Rasya Jauhar
Dissection. The targeted tissues must be dissected and their weights be measured. It could be wet weight or dry weight, it is up to your research. Digestion. The digestion process is required to remove all of the organic materials from the tissues, leaving only the microplastics. It could be done using a wide range of reagents and exposure time to process the sample. Hydrogen peroxide and KOH are the common reagents to use. Density separation (optional). Some of the sample solutions might have organic matters that still cannot be fully digested. You could use NaCl solution to separate the sample's density. Separate it using a separatory funnel and discard the bottom part of the solution; use the supernatant as microplastics tend to float on the solution. Filtration. At this stage, you need filters/membranes with the help of a vacuum pump to filter the digested solution. The three crucial factors for choosing the filters are the pore size, filter material, and filter size. The current ideal filters for microplastic study are Anodisc and silicone filters as they emit low to no interference for FTIR and Raman analysis. Visual Sorting. Data all of the microplastics from the tissue samples using a microscope. Categorize the microplastics by their type, size, and color. Raman Analysis. Not all microplastics found in the tissues must be subjected to Raman analysis and you can sort the microplastics based on your research objective. However, it should be regarded as crucial for pieces smaller than 100 µm, and a percentage (5–10%) of all samples should be routinely evaluated to validate the relative accuracy of any visual assessment. By comparing the unknown sample's spectra to a known reference polymer in a database, microplastics can be identified using Raman spectroscopy.
There are many tissues sample that can be analyzed for microplastic study; each requires its own procedure for it to be prepared. I will answer this with the context of fish tissues. The process includes:
Disclaimer: This answer is based on my undergraduate thesis. If there is any correction or comment feel free to do it as I'm also still learning. Thank you
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