Lesson 1-2 Main Research Methods

Research Methods in
Cognitive Neuroscience
lab
Prof Alice M. Proverbio, PhD

 Zani A & Proverbio A.M. (2003) The Cognitive electrophysiology of mind and brain,
Academic Press/Elsevier, New York. Foreward by Michael Posner.
 Proverbio AM, Zani A. (2013). Metodi Strumentali nelle Neuroscienze Cognitive. EEG
ed ERP- Instrumental Methods in Cognitive Neuroscience. EEG and ERP. Edition: 1st,
Aracne Editrice: Roma. ISBN: 978-88-548-6655-3
 Psicofisiologia Cognitiva, Elettrofisiologia della mente, Carocci editore
Bicocca ERP lab https://bicoccaerplab.wordpress.com/ Room 3140 U6 3° floor

Head of Cognitive Electrophysiology lab: BicoccaErpLab
mado.proverbio@unimib.it
(Office hours: Monday 9:00-10:00)
Twitter @AliceProverbio
Lab attendance is required:
How to certify attendance:
1. By signing in presence - max 2 lessons can be missed)

2. By participating to the practical activities
CLASS timetable: Research Methods in Cognitive Neuroscience
07-03-2023 14:30 - 17:30 RES METH COGN NEUROSCI
31-03-2023 8:30-12:30 ?
Practical hands on activities:
We will learn:
A. How to choose the more appropriate neuroimaging techiques for addressing
a specific research problem
B. Fundamentals of EEG/ERP technique: How to correcly place EEG electrodes
and Electrocaps. How to find standard EEG markers on the skull bones.
C. How to quantify latency and amplitude of electric peaks
D. How to write a Clinical report on real EEG data from human patients
E. How to develop a novel ERP experimentalparadigm on P300 component (to
be designed by you)
• Analyses of real EEG traces of different types of patients. Each student

will be given 1-2 examples of Neurophysiopathological data, and will
have to write a short clinical report, either individually or in
cooperation with another colleague
• (Individual reading of the report and discussion in class)
• VISIT TO EEG/ERP lab in presence and hands on activity
1. Fundamentals in EEG/MEG, including the electrophysiological
bases of signals recorded in awake, sleep and pathological
states, instrumentation, and experimental designs.
2. Application of EEG/ERP technology in the study of the neural

bases of cognition (Cognitive Electrophysiology).
3. New advances in EEG/ERP applications (e.g., Forensic

Neuroscience and lies detection, Music Perception and Music
therapy, Artificial intelligence, Brain Computer Interface, “Mind
reading”, Relaxation Techniques for Stress Relief (e.g. Yoga),
Neurofeedback for ADHD treatment, Neuroaesthetics,
Neuroeconomics, Political Neuroscience, Sexual gender studies,
Racial prejudice, Consciousness detection in coma patients,
Appreciation and aesthetic evaluation of brands/products, etc..)
Cognitive Electrophysiology of Mind
and Brain presents the latest
developments in the study of the
relationships between mind and brain
as revealed using the recording of
bioelectric (ERPs) and magnetic
responses (ERFs) of the brain. To a
more limited degree, PET and fMRI
measures of brain activity are also
covered, in an effort to show how the
different signals of brain activation can
be combined, and how they can
complement each other in the
elucidation of brain distributed
systems involved in human cognition.
Emphasis is placed on reviewing
experimental findings in the areas of
attention, language, memory, visual
and auditory perception, and brain
damage research.
Time-lapse phase contrast microscopy of neuronal cells and
keratinocytes. Images were acquired on an Olympus IX3 inverted
microscope every 108 seconds over a period of 18hr using 10X
objective.
FUNCTIONAL NEAR-INFRARED SPECTROSCPY FNIRS
Optical imaging technique
Light is delivered to, and collected from the sample, by means of optical fibers
(optodes)
A weak (mW) light signal is injected in the tissue and the emitted signal which
carries information of tissue constituents is measured.
The difference in the absorption spectra of oxygenated and deoxygenated haemoglobin allows separate
measurements.
The interplay between light absorption and light scattering determines the overall light attenuation.
Detected light provides a measure of changes in cerebral haemoglobin concentrations, as revealed by

changes in optical absorption
Near-infrared light is able to propagate several
centimeters through tissue, through the scalp and skull,
and spectroscopically interrogate the concentrations of
oxygenated, de-oxygenated, and total hemoglobin within
the brain
Changes in the amount of diffuse light reaching the

detector corresponds to changes in the optical properties
of the tissue below and between the source and detector
Structural Brain Imaging Techniques
CAT
Different absorption coefficient of X rays (depending on tissue density)

