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Unlike replacement of solutions by microperfusion (from 1 mM to 100 mM glutamate), EPSCs recovered with much faster kinetics upon glutamate uncaging with a time constant of ~15 s, which is 10100 times faster than those estimated in isolated vesicles (Naito and Ueda, Figure 3 Despite this first success toward estimating the rate of glutamate transport in living synapses, there remain several critical concerns regarding the protocol

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And I think that theyre probably probably many of them and possibly most of them are wrong

Cell suspensions were then applied to the column, and the flow-through was collected in a new 15-ml tube