1. PETER PAZMANY CATHOLIC UNIVERSITY
SEMMELWEIS
UNIVERSITY
Development of Complex Curricula for Molecular Bionics and Infobionics Programs within a consortial* framework**
Consortium leader
PETER PAZMANY CATHOLIC UNIVERSITY
Consortium members
SEMMELWEIS UNIVERSITY, DIALOG CAMPUS PUBLISHER
The Project has been realised with the support of the European Union and has been co-financed by the European Social Fund ***
**Molekuláris bionika és Infobionika Szakok tananyagának komplex fejlesztése konzorciumi keretben
***A projekt az Európai Unió támogatásával, az Európai Szociális Alap társfinanszírozásával valósul meg.  
TÁMOP – /2/A/KMR

2. BASICS OF NEUROBIOLOGY - Methods
Peter Pazmany Catholic University
Faculty of Information Technology
Neurobiológia alapjai - Módszerek
BASICS OF NEUROBIOLOGY - Methods
By Imre Kalló
TÁMOP – /2/A/KMR

3. Basics of neurobiology: Methods in Neurobiology VII.
METHODS IN NEUROBIOLOGY VII.
Electrophysiological approaches
Imre Kalló Pázmány Péter Catholic University, Faculty of Information Technology
Histology techniques: light microscopic studies
Applications using fluorescent dyes
Histology techniques: electron microscopic studies
Techniques to map neuronal connections
Molecular biological techniques
Living experimental models
Electrophysiological approaches
Behavioral studies
Dissection, virtual dissection, imaging techniques
TÁMOP – /2/A/KMR
TÁMOP – /2/A/KMR

4. Basics of neurobiology: Methods in Neurobiology VII.
ELECTROPHYSIOLOGICAL APPROACHES
Electrophysiology is the study of electrical properties of biologycal cells and tissues.
In vivo electrophysiology
Anesthetised and moving animals
Measuring field potentials
Multiunit recordings
Single cell recordings
Juxtacellular recording
Intracellular recording
In vitro electrophysiology
Patch-clamp recording
Voltage clamp, Current clamp modes
Loose patch, Cell-attached, Whole cell, Perforated patch modes
TÁMOP – /2/A/KMR

5. Basics of neurobiology: Methods in Neurobiology VII.
MEASURING FIELD POTENTIALS
The field potential is the summation of spatial and temporal alterations of synaptic and voltage-dependent currents in a defined region of the brain. Consequently, it refers to and characterizes the activity of a certain cell or afferent population.
From the surface of the skull (EEG)
From the surface of the brain (ECoG)
From the brain (LFP, MUA) V
TÁMOP – /2/A/KMR

6. Basics of neurobiology: Methods in Neurobiology VII.
MEASURING FIELD POTENTIALS
Electroencephalography:
Neural oscillations:
δ oscillation: < 3.5 Hz θ oscillation: 4 – 7.5 Hz α oscillation: 8 – 13 Hz (Berger rythm) β oscillation: 14 – 30 Hz γ oscillation: > 30 Hz slow oscillations: < 1 Hz
From the surface of the skull (EEG)
From the surface of the rain (ECoG)
From the brain (LFP, MUA)
TÁMOP – /2/A/KMR

7. Basics of neurobiology: Methods in Neurobiology VII.
MEASURING FIELD POTENTIALS
From the surface of the skull (EEG)
From the surface of the rain (ECoG)
From the brain (LFP, MUA)
Tetrodes allow to visualisation of single neurons from different positions
Spike sorting, clustering
Local Field Potential (generated by membrane currents) input of the cells Multi Unit Activity (spiking of local neurons) output of the cells
TÁMOP – /2/A/KMR

8. Basics of neurobiology: Methods in Neurobiology VII.
IN VIVO EXTRACELLULAR RECORDING
TÁMOP – /2/A/KMR

9. Basics of neurobiology: Methods in Neurobiology VII.
IN VIVO EXTRACELLULAR RECORDING
MULTIELECTRODES
TÁMOP – /2/A/KMR

10. Basics of neurobiology: Methods in Neurobiology I.
IN VIVO JUXTACELLULAR RECORDING
An extracellular recording technique
It makes a correlated electrophysiological and morphological examination possible
Electrode and marker of cell filling (neurobiotin)
Fine positioning of the electrode – changes of the amplitude of action potentials
Current impulse (1-10 nA, anode impulse) – modulation of neuronal activitity
Duration of filling: min
TÁMOP – /2/A/KMR

11. Basics of neurobiology: Methods in Neurobiology VII.
IN VITRO ELECTROPHYSIOLOGY
VOLTAGE CLAMP
The cell’s potential is clamped at a chosen value. The size of ionic current crossing the cell’s membrane at the chosen value is measured.
Current meter
Membrane potential amplifier
Signal generator
Feedback amplifier
Intracellular electrode
Extracellular electrode
AXON
Circuit diagram depicting the feed-back amplifier charging the cell membrane of the recorded cell with the digitally subtracted transient capacitive currents.
Xu C et al Histamine innervation and activation of septohippocampal GABAergic neurones: involvement of local ACh release. J Physiol Dec 15;561(Pt 3):
TÁMOP – /2/A/KMR

12. Basics of neurobiology: Methods in Neurobiology I.
IN VITRO ELECTROPHYSIOLOGY
CURRENT CLAMP MODE
The current clamp technique records the membrane potential  by injecting current into a cell through the recording electrode. The membrane potential generated by the cell „spontaneously” or in response to external stimulus is measured.
Xu C et al Histamine innervation and activation of septohippocampal GABAergic neurones: involvement of local ACh release. J Physiol Dec 15;561(Pt 3):
TÁMOP – /2/A/KMR

13. Basics of neurobiology: Methods in Neurobiology I.
IN VITRO ELECTROPHYSIOLOGY: PATCH CLAMP TECHNIQUE
Cell-attached mode:
Transfer is only through ion channels entrapped
Whole cell mode:
Content of the pipette redistribute with that of the cytoplasm
Perforated patch mode:
Ion transfer through the perforations, but large molecules stay within the cytoplasm
TÁMOP – /2/A/KMR

14. Basics of neurobiology: Methods in Neurobiology VII.
IN VITRO RECORDINGS – PAIRED RECORDINGS
TÁMOP – /2/A/KMR