The white and gray matter of the brain can be clearly differentiated by computed
tomography (CT). The differentiation is enhanced by the administration of contrast
material. Without contrast material, the mean attenuation number of the white matter is 29
units, of the gray matter, 35 units.
Structural Brain Imaging Techniques
MRI
MRI - grey matter: intermediate signal intensity (grey)

white matter: hyper-intense resonance signal compared to grey matter (white-ish)
On a T1-weighted scans show tissues with high fat content (such as white matter) appear
bright and compartments filled with water (CSF) appears dark.
Neurometabolic functional techniques
Positron
emission
tomography
The cylindrical tube of an MRI scanner houses a very
powerful electro-magnet. A typical research scanner has a field
strength of 3 teslas (T), about 50,000 times greater than the
Earth’s field.
The magnetic field inside the scanner affects the magnetic nuclei
of atoms.
Normally atomic nuclei are randomly oriented but under the
influence of a magnetic field the nuclei become aligned with the
direction of the field. The stronger the field the greater the
degree of alignment.
When pointing in the same direction, the tiny magnetic signals
from individual nuclei add up coherently resulting in a signal that
is large enough to measure. In fMRI it is the magnetic signal from
hydrogen nuclei in water (H 2O) that is detected.
The key to MRI is that the signal from hydrogen nuclei varies in
strength depending on the surroundings. This provides a means
of discriminating between grey matter, white matter and
cerebral spinal fluid in structural images of the brain.
Functional magnetic resonance
imaging (fMRI)
Oxygen is delivered to neurons by haemoglobin in capillary red blood cells. When

neuronal activity increases there is an increased demand for oxygen and the local
response is an increase in blood flow to regions of increased neural activity.
Haemoglobin is diamagnetic when oxygenated but paramagnetic when deoxygenated.
This difference in magnetic properties leads to small differences in the MR signal of blood
depending on the degree of oxygenation. Since blood oxygenation varies according to the
levels of neural activity these differences can be used to detect brain activity. This form of
MRI is known as blood oxygenation level dependent (BOLD) imaging.
Functional magnetic resonance
imaging (fMRI) (Part 1)
Functional magnetic resonance imaging (fMRI)
There is a momentary decrease in blood oxygenation immediately after neural
activity increases, known as the “initial dip” in the haemodynamic response. This is
followed by a period where the blood flow increases, not just to a level where
oxygen demand is met, but overcompensating for the increased demand.
This means the blood oxygenation actually increases following neural activation.
The blood flow peaks after around 6 seconds and then falls back to baseline, often
accompanied by a "post-stimulus undershoot".
Stimulus onset
Experimental designs in PET and fMRI
studies (Part 1)
Event-related design
Temporal resolution
Event-related fMRI responses
Intraoperative electrocorticography
Transcranial magnetic stimulation
(TMS)
TMS involves the delivery of a brief (~200 µs) and powerful (0.2 to 4.0 T) magnetic pulse
to the scalp through a coil. The magnetic pulse induces a transitory electric current in
the cortical surface under the coil, which causes the depolarization of cell membranes
and transynaptic depolarization of a population of cortical neurons.
Risoluzione spaziale = 1,0-1,5 cm3
Single impulse or repetitive

