I work with brain slices using the multiclamp 700B in whole cell mode. The cells start looking unhealthy within 1 hour of the surgery, with nuclei becoming distinctly visible under the DIC microscope. I am still able to get Gigaohm seals with these cells but the break-ins are mostly spontaneous or unsuccessful and the cells do not give any action potentials. Can someone please help with this? (I use 1.5/0.86 Sutter pipettes with resistance about 5-6 Mohms).
improving health of brain slices & breaking in