Pyramidal neurons of CA1 neuron in organotypic slice

This is a maximal intensity projection of CA1 pyramidal cell transfected with plasmid with the reporter GFP using single cell electroporation technique. In this particular case the organotypic slices were prepared from p5-7 pups in a tissue chopper (McIlwain). And maintained in MEM bases media with added glutamax with a change in 2 alternative dyas at 37°C and 5% CO₂ for 4 days in-vitro (DIV) before electroporating with a glass pipette of 7-10mΩ resistance by applying 4 square pulses of -ve voltage of -2.5 V, for 25 ms duration at 1 Hz frequency. Then maintained in the same medium for one four more days before fixing them in PFA(4‰ v/v) and amplified with anti-GFP antibody (ab290, 1:1000 dil) and imaged in SP2 confocal microscope (Leica) for z-stack.