Abstract
Electroretinography (ERG) is a foundational method for assessing visual system physiology, but accurate ERG can be time- and labor-intensive, often involving manual adjustment of the wavelength and intensity of light stimuli and real-time comparison of physiological responses to inform those adjustments. Furthermore, current approaches to ERG often require expertise beyond that necessary for the electrophysiological preparation itself. To improve both the efficiency and accessibility of ERG, we designed an automated system for stimulus presentation and data acquisition. Here, we test this novel system’s ability to accurately assess spectral sensitivity in the well-characterized visual system of the crayfish Procambarus clarkii using three approaches: the first, based on response magnitude, maximizes efficiency; the second is a well-established method we use to further validate our efficient approach’s accuracy. Third, we explore the potential benefits of extensible automation using a method assessing the interplay between temporal acuity and spectral sensitivity. Using our system, we are able to acquire accurate results in ERG experiments quickly (testing the entire visible spectrum in 8 min, 30 s using our response magnitude approach). Moreover, data collected via all three methods yielded results consistent with each other and previous work on P. clarkii.
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Availability of data and materials
The data that support the findings of this study are available from the corresponding author, LTH, upon reasonable request.
Code availability
All code involved in the automated ERG system is open-source and can be found at lukehavens.com/projects/electroretinography/. Code performing common functions such as visual pigment curve-fitting and statistical analyses are available from the corresponding author, LTH, upon reasonable request.
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Acknowledgements
For their advice on this project, the authors thank Christine Bedore and Dan Chappell. LTH also thanks Dick Vogt for initial training in the field of electrophysiology.
Funding
Support for the authors was provided, in part, by IOS Award no. 1457148 from the National Science Foundation (to DIS), as well as a Magellan Scholar Award from the University of South Carolina and a Science Undergraduate Research Fellowship from the South Carolina Honors College (both to LTH).
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LTH designed and built the electroretinography apparatus, conceived of and programmed the automated electroretinography protocols, and collected spectrally influenced temporal acuity and sensitivity-converted response data. ACNK collected response magnitude data. DIS oversaw the project. All authors contributed to analyzing data and drafting the manuscript.
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We followed all applicable rules and regulations for the care of crustaceans in the United States.
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Havens, L.T., Kingston, A.C.N. & Speiser, D.I. Automated methods for efficient and accurate electroretinography. J Comp Physiol A 207, 381–391 (2021). https://doi.org/10.1007/s00359-021-01476-4
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DOI: https://doi.org/10.1007/s00359-021-01476-4