Photostimulation by femtosecond laser triggers restorable fragmentation in single mitochondrion

Mitochondrial research is important to the study of ageing, apoptosis, and metabolic diseases. Over the years, mitochondria have been studied with stimulation by chemical agents in a global manner for basic and applied research. This approach lacks of precision and accuracy in terms of spatial and temporal resolution. Here we demonstrate a direct and well-defined photostimulation targeting on single mitochondrial tubular structure using a tightly-focused femtosecond (fs) laser that could precisely activate mitochondria at single tubule level to show restorable fragmentation and subsequent recovery after tens of seconds. In these two processes, a series of mitochondrial reactive oxygen species (mROS) flashes was observed and found critical to the mitochondrial fragmentation. Meanwhile, transient openings of mitochondrial permeability transition pores (mPTP) were seen with oscillations of mitochondrial membrane potential. These activities were crucial for the recovery through scavenging the mROS. Without the feedback mechanisms, the fragmented mitochondria could not return back to their original tubular structure. These interesting observations show that photostimulation by fs laser is an active, precise, clean and well-defined approach to dissect the role of mitochondria in normal physiology and different kinds of diseases.