Asian carps continue to expand their range in North America, necessitating efforts to limit the spread and establishment of reproducing populations. Mississippi River Lock and Dam 19 is a high-head dam that represents a population ‘pinch-point’ as passage through the lock chamber is the only means by which fishes can complete upstream movement. As such, this location could be a pivotal control point for minimizing the spread of invasive fishes in the Upper Mississippi River and a possible candidate site for installation of deterrent measures. Our objectives were (1) to study the timing (i.e., weekly and diel) and behavior of fishes in the downstream lock approach, (2) evaluate the relation of presence in the downstream lock approach with environmental factors and lock operation, and (3) identify any upstream or downstream passage events through the lock chamber and the relation between these events and the operation of the lock. Acoustic transmitters were surgically implanted into 262 Asian carps and 216 native fishes to monitor fish activity on a telemetry receiver array deployed around and within the lock for 622 days during 2017–2018. One hundred eighty-six telemetered fish were detected in the downstream lock approach. We documented 14 upstream Asian carp passages and 10 upstream native fish passages; these passages coincided with a specific sequence of large vessel lockages. The results of this study advance our understanding of fish presence and behavior at a Mississippi River mainstem lock and dam and inform the development and testing of deterrent systems at this location or at similar pinch-point lock and dams.