Repetitive:
1 Hz inhibitory
5 Hz excitatory
Common side effects
o Headache
o Scalp discomfort at the site of stimulation
o Tingling, spasms or twitching of facial muscles
o Lightheadedness
Uncommon side effects
o Seizures
o Mania, particularly in people with bipolar disorder
o Hearing loss if there is inadequate ear protection during treatment
Due to the strong magnetic field produced during rTMS, the procedure is
not recommended for people having:
Aneurysm clips or coils
Stents
Implanted stimulators
Implanted vagus nerve or deep brain stimulators
Implanted electrical devices, such as pacemakers or medication pumps
Electrodes for monitoring brain activity
Cochlear implants for hearing
Any magnetic implants
Bullet fragments
Transcranial Direct-Current Stimulation (tDCS)
Portable, non-invasive neuromodulatory technique that delivers a low
electric current to the scalp.
A fixed current between 1 and 2 mA is typically applied.
tDCS works by applying a positive (anodal) or negative (cathodal) current
via electrodes to an area, facilitating the depolarization or
hyperpolarization of neurons, respectively. The positioning of the anode
and cathode electrodes is used to influence how current flows, and where
in the brain it does.
To stimulate the left STG (anodal left/cathodal right stimulation), the anode electrode
was positioned centered over T3 site.
To stimulate the right STG (anodal right/cathodal left stimulation), the anodal electrode
was positioned centered over T4 site.
Anodal right/Cathodal left condition (rSTG
stimulation) significantly improved the
response times specifically for Comic stimuli
relative to the Sham condition.
Overall, these results suggest that the anodal

right/cathodal left STG condition improved
subjects’ ability to recognize the comic
element of the misfortunate situation. This
also suggests that facial expression recognition
is involved in humor processing.
Neurophysiology
An electrode introduced into the brain of a living animal will

detect electrical activity that is generated by the neurons
adjacent to the electrode tip. If the electrode is a
microelectrode, with a tip size of 3 to 10 micrometers, the
electrode will often isolate the activity of a single neuron.
1 micron, micrometre is one millionth of a metre (or one
thousandth of a millimeter)
Recording single-unit activity in an
awake, behaving monkey
In the winter of 1991 in Parma, Giacomo Rizzolatti, Giuseppe Di Pellegrino,
Luciano Fadiga, Leonardo Fogassi and Vittorio Gallese found mirror neurons in the
ventral premotor cortex of the monkey.
Electroencephalographic recording
(EEG)
Event-related potentials (ERPs)
Event-related potentials (ERPs)
Electrophysiological basis of EEG and
ERP signals
Contour maps showing spatial-
temporal patterns of ERP activity
Magnetoencephalography
Magnetoencephalographic signals
Magnetoencephalographic signals
Somatosensory potentials (fingers and toes)
Hundreds of lasers project light into the brain and measure
the intensity and delay of light coming out with photo-
detectors. This is called Diffuse optical imaging, and the
data are called the event-related optical signal.
Diffusion Tensor Imaging (or DTI), from diffusion tensor imaging).
Used to study the anatomical structure of the axon tracts that form
the brain white matter (connectivity between different regions)
Associations, dissociations, and double
dissociations
Spatial and temporal resolution of the
methods
Combining information across subjects localizes brain region underlying cognitive deficit

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Research Methods in

Prof Alice M. Proverbio, PhD

 Psicofisiologia Cognitiva, Elettrofisiologia della mente, Carocci editore

Bicocca ERP lab https://bicoccaerplab.wordpress.com/ Room 3140 U6 3° floor

How to certify attendance:

1. By signing in presence - max 2 lessons can be missed)

07-03-2023 14:30 - 17:30 RES METH COGN NEUROSCI

09-03-2023 14:30 - 17:30 RES METH COGN NEUROSCI

14-03-2023 14:30 - 17:30 RES METH COGN NEUROSCI

16-03-2023 14:30 - 17:30 RES METH COGN NEUROSCI

21-03-2023 14:30 - 17:30 RES METH COGN NEUROSCI

23-03-2023 14:30 - 17:30 RES METH COGN NEUROSCI

28-03-2023 14:30 - 17:30 RES METH COGN NEUROSCI

30-03-2023 14:30 - 18:30 RES METH COGN NEUROSCI

• Analyses of real EEG traces of different types of patients. Each student

2. Application of EEG/ERP technology in the study of the neural

3. New advances in EEG/ERP applications (e.g., Forensic

Detected light provides a measure of changes in cerebral haemoglobin concentrations, as revealed by

Changes in the amount of diffuse light reaching the

Different absorption coefficient of X rays (depending on tissue density)

MRI - grey matter: intermediate signal intensity (grey)

Oxygen is delivered to neurons by haemoglobin in capillary red blood cells. When

Single impulse or repetitive

Overall, these results suggest that the anodal

An electrode introduced into the brain of a living animal will

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