Dojindo Laboratories, Kumamoto, Japan.
Department of Chemistry, Faculty of Science, Fukuoka University, Japan.
FEBS Lett. 2018 Feb;592(4):559-567. doi: 10.1002/1873-3468.12979. Epub 2018 Feb 2.
We have developed two types of fluorescent probes, DALGreen and DAPGreen, for monitoring autophagy, that exhibit fluorescence upon being incorporated into autophagosomes. DALGreen enhances its fluorescence at acidic pH, which is favorable for monitoring late-phase autophagy, whereas DAPGreen remains fluorescent with almost constant brightness during the autophagic process. With these probes that stain autophagosomes as they are being formed, the real-time change of autophagic phenomena of live cells may be traced, which is an advantage over conventional approaches with small molecules that stain mature autophagosomes. The use of both dyes allows monitoring of the membrane dynamics of autophagy in any type of cell without the need for genetic engineering, and therefore, will be useful as a tool to study autophagic phenomena.
我们开发了两种荧光探针，DALGreen 和 DAPGreen，用于监测自噬，它们在被纳入自噬体后会发出荧光。DALGreen 在酸性 pH 值下增强其荧光，这有利于监测晚期自噬，而 DAPGreen 在自噬过程中保持几乎恒定的亮度。使用这些探针可以在自噬体形成时对其进行染色，从而可以实时跟踪活细胞中自噬现象的变化，这是优于传统小分子方法的优势，因为传统方法只能对成熟的自噬体进行染色。这两种染料的使用允许在不需要基因工程的情况下监测任何类型细胞的自噬膜动力学，因此，它将作为一种研究自噬现象的工具很有用。
