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MNTP2009: Optical Imaging Modality. Anastasia Gorbunova, MA (University of Arizona) Modality instructor: Dr. Justin Crowley Training Assistant: Corey Flynn. Visual masking with optical imaging of intrinsic signal, two-photon calcium imaging, and single unit recording. Visual Masking.

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Presentation Transcript
slide1

MNTP2009: Optical Imaging Modality

Anastasia Gorbunova, MA (University of Arizona)

Modality instructor: Dr. Justin Crowley

Training Assistant: Corey Flynn

Visual masking withoptical imaging of intrinsic signal, two-photon calcium imaging, and single unit recording.

visual masking
Visual Masking
  • “Destructive interaction or interference among transient stimuli that are closely coupled in space or time”

Forster & Davis (1984)

J Exper Psych: Learning, Memory & Language

Time

DOCTOR

500 ms

#######

doctor

DOCTOR

doctor

50 ms

######

doctor

500 ms

R. Fox. (1998). In Handbook of Sensory Physiology. VIII. Perception.

optical imaging modality
Optical Imaging Modality
  • Optical imaging of intrinsic signal

- wide spatial field

- temporal resolution

slow hemodynamic dependent

faster light scattering properties of tissue

  • Single unit recordings

- best temporal resolution

  • Two-photon Calcium imaging

- distinct neurons, fine time course

stimuli
Stimuli

1.

2.

Time

Time

4.

3.

Time

Time

stimuli1
Stimuli

5.

6.

8.

7.

Time

Time

intrinsic signal oi
Intrinsic Signal OI
  • Exposed visual cortex of a ferret (P80-130)
  • Illumination by ~700 nm light
  • Visual stimulation (4 degree bars)
  • CCD camera (100 ms data frames)
  • Computer digitization

8000 ms

Courtesy of David E. Whitney

experiment 2 f09 051
Experiment 2: F09-051

1 Block

ISB 3 s

Time

Blank stimulus 8.1 s

ISB 3 s

8 s

Stimulus 2

0.1 s

Inter-Stimulus Blank 3 s

8 s

Stimulus 1

0.1 s

-

=

experiment 2 f09 0511
Experiment 2: F09-051

8 s

8 s

8 s

100 ms

100 ms

100 ms

experiment 2 f09 0512
Experiment 2: F09-051

8 s

8 s

8 s

1 frame: 500 ms

100 ms

100 ms

100 ms

extracellular single unit recordings
Extracellular Single Unit Recordings
  • Exposed visual cortex of a ferret (P80-130)
  • Carbon fiber electrode
  • 20kHz signal digitization
slide11

Extracellular Single Unit Recordings

V

B

V

B

H

H

SQP

SNP

SNP

SQP

H-V

H-B

H-V

V-H

H-B

V-H

V-B

V-B

H, horizontal grating; B, blank; V, vertical grating; SQP, square-wave plaid; SNP, sine-wave plaid

2 photon calcium imaging
2-photon calcium imaging
  • Oregon green BAPTA-1AM (labels neurons and astrocytes)
  • Sulphorhodamine-101 (labels astrocytes)
  • Microscope: Zeiss LSM 510 Meta NLO with MIRA900 Ultrafast Ti:Sapphire laser
  • 20x NA1 objective
2 photon calcium imaging1
2-photon calcium imaging

3 cells: ΔF/F for 700 frames normalized over 10 blocks

conclusions
Conclusions
  • It is possible to study masking in animal models using simple visual stimuli
  • Very brief presentation of static gratings produces sufficient activation detectable by optical imaging of intrinsic signal
  • Orthogonal mask disrupts that activation
acknowledgements
Acknowledgements

Dr. Justin Crowley

Corey Flynn

David Whitney

Sruthi Chintakunta

Alberto Vazquez

Dr. Seong-Gi Kim

Dr. Bill Eddy

Tomika Cohen

Rebecca Clark

Thank you!