Title: Visual masking with optical imaging of intrinsic signal, twophoton calcium imaging, and single unit
1Visual masking with optical imaging of intrinsic
signal, two-photon calcium imaging, and single
unit recording.
MNTP2009 Optical Imaging Modality
- Anastasia Gorbunova, MA (University of Arizona)
- Modality instructor Dr. Justin Crowley
- Training Assistant Corey Flynn
2Visual 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
doctor
DOCTOR
doctor
R. Fox. (1998). In Handbook of Sensory
Physiology. VIII. Perception.
3Optical 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
4Stimuli
1.
2.
Time
4.
3.
Time
5Stimuli
5.
6.
7.
8.
6Intrinsic 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
-
Courtesy of David E. Whitney
7Experiment 2 F09-051
1 Block
-
8Experiment 2 F09-051
8 s
8 s
8 s
100 ms
100 ms
100 ms
9Experiment 2 F09-051
1 frame 500 ms
10Extracellular Single Unit Recordings
- Exposed visual cortex of a ferret (P80-130)
- Carbon fiber electrode
- 20kHz signal digitization
-
11Extracellular Single Unit Recordings
B
H
H
V
V
B
SQP
SNP
SNP
SQP
H-B
H-B
V-B
H-V
V-H
V-H
H-V
V-B
H, horizontal grating B, blank V, vertical
grating SQP, square-wave plaid SNP, sine-wave
plaid
122-photon calcium imaging
- Oregon green BAPTA-1AM (labels neurons and
astrocytes) - Sulphorhodamine-101 (labels astrocytes)
- Microscope Zeiss LSM 510 Meta NLO with MIRA900
Ultrafast TiSapphire laser - 20x NA1 objective
13Calcium imaging
142-photon calcium imaging
3 cells ?F/F for 700 frames normalized over 10
blocks
15Conclusions
- 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
16Acknowledgements
Dr. Justin Crowley Corey Flynn David
Whitney Sruthi Chintakunta Alberto Vazquez
Dr. Seong-Gi Kim Dr. Bill Eddy Tomika
Cohen Rebecca Clark
Thank you!