Researchers at Japan’s Advanced Institute of Science and Technology have developed a method to isolate intact lysosomes from cells. The technique is fast and produces samples of high purity. Lysosomes are the waste-removing organelles within a cell and are involved in many diseases, from lysosomal storage diseases to autoimmune disorders, certain cancers, and neurodegenerative diseases. However, they are difficult to study, as current techniques for isolating lysosomes from cells result in low-purity samples and damaged or altered lysosomes. This new approach uses nanotechnology to rapidly extract a sample of high-purity lysosomes from cells.
Lysosomes digest large molecules within cells and produce metabolites. When this process goes wrong, a serious illness can occur, which the researchers try to study in detail. However, isolating these small organelles is a challenge. One approach is to centrifuge samples of lysed cells very quickly, which separates lysosomes based on their density. However, this is a fairly crude way of isolating lysosomes, and many other organelles are often mixed in isolation. It’s also pretty slow.
Another technique involves using magnetic beads coated with antibodies that bind to lysosomes, allowing researchers to isolate them with a magnet. This results in a purer isolate, but alters lysosome proteins, affecting any protein analysis that scientists want to perform. To address these issues, researchers in Japan have developed a new method for isolating lysosomes.
They have created hybrid magnetic-plasmonic nanoparticles that contain an alloy of iron-cobalt and silver. The particles are also coated with amino dextran and, unlike other magnetic particles used to isolate lysosomes, the cells are happy to endocytose them, that is, they are ingested and eventually end up in an intact lysosome. The technique then involves breaking the cells and harvesting the intact lysosomes using a magnet.
Particles can also be visualized by plasmonic imaging, allowing researchers to track their progress through a cell until they bind within a lysosome. This image meant that the researchers were able to calculate the best time to lyse the cells and harvest the lysosomes, as they knew exactly when they were inside the lysosomes. The method is very fast and results in very pure lysosome samples.
“We found that the maximum time required to isolate lysosomes after cell rupture was 30 minutes, which is substantially shorter than the time required by centrifugation-based techniques, which typically require a minimum separation time of several hours. said Shinya Maenosono, a researcher. involved in the study. “Given the profound relationship of lysosomes with many cellular metabolites, a deeper understanding of lysosomal function is needed to determine their regulation in different cellular states. Therefore, our technique can contribute to a better understanding and treatment. of lysosomal diseases in the future “.