LIVE WEBINAR: Tuesday, December 12, 2017 Sponsor: https://neuralynx.com/ During this webinar, Casey Stengel provides an introductory overview of Cube2: an all-in-one, wireless headstage and data acquisition system for neuronal research in freely moving, untethered subjects. He explains how Cube2 has been purpose built to enable the study of research animals in social environments by allowing multiple transmitters in the same area, pulling data to local drives. Following, Dr. John A. Wolf provides two case studies highlighting his work with Cube2 technology focused on long term chronic affects of traumatic brain injury, including the development of post traumatic epilepsy. Next, Dr Kari Hoffman presents research using the Cube wireless systems for wireless multi-site recordings in freely-behaving macaques. She discusses experimental design, common challenges and shares preliminary data demonstrating the capabilities of Cube2 and future possibilities for studies involving non-human primates. The webinar closes with a Q&A session held with the presentation group.

1. Experts discuss new configurable wireless
technologies for single unit electrophysiology
recording in small and large animals, with a focus on
applications, methodology, data outcomes, and the
impact these devices will have in research involving
untethered, naturally behaving subjects.
#LifeScienceWebinar #ISCxNeuralynx
Advancements in Wireless
Technology for Single Unit
Electrophysiology Recording

2. Advancements in Wireless
Technology for Single Unit
Electrophysiology
Recording
Casey Stengel
Founder,
President, CEO
Neuralynx
John A. Wolf, Ph.D.
Assistant Professor of
Neurosurgery
University of Pennsylvania
#LifeScienceWebinar #ISCxNeuralynx
Kari Hoffman, Ph.D.
Associate Professor
Vanderbilt University

3. InsideScientific is an online educational environment designed for life science
researchers. Our goal is to aid in the sharing and distribution of scientific
information regarding innovative technologies, protocols, research tools and
laboratory services

4. To access webinar content, Q&A reports, FAQ document, and
information on lab workshops, we invite you to join our community at
www.insidescientific.com/register

5. Neuralynx Wireless
Products and Concepts
Casey Stengel Founder, President, CEO
Neuralynx
Copyright © C.Stengel, Neuralynx & InsideScientific. All rights reserved

6. Wireless Concepts
• Why wireless?
• No expensive commutators and tethers
• Complex mazes which would tangle tethers are now possible
• Free moving large animals have new challenges:
• difficult with tethered animals
• curious NHP destroy delicate instrumentation
• large animal behavior destroys tethers
• Long term continuous recording for finding infrequent events (Epilepsy studies)
• Multiple subject, free moving, Social Interaction Studies are now possible
• Easily record in multiple environments – not just in a dedicated recording room
• Neuralynx offers Wireless products for small, medium and large animal models
• Product support is integrated into Cheetah Data Acquisition Software
• Trade offs:
– no DC or 24 bit A/D of the Digital Lynx SX and Input Boards
– fixed +/- 5mv input range
– Limited run-time based on battery mass

7. Wireless Products
for Small Subjects
• NEW Small Animal Telemetry 2 –
SAT2
• For small animals:
mice, rats, and birds
• 16 channels 20 Khz sampling rates
• Light weight:
5.2 grams with 1 hour battery
• Expansion to 32 channel in the
next version
• Continuous Wi-Fi, direct to
Cheetah for a low cost system
• Currently seeking Beta Test Sites
for Jan 2018 start
SAT2
24mm x 21mm x 4mm
(without battery)

8. Cube2 for Medium
Animals - Rodents
• Feature packed platform:
– 30 Khz 16 bit neural signal digitization
– 9 axis IMU,
– Ultrasonic Mic input,
– Digital I/O bits onboard,
– Optogenetics FLED driver available soon,
– uSD memory card recording,
– Remote power control options –
magnetic, IR, or Blue Tooth
– QuickCliptm connection system – fast and
no “bent pins”
• Folded Flex PCB – “Cube2” format for
head or backpack mounting Cube2 Base Module
(without battery or Analog Front End)

9. Cube2 for Medium
Animals - Rodents
• Multi Subject Recording
• WiFi to either Digital Lynx SX or Direct to
Cheetah
• uSD memory card recording w/o WiFi
transmission for lower power
• Battery options for 30 -> 60 min
operation – quick change connections
• 64 and 128 Channel support for rats
• Weight - 19 grams plus batteries
(5 to 15 gm)

10. Large Animal
Telemetry – Cube2
• Same Features on the Cube2 platform
• Pigs, Sheep and NHP offer larger
weight carrying possibilities for 256
channel recording and larger batteries
for extended recordings
• larger, rugged enclosures and battery
packs for safe, long term recordings
• configurable interfaces and AFEs for
specific needs of the experiment
– allows integration into the microdrive
implant
• Multi Subject Recording
• Battery options for 24 hour operation
• 64 to 256 Channels
• Options for temporary “pre-trial”
tethered operation

