How can I calculate the aspect ratio of silver nanoparticles internalized in Hela cell lines?? I have functionalized the silver nanoparticles with different poly aminoacids. Now I want to calculate aspect ratio of particles internalized through TEM images of HeLa cells.
Within ISO 9276-6 the definition of aspect ratio is breadth/length so that all answers lie between 0 and 1 for shape parameters. Aspect ratio is a 2-D property of 3-D particles. You can do this manually by counting enough particles (10000 images will give you a standard error of 1% on the mean) or automatically perhaps with a program like Image J (NIST freeware). See for example:
Jun 14th, 2007 Shape aspects of particle size analysis - the math http://www.brainshark.com/brainshark/vu/view.asp?pi=370539738&text=M021507
November 19th, 2015 The mathematics of image analysis
https://www.brainshark.com/malvern/vu?pi=312905118&b=1&tx=120486&c1=22
There are many image analysis programs available these days that will quite readily give you the information you need - usually at least one from each manufacturer of every decent microscope, and many more besides those. Some are free but they require a little more effort to fully understand and use them.
You should calculate aspect ration of nanoparticles before in vitro or in vivo cell permeations studies. You can use TEM or AFM for imaging and calculate aspect ration as advised by Alan F Rawle and Tausif Alam. However, if you believe that nanoparticls with slight differences in aspect ratio show different permeation, you will need to break/digest/homogenize the cells or tissue, In this way, you will get nanoparticles that were inernalized and proceed to further imaging and calculation of aspect ratio.
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