Hi everyone,
I've been patching mice CA1 pyramidal neurons for synaptic transmission
study and have some shrinking cell problems, I cut 300um brain slices with
vibratome and store the slice in partial sucrose cutting solution (Jonas P,
osm ~320), and do recording in normal ACSF (osm 300-310) at 32 C. However,
in older mice ( > 3 weeks), after cutting the slice, the cells seem to be
shrinking (fold inward, not smooth round cells), and using the same
solutions, slices from younger mice looked very good. I don't know why this
happens, could it be pure age issue? I heard people can patch on really old
animals(2-3 months) without any problem. I think the next step i want to try
cardiac perfusion before slicing. It will be so helpful to hear from you
guys that what do you think of the potential causes for shrinking cells?
Yours sincerely,
David
