NINDS 24-000023 NOTICE OF INTENT

The National Institute of Neurological Disorders and Stroke (NINDS), Synaptic Transmission Section (STS) performs single-molecule localization imaging of fluorescently labeled proteins for experiments. This multi-color imaging technique allows 1) imaging of the high-resolution structure of the cell membrane, 2) localizing individual molecules within cells to understand their nanoscale distribution at a localization precision of single nanometers, and 3) tracking the dynamics of a variety of proteins and other microscopic during vesicle exocytosis and endocytosis in the cells. These precise, quantitative, yet lengthy experiments require an automated perfusion system that allows seamless buffer exchange at a steady rate over a long duration allowing multi-color imaging.

The synaptic Transmission Section combines physiological and cell-biological approaches to understand the dynamic modulations of the cell membrane. It investigates the role of various proteins during vesicle exocytosis and endocytosis in living cells. Some of the driving questions include how protein localization and clustering at the nanometer scale can lead to emergent alterations at a larger scale, such as the changes in the plasma membrane during vesicle docking, curvature generation, endocytosis, and so on. Our mission is to characterize these processes with a super-resolution imaging technique, where we can simultaneously visualize multiple proteins through single-molecule fluorescence imaging and then determine the underlying mechanisms. This approach requires hours, if not days, of continuous imaging of numerous proteins via MINFLUX nanoscopy, where each step requires seamless buffer exchange. Therefore, it is critical to automatize the entire process to reduce the experimental dead time and human errors, such as introducing localization noise while handling the imaging setup during an ongoing experiment. Further, in such experiments, active flow control is essential as the perfusion systems based on gravity can lead to inconsistent flow rates comparable to the experimental scale. 

The contractor shall deliver within 10 weeks after receipt of the order. The contractor shall provide inside delivery and installation of the equipment within 70 days after receipt of the order. The site location address is Building 35, Room 2BC-909, 35 Convent Drive, Bethesda, MD 20892-3706.

All services performed on this equipment shall be provided by certified and factory-trained technicians. Technicians shall provide training on handling the instrument for regular usage. They will diagnose the systems on-site via remote access if needed. The contractor shall provide any required replacement components due to on-site malfunctions within the one-year warranty period.