11. Full Accessory Line for your
specific experiments
• Cube2 Telemetry is becoming mature!
• Very Configurable
– Analog Front End (AFE) Boards – similar to
our MUX Headstages
– “Bridge Boards” to connect multiple Analog
Front End Boards to the Cube2
– Allows for configurations to meet specific
needs of the experiment & Subject
• Many Battery Options
• larger, rugged enclosures and battery packs
for safe, long term recordings
• Training Weights with accurate mass and
center of gravity
• Optogenetic FLED LED Source Driver Board
• EIBs with screw down retainers
• Remote Control Power Options
• “Shield” for heat dissipation and protection

12. "Our lab has experimented with a number
of wireless data acquisition systems for
both rodent and primate electrophysiology.
We are very pleased with the recording
quality and overall performance of
Neuralynx’s Cube2 - which we find
to be virtually indistinguishable from
traditional wired systems. We would
happily recommend this device for
any researcher seeking a reliable
and user friendly system for performing
wireless electrophysiology recording.”
Carol A. Barnes, Ph.D.
Director, ARL Division of Neural Systems,
Memory and Aging, University of Arizona
Existing
Customer
Experiences
Click To Learn More
About Cube2

13. Today, we will highlight three case studies using the
Cube2 Wireless system, which demonstrate natural
untethered behavior:
1. Rats: JA. Wolf – Freely Moving Wireless Neurophysiology in Rats
2. Pigs: JA. Wolf – Freely Moving Wireless Neurophysiology in Pigs
3. NHP: K. Hoffman – Wireless Recordings in Primates
Case Studies
View Webinar >>

14. Freely Moving Wireless
Neurophysiology in Rats
and Pigs
John A. Wolf, Ph.D.
Assistant Professor of
Neurosurgery
University of Pennsylvania
Copyright © J.A. Wolf & InsideScientific. All rights reserved

15. Wolf Lab Overview
Electrophysiological Dysfunction after TBI
– What are the mechanisms underlying cognitive dysfunction post TBI?
– Rat and Pig models of TBI
– Cognitive/spatial tasks with high density laminar hippocampal/cortical
recordings
– Does TBI disrupt organization of Neuronal Ensembles?
How Does TBI lead to Epileptogenesis and PTE?
– Chronic video/EEG as well as depth probes in the HC of pigs
– Comparison of different TBI modes

16. Laminar Structure w/Si Probes (Cambridge)
Increase in Firing Rate
with 32 edge channelssecured to asmall high-resolution microdrive (Cambridge NeuroT
30 kHzwirelessly.
Chronica
with wi
recording
High-reso
with mo
stereotax
(Cambrid

17. Laminar Structure w/Si Probes (Cambridge)
Increase in Firing Rate
with 32 edge channelssecured to asmall high-resolution microdrive (Cambridge NeuroT
30 kHzwirelessly.
Chronica
with wi
recording
High-reso
with mo
stereotax
(Cambrid
1. Laminar structure important feature of
HC input/ organizing oscillations
2. Laminar probes (64 channels, 2 shanks)

18. Laminar Structure

19. Rat Wireless – HC Units
• CUBE64
• 32 channel Buzsaki style probe
(less laminar info)
• Rat Hippocampus
Hippocampal Units
Pyramidal Cell
y dysfunction
ared to sham
ggest that FPI

20. Example Amygdalar/PFC/vHC Recordings
• 32-channel silicon probe, 2 shanks in BLA • Bipolar with units in HC/PFC • Low level of noise

21. Noise Comparison
(representative rats)
[TOP]
Analog Tether
[BOTTOM]
Wireless

22. • References
– Don’t tie Ref and Ground together (increases noise 2X)
– Non- selectable, so choose carefully
– Ventricular ref. best in our hands
• Compression algorithm on SD card storage helps w/space
– Managing it afterward for long recordings takes some planning
• Custom AFEs can help offload weight
• Acclimate rats for a few days to CUBE weight if not backpacked
– 60 mins for reasonable weight for rat head mounted
– Single cell battery for 25 mins fine for many behavioral experiments
General Technique Notes

