Super-Resolution Mapping of Neuronal Circuitry With an Index-Optimized Clearing Agent

摘要: Super-resolution imaging deep inside tissues has been challenging, as it is extremely sensitive to light scattering and spherical aberrations. Here, we report an optimized optical clearing agent for high-resolution fluorescence (SeeDB2). SeeDB2 matches the refractive indices of fixed that immersion oil (1.518), thus minimizing both During process, fine morphology fluorescent proteins were highly preserved. enabled super-resolution microscopy various tissue samples up a depth >100 μm, order magnitude deeper than previously possible under standard mounting conditions. Using this approach, demonstrate accumulation inhibitory synapses on spine heads in NMDA-receptor-deficient neurons. In fly medulla, found unexpected heterogeneity axon bouton orientations among Mi1 neurons, part motion detection circuitry. Thus, volumetric cleared powerful strategy connectomic studies at synaptic levels.