Real-time calcium measurements of live optically trapped microorganisms.

摘要: A system has been developed that allows for the real-time measurement of calcium dynamics in swimming sperm. Specifically, ratiometric dye Indo-I is used as a fluorescent indicator intracellular dynamics. The dual emissions are collected by high-sensitivity back-illuminated CCD camera coupled to Dual-View imaging system. From CCD, images sent custom algorithm which processes and outputs measurements real-time. Additionally, sperm velocity position data processed outputted obtained using separate red light (>670 nm) phase contrast setup does not optically interfere with imaging. Using this effects optical trapping on was determined. Optical decaying focused laser power 510 mW 3 over 8 seconds causes statistically insignificant change between in-trap out-of-trap conditions. Progesterone, activator, added were trapped under second decay Progesterone treated higher average level than untreated sperm, but shows no statistical difference progesterone Trapping at 16 without decay, have shown decrease motility, baseline pre-trap levels.