Tracking metabolic dynamics of apoptosis with high-speed two-photon fluorescence lifetime imaging microscopy.

摘要: Programmed cell death, or apoptosis, is an essential process in development and homeostasis, disruptions associated pathways are responsible for a wide variety of diseases such as cancer, developmental abnormalities, Alzheimer’s disease. On the other hand, many cases, desired outcome therapeutic treatments targeting cancer. Recently, metabolic imaging based on two-photon fluorescence microscopy has been developed shown to be highly sensitive certain death processes, most notably thus having potential advanced label-free screening tool. However, typically low acquisition rates this technique have resulted limited throughput approach, allowing only small population cells tracked at well-separated time points. To address limitation, high-speed lifetime (2P-FLIM) platform capable video-rate applied study further characterize dynamics with death. Building upon previous work demonstrating capabilities system, microscope utilized rapid changes during induction, dose-dependency response, response invasive vs. noninvasive cancer cells, apoptosis-resistant line, which undergo autophagy toxic stimuli. Results from these experiments show that early apoptosis-related strongly correlated important cellular parameters including responsiveness apoptosis-inducing The high speed sensitivity presented approach enables new investigations into dynamic complex process.