BSA@AMI is a simple protocol that allows for accurate determination of size distributions of nanoparticles. This tool guides you step-by-step through the process of preparing your sample for transmission electron microscopy (TEM) to get an image that can be analyzed manually of with an image analysis software like Fiji or ImageJ
All you need is your sample:
If you are interested in the scientific details, you can read our paper.
In order to get started, you need a first estimate of the particle size (diameter in nm) and the mass density (in g/cm3) of your material, e.g. by scattering.
In the next step, you will calculate the concentration and prepare your BSA solution.
Using the information you gathered about your sample so far, it's now possible to determine the needed concentration of the bovine serum albumin (BSA) solution. Please provide all the input below:
Now you can prepare your sample for microscopy. Just mix your nanoparticle dispersion with the prepared BSA solution in a 1:1 ratio and allow the dispersion to incubate for at least two hoursbefore preparing the TEM grid. In case of incubation overnight, incubate the dispersion in the fridge. You will need 5 µl for the TEM grid.
Analysis of the TEM image can be done manually by counting or in an automated fashion.
There are many different ways to do the image analysis automatically. One very simple way is to use Fiji, apply a binarization to your image via a threshold and run the particle analysis tool. Please visit the Fiji documentation for more details.
If you have any questions or remarks, please get in touch with us. If you used and liked the tool, please cite us.