23. Post Traumatic Epilepsy
1. Injury 2. Implantation
3. Recordings / Monitoring4. Neuropathology
ECoG
HC
Layers

24. Sagittal Section of
7mm off Midline
• LFB/CV

25. Sagittal Section of Pig HC
Sagittal Section
of Pig HC

26. Vector EDGE-32 Silicon Probe

27. Large Animal Custom
Enclosure System (LACES)
• CUBE2
• 64-128 channel Wireless
• Analog Front End for A/D
• INTAN Chips (30kHz
Sampling)
• SD Card Writing (256Gb)
• 16-24 hr battery life
• Swap cards/batts every
other day
BLACK DELRIN ENCLOSURE COVER WITH TRANSLUCENT WINDOW
24 HOUR BATTERY PACK
ALUMINUM ENCLOSURE BODY BRIDGE CABLE
BRIDGE CABLE PROTECTION
ALUMINUM ENCLOSURE ADAPTER
CABLE ADAPTER BOARD
AFE
ADAPTER PCB
HEADCAP CHAMBER
ROCKER LATCH
BODY CABLE PASS THROUGH
D
C
BB
C
D
8 7 6 5 4 3 2 1
E
F
E
F

28. Pig Chronic Wireless Recordings (Homecage)

29. WAP WAP
Cage 1 Cage 2 Cage 3 Cage 4
Pig Wireless Epilepsy Monitoring Unit

30. WAP WAP
Cage 1 Cage 2 Cage 3 Cage 4
Pig Wireless Epilepsy Monitoring Unit

31. Wireless Behavior
CA1 Pig Place Cell

32. Click To Access The
Webinar Recording

33. Example LFP and Units – Pig HCExample LFP and Units – HC
• Wideband LFP • Behavioral Space

34. Co-Is
• Victoria Johnson, M.D., Ph.D
• Isaac Chen, M.D.
• D. Kacy Cullen, Ph.D.
• Brian Litt, M.D.
Acknowledgements
Funding
• DoD CDMRP
• VA RR&D
• NINDS
• CURE
Wolf Lab
• Alexandra Ulyanova, Ph.D.
• Chris Adam
• Carlo Cottone, Ph.D.
• Robin Russo
• Kim Gagnon

35. Wireless Recordings in
Primates
Copyright © K. Hoffman & InsideScientific. All rights reserved
Kari Hoffman, Ph.D.
Associate Professor of
Psychology
Vanderbilt University

36. Flex Recording Collaborators
• Chris Lewis & Pascal Fries, ESI
Frankfurt
• Thilo Womelsdorf, Vanderbilt
University
Acknowledgements
Funding
• Brain Canada
• Krembil Foundation
• Alfred P. Sloan Fellowship
• NSERC DG and CREATE
VSA
• Canada Foundation for
Innovation
Wireless Recordings
• Patricia Sayegh
• Omid Talakoub
• Special thanks to Andrea
Gomex Palacio Schjetnan and
Hjalmar Turesson

37. Why were we interested in wireless recordings?
• Interest in the neural basis of learning and memory
• Traditionally done with head-restrained macaques viewing
visual stimuli on a monitor

38. Preparation:
Chronic implantation
• Trec Tetrodes in a microdrive
*retrofit
• 16-ch ‘Flex probes’ (collab. P. Fries,
C. Lewis, now Blackrock)
• Neuralynx custom adapter boards
(Electrode interface boards)

39. Preparation:
Chronic implantation
• Trec Tetrodes in a microdrive
*retrofit
• 16-ch ‘Flex probes’ (collab. P. Fries,
C. Lewis, now Blackrock)
• Neuralynx custom adapter boards
(Electrode interface boards)
• Early thermoplastic, 3-D printed
holders
• Later 3-D printed holders on
custom implant base (S. Frey, B.
Hynes)
Omid Talakoub
Kari Hoffman
Thilo Womelsdorf

40. Omid Talakoub
Kari Hoffman
Thilo Womelsdorf

41. Omid Talakoub
Kari Hoffman
Thilo Womelsdorf

42. Omid Talakoub
Kari Hoffman
Thilo Womelsdorf

43. Cube/Cube2 in Action

44. Cube/Cube2 in Action

45. Cube/Cube2 in Action

46. Early Implications
• Immediate feedback on free-behaving neural correlates
• With proper connections, referencing we saw healthy LFP
• With write-to-card, hybrid approaches, can do overnight, ~6 hours
• New domain of experiments…for primate electrophysiologists

47. Casey Stengel
Founder,
President, CEO
Neuralynx
John A. Wolf, Ph.D.
Assistant Professor of
Neurosurgery
University of Pennsylvania
Kari Hoffman, Ph.D.
Associate Professor
Vanderbilt University
For additional information on the products
and applications presented during this
webinar please visit www.neuralynx.com
Thank You
#LifeScienceWebinar #ISCxNeuralynx