Project Summary This is a supplement request for purchasing two instruments. One is a high-pressure freezing apparatus which is needed to preserve the cell or tissue in vitrified state for cryoFIB and cryoET experiments. This approach is well documented in the literature to be effective. The proposed Leica ICE+ Pearl have an added advantage that we can manipulate the sample with external electrical or optical stimuli immediately before freezing. This will allow us to subject the cell sample in different physical conditions mimicking physiological conditions after which the sample will be frozen to capture its structure for cryoET observation. The second instrument is a micropatterning device to write on the EM grid to make the cells spreading more readily across the grid square rather than the grid bars. Our preliminary experiments demonstrate the effectiveness of this approach.