Available Technology

Deep Tissue Probe Imaging with Quantum Dot Tips

NRL and its collaborators have developed a more robust fluorescent labeling of standard borosilicate glass pipettes allowing their 2P visualization far deeper within complex tissues. Recording pipettes are coated with luminescent semiconductor quantum dots (QDs) and used to visualize the probe under 2P microscopic illumination, enabling simultaneous electrophysical recordings and cellular binding measurements.
Patent Abstract: 
Electrophysiological recordings at the single cell level within complex matrices such as tissues, organs or even in vivo is a fundamental technique for neuroscience, cardiology and electrophysiology. One significant hurdle to accomplishing this is visualizing the patch clamp or pipette tip deep within these dense tissues. Currently, these studies are typically accomplished by patch clamp visualization using two-photon (2P) imaging of organic dyes such as fluorescein that are continuously expelled from the pipette tip. Limitations of this technique include their relatively low 2P action cross-section, their susceptibility to photobleaching, and dye accumulation causing increased background, especially after multiple descents. To address this, NRL and its collaborators have developed a more robust fluorescent labeling of standard borosilicate glass pipettes allowing their 2P visualization far deeper within complex tissues. Recording pipettes are coated with luminescent semiconductor quantum dots (QDs) and used to visualize the probe under 2P microscopic illumination, enabling simultaneous electrophysical recordings and cellular binding measurements. This approach has been successfully demonstrated for targeting cells deep within rat and mouse brain in vivo. Using a series of green, yellow and red emitting QDs, experiments were conducted on hippocampal DS-red-labeled cholecystokinin positive interneurons and GFP-expressing parvalbumin-positive interneurons. GFP expressing neurons were even successfully electroporated in vivo at > 750 ┬Ám using 625 QD coated peptides.
Benefits: 
applicable for cells deep within a tissue, - continuous visualization and electromonitoring over extended periods of time, - applicable in both in vivo and ex vivo experiments, - simultaneous electrophysical recordings and cellular binding measurements - facilitates improved pipette imaging and electrophysiological recording in many different types of tissue samples.
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