Course "Electrophysiological Recordings and Optical Imaging in Neuroscience"

Abstract

Neurons are excitable cells that express themselves electrically. These signals - changes in membrane potential caused by ions moving through channels - take place on a millisecond timescale and over a range of millivolts, involving tiny currents of pico- and nano ampères. The nature of these signals and their time scale impose severe restrictions on the experimental approaches for studying the physiology of neurons. The two most suitable methods are electrophysiology and imaging. However, the technical nature of these disciplines makes them inaccessible to many neurobiologists. This invisible barrier is an obstacle to the development of neuroscience. The goal of the course is to present in a simple way the principles, capabilities and limitations of electrophysiological and imaging techniques. It will provide a theoretical grounding of each technique, and illustrate each using real examples, in many cases produced in the laboratory of the instructor. The course will consist of two modules: the first devoted to the study of the bases of the electrophysiology and the second to imaging techniques.

Faculty

Stepahne Dieudonné, Ph.D. Institut de Biologie, CNRS UMR 8197 INSERM U 1024. Ecole Normale Supérieure, Paris, France.
Boris Barbour, Ph.D. Neuroscience Section, IBEns (CNRS UMR 8197 / INSERM U 1024) Ecole Normale Supérieure, Paris, France.
Mariano Casado, Ph.D. Neuroscience Section, IBEns (CNRS UMR 8197 / INSERM U 1024) Ecole Normale Supérieure, Paris, France.
Marco Diana, Ph.D. Institut de Biologie. CNRS UMR 8197 – INSERM U 1024.Ecole Normale Supérieure, Paris, France.
David Ogden, Ph.D. Laboratoire de Physiologie cérébrale, UMR8118, Université Paris Descartes, Paris, France.
Gabor Tamas, Ph.D. Department of Physiology, Anatomy and Neuroscience, University of Szeged, Hungary.

Organizers

Germán Szapiro, Ph.D. Neuroscience Section, IBEns (CNRS UMR 8197 / INSERM U 1024) Ecole Normale Supérieure, Paris, France.
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Juan Goutman, Ph.D. INGEBI-CONICET, Buenos Aires, Argentina.
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Outline Program

Day 1 - October 18th - Electrophysiology

8.00: Registration
09:00 - 11:00: 1st lecture. Membrane potential, action potential, excitatory and inhibitory synaptic currents (Mariano Casado).
This first lecture is intended to level the knowledge on basic neurophysiology among participants of the course.
11:05 - 11:20: Coffee brake.
11:30 - 12:55: 2nd lecture. Extracellular and intracellular solutions, basic properties, ions (David Ogden).
A discussion will be provided on the most common solutions used in cellular physiology, the basis for choosing specific ions, and also the underlying cellular principles.
13.00 - 14:00: Lunch.
14:00 – 18:00: 3rd lecture. Patch clamp techniques: voltage clamp, current clamp, perforated patch. (Gabor Tamas + Mariano Casado + Marco Diana).
The instructors will present a comparative analysis of different electrophysiological techniques, its applications and limitations. Typical problems, such as series resistance in patch-clamp recordings (whole cell configuration) will be evaluated. Practical approach. The choice of the appropriate techniques for each experimental condition and according to the parameter that needs to be measured.
18:00 - 18:30: Coffee brake.
18:30 - 20:30: 4th lecture.Electronics, the construction of a (basic) amplifier and test with model cell (Boris Barbour and David Ogden).
Practical approach. An introduction will be provided on the electronics used in patch-clamp, voltage-clamp amplifiers with microelectrods, current-clamp, etc. Students will be divided in small groups for discussion of details in their design of basic models of amplifiers. Demonstrations. Tests using oscilloscope and model cell.
20:45 - 21:15: Discussion. Round table.
A final session will be held among students and faculty in order to answer remaining questions related to the subjects of this first part of the course.
21.30 - 22:30: Dinner.

Day 2 - October 19th - Imaging

08:15 - 09:15 Breakfast Meeting
A breakfast will be served to students and professors before the first lecture. This seeks to give students the opportunity to have informal chats with the professors in order to discuss about their interests, future plans, career.
09:15 - 11:15. 5th lecture. Introduction to conventional and confocal microscopy (Stephanee Dieudonné).
This part is intended to level basic knowledge of students about the basis of optics and the principles and application of conventional and confocal microscopy. Concepts such as light source, laser, lamps, detectors will be discussed. As for the first lecture in the first section, this talk is meant to provide the students with the tools to understand the rest of the section.
11:15- 11:30: Coffee brake.
11:30 - 13:30: 6th lecture. Introduction to 2-photon microscopy (Stephanee Dieudonné).
The basis and applications of two photon microscopy will be explained. A comparison will be drawn with confocal microscopy.
13.45 - 14:30: Lunch
15:00 - 17:00: 7th lecture. Caged compunds and Ch- Rhodopsins. Principles, compounds (David Ogden and Marco Diana).
Ca2+ and glutamate uncaging as well as the novel Ch-Rhodopsin/allo-rhodopsin tools will be studied.
17:30 - 18:45: 8th lecture. Voltage sensitive dyes: principles – compounds – applications (David Ogden).
19:10 – 20:30: 9th lecture. Data analysis, programs (Boris Barbour).
A free software, WinWCP, for data acquisition will be presented. Examples from previous sections will be used to explain the principles of signal analysis in electrophysiology (i.e., spike sorting – cross correlogram – statistics). An open source -free - software for data analysis developed in Barbour’s lab will be introduced.
20:45 – 21:30: Round Table.
Final discussion on the contribution, perspectives and challenges of imaging techniques on neurophysiology. Remaining questions will be taken.
21.45 - 22:45: Dinner.
22.45: Social Event.