Immunofluorescent staining was performed to assess the impact of dexmedetomidine on neuronal cytoskeletal growth in neurons grown for 7 days post-culture. Fixation and staining were done as previously described for neurofilament antibody34 (link). Cells were imaged on a Zeiss Axio Observer Z1 microscope (Zeiss Corp.) using a 20×/0.8 DICII objective. Fifteen to twenty neurons per biological replicate (n = 12–13) were quantified.
A procedure similar to the one previously outlined was followed to evaluate the impact of DEX on synaptic network assembly in neurons grown for 7 days post-culture using synaptophysin and PSD-95 antibodies as previously described34 (link). Cells were imaged on a Zeiss Axio Observer Z1 microscope (Zeiss Corp.) using a 63×/1.4 oil DICII objective 15–20 neurons per biological replicate (n = 8–10) were quantified.
A procedure similar to the one previously outlined was followed to evaluate the impact of DEX on synaptic network assembly in neurons grown for 7 days post-culture using synaptophysin and PSD-95 antibodies as previously described34 (link). Cells were imaged on a Zeiss Axio Observer Z1 microscope (Zeiss Corp.) using a 63×/1.4 oil DICII objective 15–20 neurons per biological replicate (n = 8–10) were quantified.
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