RESUMO
The development of a 1.65 µm cavity ringdown methane spectrometer for methane isotope analysis is reported. In order to reduce the laser linewidth, simple optical feedback with an 11 m external fiber cavity using a retroreflector was implemented and it improved the sensitivity. The detection limit at the ppt level for both 12CH4 and 13CH4 concentrations at 100 Torr gas pressure was evaluated from the Allan-Werle plot calculated from the dataset obtained at the fixed laser frequency. In contrast, the detection limit estimated from the baseline noise on the absorption spectrum was a few ppb for both methane isotopologues due to the periodic background oscillations that remained even after baseline correction. The system demonstrated the direct measurement of ambient methane in atmospheric room air, and the estimated 13CH4 ratio as well as the methane concentration were in good agreement with the reference values of ambient air.
RESUMO
Intracellular signaling plays essential roles in various cell types. In the central nervous system, signaling cascades are strictly regulated in a spatiotemporally specific manner to govern brain function; for example, presynaptic cyclic adenosine monophosphate (cAMP) can enhance the probability of neurotransmitter release. In the last decade, channelrhodopsin-2 has been engineered for subcellular targeting using localization tags, but optogenetic tools for intracellular signaling are not well developed. Therefore, we engineered a selective presynaptic fusion tag for photoactivated adenylyl cyclase (bPAC-Syn1a) and found its high localization at presynaptic terminals. Furthermore, an all-optical electrophysiological method revealed rapid and robust short-term potentiation by bPAC-Syn1a at brain stem-amygdala synapses in acute brain slices. Additionally, bPAC-Syn1a modulated mouse immobility behavior. These results indicate that bPAC-Syn1a can manipulate presynaptic cAMP signaling in vitro and in vivo. The all-optical manipulation technique developed in this study can help further elucidate the dynamic regulation of various cellular functions.