I'm using OrientationJ to evaluate collagen fibers in polarized light microscopy and SHG. The method is pixel-wise. The software computes the structure tensor (a 2x2 matrix) for each pixel in the image by sliding the Gaussian analysis window over the entire image and provides data on coherence (isotropy / anisotropy), energy (structure tensor trace), and local orientation. Through the coherence analysis you will be able to observe information about the crimp of the collagen fibers. Through Energy maps you will be able to discriminate the pixels associated with elastin.
For details I recommend that you read:
http://bigwww.epfl.ch/demo/orientation/index.html?image=3
Z. Püspöki, M. Storath, D. Sage, M. Unser, "Transforms and Operators for Directional Bioimage Analysis: A Survey," Advances in Anatomy, Embryology and Cell Biology, vol. 219, Focus on Bio-Image Informatics, Springer International Publishing, ch. 3, pp. 69-93, May 21, 2016.
R. Rezakhaniha, A. Agianniotis, J.T.C. Schrauwen, A. Griffa, D. Sage, C. V.C. Bouten, F.N. van de Vosse, M. Unser, N. Stergiopulos, "Experimental Investigation of Collagen Waviness and Orientation in the Arterial Adventitia Using Confocal Laser Scanning Microscopy," Biomechanics and Modeling in Mechanobiology, Vol. 11, no. 3-4, pp. 461-473, 2012.
E. Fonck, G.G. Feigl, J. Fasel, D. Sage, M. Unser, D.A. Rüfenacht, N. Stergiopulos, "Effect of Aging on Elastin Functionality in Human Brain Arteries," Stroke, vol. 40, no. 7, pp. 2552-2556, July 2009.
Good luck!
Hi Marcela,
Thanks for thus,it's helpful.
However how can you gain info on elastin through the energy map?
Thank you
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