In this section, we will develop novel technologies and approaches to increase the success rate of integral membrane protein structure determination by x-ray crystallography and cryo-electron microscopy (cryo-EM). The focus will be on sample preparation. Three of the four methods described depend on sequence optimization of the target membrane protein, either by (1) identifying and expanding upon regions of genomic sequence space most likely to harbor proteins amenable to in-depth structural analysis; (2) by producing libraries of random mutants of the target protein, and selecting those with increased probability of success in structure determination; (3) by genetically engineering chimeras between integral membrane proteins and soluble chaperones to promote either crystallization in lipidic cubic phase, or large molecular-weight assemblies for cryo-EM structure determination. In (4), we describe high-throughput screening methods to identify appropriate samples for high-resolution cryo-EM structure determination.