Counting Synapses Using FIB/SEM Microscopy: A True Revolution for Ultrastructural Volume Reconstruction.

摘要: The advent of transmission electron microscopy (TEM) in the 1950’s represented a fundamental step study neuronal circuits. application this technique soon led to realization that number synapses changes during course normal life, as well under certain pathological or experimental circumstances. Since then, one main goals neurosciences has been define simple and accurate methods estimate magnitude these changes. Contrary analysing single sections, TEM reconstructions are extremely time-consuming difficult. Therefore, most quantitative studies use stereological three-dimensional characteristics synaptic junctions studied two dimensions. Here, count exact per unit volume we have applied new reconstruction method involves combination focused ion beam milling scanning (FIB/SEM). We show images obtained with FIB/SEM similar those TEM, but advantage permits serial large volumes tissue be generated rapidly automatically. Furthermore, compared estimates values by reconstructions. concluded not only provides actual it is also much easier faster than other currently available methods. More importantly, avoids errors introduced overcomes difficulties associated techniques.