Dynamic neuroimaging of retinal light responses using fast intrinsic optical signals.
ABSTRACT Transient intrinsic optical responses associated with neural activation offer an attractive strategy for dynamic imaging of neural activity, and may provide a noninvasive methodology for imaging of retinal function. Here we demonstrate the feasibility of near infrared imaging of fast intrinsic optical changes in isolated frog retina activated by visible light. Using a photodiode detector in a transmitted light geometry, we routinely measured dynamic transmitted optical responses in single passes, at the level of one part in 10(4) of background light. Rapid CCD image sequences acquired with transmitted light (bright field) illumination disclosed larger fractional responses and showed evidence of multiple response components with both negative- and positive-going signals with different timecourses. Dark field imaging further enhanced the contrast and sensitivity of optical measures of neural activation. High-resolution imaging disclosed optical responses in single pixels often exceeding 5%, of background light, allowing dynamic imaging at the resolution of single cells, in single passes. Fast optical signals are closely related to identified response components of the electroretinogram. Optical responses showed complex but consistent spatial organization from frame to frame. Our experimental results and theoretical analysis suggest that the optical responses may result from dynamic volume changes corresponding to ion and water flow across the cell membrane, directly associated with the electrophysiological response.
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ABSTRACT: Intrinsic optical signal imaging (IOS) promises a noninvasive method for high resolution examination of retinal function. Using freshly isolated animal retinas, we have conducted a series of experiments to test fast IOSs which have time courses comparable to electrophysiological kinetics. In this article, we demonstrate the feasibility of in vivo imaging of fast IOSs correlated with retinal activation in anesthetized frog (Rana Pipiens). A rapid (68,000 lines/s) line-scan confocal ophthalmoscope was constructed to achieve high-speed (200 frames/s) near infared (NIR) recording of fast IOSs. By rejecting out-of-focus background light, the line-scan confocal imager provided enough resolution to differentiate individual photoreceptors in vivo. With visible light stimulation, NIR confocal images disclosed transient IOSs with time courses comparable to retinal ERG kinetics. High-resolution IOS images revealed both positive (increasing) and negative (decreasing) light responses, with sub-cellular complexity, in the activated retina.SPIE Optical Engineering + Applications; 09/2011
Edited by Ruikang K. Wang, Valery V Tuchin, 01/2013: pages 559-580; Taylor & Francis.
In Vivo Optical Imaging of Brain Function., 2 edited by Frostig RD, 01/2009: chapter 5; CRC Press., ISBN: 978-1-4200-7684-4