Mitophagy is the process by which cells remove damaged or aged mitochondria and recycle their constituent elements. It is closely related to many physiological and pathological processes such as aging, neurodegenerative diseases, and cancer, so the study of mitophagy is of great significance.
Currently, two methods are mainly used to study mitophagy:
Antibody or fluorescent protein labeling method
A common detection method using mitochondrial fluorescent probes and lysosomal fluorescent probes
However, both methods have drawbacks. The former is complex to operate and has limited application in living cells; while the latter has low specificity.
The probe molecule HQO developed by Liu Xiangjun and Associate Professor Bing Tao of the Institute of Chemistry, Chinese Academy of Sciences, can accurately locate mitochondria and accurately track the mitophagy process (Figure 1); it provides a new research method for the mitochondrial metabolic process and is also useful for screening mitophagy. Inhibitors or accelerators provide effective tools.
It does not need to be used in combination with other probe molecules. After entering living cells, it is selectively enriched in mitochondria; when mitochondria undergo autophagy to form autophagolysosomes, the pH of the microenvironment decreases, causing HQO to be protonated; the fluorescence of HQO after protonation Both excitation and emission wavelengths are red-shifted (Stokes shift greater than 100 nm).
Due to the different pH values of the microenvironment, HQO displays different colors of fluorescence in mitochondria and autophagy lysosomes (Figure 2), so it can well distinguish between normal mitochondria and autophagy lysosomes that coat damaged mitochondria [1][1][1].
J&K Scientific provides scientists around the world with the fluorescent probe HQO transformed from the scientific research results of Associate Professor Liu Xiangjun:
HQO, 95%
2 MG
Product instructions
Cell culture: Plant the cells in a confocal culture dish, and plant the cells at the corresponding density and culture time according to the needs of different experiments.
Induction of autophagy, cell staining and confocal imaging: induce autophagy under appropriate conditions (such as serum-free medium, PBS or autophagy activator treatment, etc.), add HQO (10 μM) to the serum-free medium, and incubate at 37°C for 10 min, washed 1-3 times with PBS, and confocal imaging was performed. Excitation at 559 nm, emission wavelength of 570-610 nm is the fluorescence that traces normal mitochondria; excitation at 635 nm, emission wavelength of 650-730 nm is the fluorescence that traces autophagy lysosomes.
References
向阳 翟
