A novel experimental technique for cell autophagy research

Researchers at the National Institutes of Health and Oxford University in the United Kingdom have developed a high-throughput technology that can quantitatively analyze autophagy in primary human cells. This technology is the first to detect the level of autophagy in diseases known to be related to autophagy, which will help to further analyze cellular autophagy that is closely related to the efficacy of drugs. The relevant results are published in the journal Autophagy.

Cellular autophagy is an evolutionarily conservative process, which includes wrapping the cytoplasmic components in a bilayer membrane structure called autophagosomes, and transporting them to the lysosomes for degradation. Autophagy can regulate cell homeostasis by degrading long-lived proteins, protein polymers, and damaged organelles. It can also inhibit tumor formation by limiting inflammation, removing toxic unfolded proteins, and removing damaged mitochondria that generate reactive oxygen species (which can damage DNA), so losing these protective measures will promote cancer.

But the traditional cell autophagy detection technology is not suitable for clinical samples, which also prevents the further development of cell autophagy research. In this article, the researchers used Amnis's (now Merck Millipore) ImageStream imaging analysis flow cytometer to propose a high-throughput, quantitative analysis of primary human cell autophagy.

The researchers have verified in multiple cell lines knocked out of the basic cell autophagy gene, as well as primary cells, showing that this method can effectively provide statistical data of cell populations, and can also obtain images of individual cells, thereby providing cell autonomy. Complete information.

In this way, the researchers found that T cells have stronger autophagy activity than B cells, and the main senescent cells CD8 + T cells in healthy samples have decreased autophagy activity as DNA damage increases, which may explain the aging immune system. Some of the relevant characteristics of the system, and why this system ages with age.

The researchers believe that this technology has detected the level of autophagy for the first time in diseases known to be related to cell autophagy, which will help to further analyze the close relationship between cell autophagy and drug efficacy.

ImageStream is a brand-new loss-of-cell technology that combines flow cytometry with fluorescence microscopy to provide statistical data on cell populations and obtain images of individual cells to provide cell morphology , Cell structure and subcellular signal distribution information.

At present, ImageStream has launched the latest version: ImageStreamX Mark II, which is the third generation ImageStream imaging flow cytometer, with unique cell analysis capabilities. Compared with the original ImageStream, Mark II produces multiple high-resolution images of each flowing cell, including bright field, dark field, and up to 10 fluorescent markers, and the sensitivity is more than traditional flow cytometry. Compared with ImageStreamX, Mark II's process is simpler, more flexible, and optimized for rare cell applications. ImageStreamX Mark II can capture up to 12 high-resolution images of each flowing cell in real time, with a detection rate of up to 5000 cells / second and enhanced fluorescence sensitivity. These functions of ImageStreamX Mark II can detect cell morphology, intensity and location of fluorescent probes, and thus provide scientists with a wide range of image analysis applications, including cell-cell interactions, phagocytosis, apoptosis and autophagy, nuclear translocation, morphology Change etc.

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