For many optical systems the properties and alignment of the internal apertures and pupils are not critical or controlled with high precision during optical system design, fabrication or assembly. In wide angle imaging systems, for instance, the entrance pupil position and orientation is typically unconstrained and varies over the system s field of view in order to optimize image quality. Aperture tolerances usually do not receive the same amount of scrutiny as optical surface aberrations or throughput characteristics because performance degradation is typically graceful with misalignment, generally only causing a slight reduction in system sensitivity due to vignetting. But for a large deployable space-based observatory like the James Webb Space Telescope (JWST), we have found that pupil alignment is a key parameter. For in addition to vignetting, JWST pupil errors cause uncertainty in the wavefront sensing process that is used to construct the observatory on-orbit. Furthermore they also open stray light paths that degrade the science return from some of the telescope s instrument channels. In response to these consequences, we have developed several pupil measurement techniques for the cryogenic vacuum test where JWST science instrument pupil alignment is verified. These approaches use pupil alignment references within the JWST science instruments; pupil imaging lenses in three science instrument channels; and unique pupil characterization features in the optical test equipment. This will allow us to verify and crosscheck the lateral pupil alignment of the JWST science instruments to approximately 1-2% of their pupil diameters.