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In brief, brains were postfixed in 4% paraformaldehyde (Fisher #AC169650010) in 0.1 m phosphate buffer at +4C for 24 h then equilibrated in 30% sucrose (Fisher S5-3) in 0.1 m PBS (Fisher #BP399-20) at +4C before slicing on a freezing microtome (Leica) in a 1:12 series of 40-m-thick coronal sections

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So we usually do not do a single intradiscal injection into one disc