Current preclinical eye and eye-brain research often includes the collection of geographical data, such as fluorescence imaging, and structural data obtained using techniques such as optical coherence tomography. However, seldom are functional measurements of the eye measured using electroretinography (ERG) as it has traditionally been viewed as a difficult technology to implement. Fortunately, innovation in the tools and techniques to measure ERG in rodent subjects now offer a dramatic decrease in complexity for the scientist and thus pave a new way forward for comprehensive study of eye function and associated disease. In this webinar, sponsored by Phoenix Research Labs, scientists discuss the essential elements of measuring retina function in rodents, highlighting key technology, methodology and best practices that will lead to accurate and repeatable results in the lab. Dr. Peter Lundh von Leithner, PhD from University College London shares the importance of electrophysiology to his research and how he has optimized equipment for his particular studies. Dr. Christine van Hover, Ph.D. also reviews the four step process uniquely available with the Phoenix Research Labs technology. Key Topics: * the importance of electroretinography in eye research * key differences when testing the rodent versus human eye -- optimizing system settings for the rodent eye including electrodes, dark adaptation and the two classes of cones * obtaining quality data without a Faraday cage * fundamental elements of data export, display and analysis * the power of “total information” -- how adding functional measurements to each retinal evaluation can reveal critical information

1. Measuring Retinal
Function with ERG in Rodents
Sponsored by:
Christine
van Hover, PhD.
Field Application Specialist,
Phoenix Research Labs
Peter Lundh von
Leithner, PhD.
Research Scientist,
University College London

2. 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.

3. Thank you to our event sponsor

4. Copyright 2016 Phoenix Research Labs and InsideScientific. All Rights Reserved.
Christine van Hover, PhD.
Field Application Specialist,
Phoenix Research Labs
Sponsored by:
Measuring Retinal
Function with ERG in Rodents

5. Phoenix Research Labs Electroretinography

6. What is ERG?
Measuring the
function of retinal
cells by detecting their
electrical activity in
response to flashes of
light

7. Phoenix has developed technology to enable a four step
process
1. Prepare animal
2. Establish corneal contact
3. Acquire data
4. Analyze and present data
Ganzfeld ERG Focal ERG

8. • Dark lab technology
enables easy set up and
dark adaptation
• Needle electrodes are
easy to place and
secure
• ERG systems
specifically designed
for rodent eyes
1. Prepare Animal

9. • Near infrared light
allows image-guided
alignment with solid
contact electrode
• Unique Maxwellian view
allows uniform
illumination of the eye
• Live chart mode shows
real time coupling with
the cornea
2. Establish corneal
contact

10. 2. Establish corneal
contact
• Near infrared light
allows image-guided
alignment with solid
contact electrode
• Unique Maxwellian view
allows uniform
illumination of the eye
• Live chart mode shows
real time coupling with
the cornea

11. 3. Acquire Data
Before
After
• Simple interface to
select protocol and
collect data
• Clean algorithm
removes 50 Hz or 60
Hz noise
• Allows detection of
small differences
• Fewer light flashes
• No Faraday cage
needed

12. • LabScribe calculates A
and B waves
• Export graph and table
of results
• Oscillatory potentials
can be examined
separately
• Automatic waterfall
provides effective
review
4. Analyze and
present data

13. 4. Analyze and
present data
• LabScribe calculates A
and B waves
• Export graph and table
of results
• Oscillatory potentials
can be examined
separately
• Automatic waterfall
provides effective
review

14. Copyright 2016 P. Lundh von Leithner, Phoenix Research Labs and InsideScientific. All Rights Reserved.
Peter Lundh von Leithner, PhD
Research Scientist,
University College London
Measuring Retinal
Function with ERG in Rodents

15. Measuring Retinal Function using
focal ERG in Rodents
1. Who we are and what we do
2. How we generate Electroretinography (ERG) data
3. How we evaluate ERG data sets in the context of
pharmacological assays
4. Study examples

16. Institute of Ophthalmology UCL
goo.gl/q5zEbw
Moorfields Eye Hospital
tinyurl.com/hmz7um9
University College London
goo.gl/gy5f

17. Translational Vision Research (TVR) Lab
• Professor Dave Shima
• Identify novel functions of vascular
growth factors (VGF and their
potential efficacy as therapeutic
targets
• VEGF role in neuron migration in
development and as a
neuroprotectant in the adult retina
• Present efforts are focused on
determine the neurorotectiove roles of
VGF in models of DR and Glaucoma

18. TVR In Vivo
• Fluorophotometry
• Perfusion assays
• Electroretinography
• Fluorescence funduscopy
• Scanning Laser Ophthalmoscopy
• Optical Coherence Tomography

19. Electroretinography (ERG) is a physiological assay
used to monitor retinal function of the retina. Specifically,
the function of the light-sensitive cells of the eye, the rods
and cones, and their connecting ganglion cells in the
retina are examined.

20. Preclinical electroretinography
• Developing physiological assays to assess the degree of retinal
function in models of vision
• Phenotyping of rodent models with including visual signal processing
• Monitoring sensory performance during and after therapeutic
interventions.

22. Amplification of signal

23. Animal Handling
Ketamine+Xylazine
Hypnovel+Domitor+Sublimaze
Isoflurane
Tropicamide 1%
Phenelypherine 1%

24. Photopic

25. Photopic
25

26. Stimulator settings

27. Scotopic
27

28. Scotopic
Micron Discover settings
28

31. 31

33. 33

37. fERGFFA

38. References
• Doyle, S. L., López, F. J., Celkova, L., Brennan, K., Mulfaul, K., Ozaki, E., et al. (2015).
IL-18 Immunotherapy for Neovascular AMD: Tolerability and Efficacy in Nonhuman Primates.
Investigative Ophthalmology & Visual Science, 56(9), 5424–7.
• Nagai, N., Lundh von Leithner, P., Izumi-Nagai, K., Hosking, B., Chang, B., Hurd, R., et al. (2014).
Spontaneous CNV in a novel mutant mouse is associated with early VEGF-A-driven angiogenesis
and late-stage focal edema, neural cell loss, and dysfunction. Investigative Ophthalmology & Visual
Science, 55(6), 3709–3719.
• Tanimoto, N., Sothilingam, V., & Seeliger, M. W. (2012). Functional Phenotyping of Mouse Models
with ERG. (Vol. 935, pp. 69–78). Totowa, NJ: Humana Press.
• Nusinowitz, Steven, et al. "Electrophysiological testing of the mouse visual system." Systematic
Evaluation of the Mouse Eye: Anatomy, Pathology, and Biomethods (2002): 320-344.

39. Thank you!
• Prof. Dave Shima
• Peter Lundh von Leithner
• Dr Vivian Lee
• Dr Brett Hoskins
• Dr Ewa Kubala
• Joanna Holeniewska
• Shannon Conder

40. Peter Lundh von Leithner, PhD
Peter.lundh@ucl.ac.uk
Christine van Hover, PhD
cvanhover@phoenixreslabs.com
Thank You!
If you have questions for the presenters
please contact them by email.
For additional information on the solutions
presented in this webinar please visit:
http://phoenixreslabs.com