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-- 1 Voltage-Sensitive Dye Optical Imaging Vassiliy Tsytsarev Department of Anatomy and Neurobiology.

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Presentation on theme: "-- 1 Voltage-Sensitive Dye Optical Imaging Vassiliy Tsytsarev Department of Anatomy and Neurobiology."— Presentation transcript:

1 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 1 Voltage-Sensitive Dye Optical Imaging Vassiliy Tsytsarev Department of Anatomy and Neurobiology University of Maryland School of Medicine E-mail: tsytsarev@umaryland.edutsytsarev@umaryland.edu

2 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 2 Principles of Voltage-Sensitive Dye Optical Imaging RH-795 chemical structure Dye interactions with the lipid bi- layer Unbound to the lipid bilayer dye molecules are not fluorescent Bound dye molecules are fluorescent (after: Grinvald et al. 2001. In-vivo Optical imaging of cortical architecture and dynamics)

3 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 3 Main Parts of the VSD Imaging Setup Figure: Schematic of measurement CCD camera Excitation Filter (620+ 20 nm) Emission Filter (695nmLP) Dichroic mirror LFP E-stim 4AP Figure 2 Schematics of experimental setup Emission Filter (695 nm LP) CCD camera Dichroic mirror Electrical stimulation LFP recording Intracortical Injection Excitation Filter (620± 20 nm Light Shutter

4 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 4 How VSD works? (After: Tsytsarev et al, 2008 Imaging cortical electrical stimulation in vivo: fast intrinsic optical signal versus voltage-sensitive dyes) Electrode 1 mm Frame = 10 ms VSD vs fast IOS

5 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 5 VSD Imaging of Tonotopicity 1 kHz 5 kHz 7 kHz Color-coded tonotopic map of sound frequencies showing the organization of subfields of the auditory cortex (After: Tsytsarev et al, 2009. Optical imaging of interaural time difference representation in rat auditory cortex) 1 kHz 5 kHz 7 kHz

6 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 6 Voltage-Sensitive Dye Optical Imaging of the Auditory Cortex Where is in the brain? “Where”

7 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 7 Interaural Time Difference (ITD) Binaural click produces the sensation of a sound source to the left, in front of, or to the right of the animal. The angle is determined by the Interaural Time Difference (ITD) (After: Tsytsarev et al, 2009. Optical imaging of interaural time difference representation in rat auditory cortex)

8 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 8 Imaging of the ITD Representation sound stimulus onset Click leading: Red: contralateral Green: ipsilateral Blue: (ITD = 0) (After: Tsytsarev et al, 2009. Optical imaging of interaural time difference representation in rat auditory cortex)

9 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 9 Conclusion l The virtual map of auditory space in rat auditory cortex does not exist but l Patterns of neural activity recorded in response to binaural click presentations can encode ITD

10 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 10 Vibrissae System: Optical Imaging Research The main goal: to create a functional map of the directional sensitivity of the barrel field using Voltage-Sensitive Dye Optical Imaging

11 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 11 Vibrissae System: Angular Selectivity in the Barrel Field? 1 mm Stimuli Arrangement (color coded) Whisker’s Stimulator VSD optical patterns evoked by different stimuli

12 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 12 Studying of the Cortical Representation of Whisker Directional Deflection Using Voltage-Sensitive Dye Optical Imaging Barrel Field VSD Pseudocolor Pattern in the Region of Interest Direction of the Deflection Magnetized Whisker inside the Stimulator

13 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 13 Barrel Field Voltage-Sensitive Dye Optical Imaging Histology (Cytochrome-c oxidase) Extracted brain Barrel field Brain opening and area of recording 0.15 0 -0.15 Δ F/F(%) 0 20 time, ms Integrated signal in response to different stimuli

14 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 14 Activity Centers in the Single Coordinates System Borders of the Activity Patterns after thresholding 0.5 mm

15 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 15 Disposition of Centers of Mass of Activity Patterns Stimulus 1 Stimulus 2 Stimulus 3 Stimulus 4

16 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 16 Acknowledgments  Drs. Shigeru Tanaka, Hidenao Fukuyama, Kazuyuki Imamura, Ayako Ajima, Hisayuki Ojima, Minoru Kimura and Jerom Ribot – Brain Science Institute of RIKEN and Kyoto University, Japan  Dr. Sonya Bahar, Director, Center for Neurodynamics, University of Missouri at St. Louis  Daisuke Takeshita Douglas Joseph Brumm and Dr. Micheal Hoffman – Center for Neurodynamics, University of Missouri at St. Louis  Song Hu, Junjie Yao, Li Li – Ph.D. student of the Washington University in St. Louis  Drs Konstantin Maslov and Lihong Wang – Department of Biomedical Engineering, Washington University in St. Louis

17 http://oilab.seas.wustl.eduhttp://oilab.seas.wustl.edu -- 17 Thank you very much for your attention

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