Electroencephalography

In this experiment, you will examine an electroencephalogram (EEG) and explore the electrical activity of the
brain. You will record an electroencephalogram from a volunteer, look at interfering signals, and examine the
effects of visual activity on alpha brain waves.

Ing. Fis. Juan Carlos Lizarazo MSc. (Modified from document written by staff of ADInstruments)

Background
The cerebral cortex contains huge numbers of neurons. Activity of these neurons is to some extent synchronized
in regular firing rhythms. These are referred to as brain waves. Electrodes placed in pairs on the scalp can pick
up variations in electrical potential that derive from this underlying cortical activity. The recording of the
electrical activity is called an electroencephalogram (EEG). EEG signals are affected by the state of arousal of the
cerebral cortex and show characteristic changes in different stages of sleep. EEG signals are also affected by
stimulation from the external environment and brain waves can become entrained to external stimuli.
Electroencephalography is used, among other things, in the diagnosis of epilepsy and the diagnosis of brain
death.

Recording the EEG

EEG recording is technically difficult, mainly because of the small size of the voltage signals, which are typically
50 µV peak-to-peak. The signals are small because the recording electrodes are separated from the brain's
surface by the scalp, the skull, and a layer of cerebrospinal fluid. A specially designed amplifier, such as the Bio
Amp built into the PowerLab, is essential to record EEGs. It is also important to use electrodes made of the right
material and to connect them properly. Even with these precautions, recordings may be spoiled by a range of
unwanted interfering influences, known as artifacts.

In this laboratory you will record EEG activity with two electrodes: a frontal electrode on the forehead, and an
occipital electrode on the scalp at the back of the head. A third (ground or earth) electrode is also attached, to
reduce electrical interference. In clinical EEG, it is usual to record many channels of activity from multiple
recording electrodes placed in an array over the head.

Figure 1. Equipment setup (with PowerLab 26T).

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Origins of the EEG signals

The EEG results from slow changes in the membrane potentials of cortical neurons, especially the excitatory and
inhibitory post-synaptic potentials (EPSPs and IPSPs). Very little contribution normally comes from action
potentials propagated along nerve axons. As with the ECG, the EEG reflects the algebraic sum of the electrical
potential changes occurring from large populations of cells. Therefore, large amplitude waves require the
synchronous activity of a large number of neurons. The rhythmic events that these waves reflect often arise in
the thalamus whose activity is in turn affected by a variety of inputs including structures in the brainstem
reticular formation.

Components of the EEG waveform

The EEG waveform contains component waves of different frequencies. These can be extracted and provide
information about different brain activities. The types of brain waves are:

❖ alpha (between 8 to 13 Hz; average amplitudes of 30 to 50 µV peak-to-peak) which will be studied in this
experiment. Alpha rhythm is seen when the eyes are closed and the volunteer relaxed. It is abolished
by eye opening and by mental effort such as doing calculations or concentrating on an idea. It is thus
thought to indicate the degree of cortical activation. The greater the activation, the lower the alpha
activity. Alpha waves are strongest over the occipital (back of the head) cortex and also over the frontal
cortex.
❖ beta (13 to 30 Hz; <20 µV peak-to-peak) which are prominent in alert individuals with their eyes open.
The beta rhythm may be absent or reduced in areas of cortical damage and can be accentuated by
sedative-hypnotic drugs such as benzodiazepines and barbiturates.
❖ theta (4 and 8 Hz; <30 µV peak-to-peak) which are seen in awake children but not adults. The theta
rhythm is normal during sleep at all ages. However, some researchers separate this frequency band into
two components, low theta (4 - 5.45 Hz) activity that they correlate with decreased arousal and
increased drowsiness, and high theta (6 - 7.45 Hz) activity that it is claimed is enhanced during tasks
involving working memory.
❖ delta (0.5 and 4 Hz; up to 100 - 200 µV peak-to-peak) which is the dominant rhythm in sleep stages three
and four but not seen in conscious adults. The delta rhythm tends to have the highest amplitude of any
of the component EEG waves. EEG artifacts caused by movements of jaw and neck muscles can produce
waves in the same frequency band.
❖ gamma (30 and 50 Hz). Most people recognize gamma rhythm, but its importance is controversial. It
may be associated with higher mental activity, including perception and consciousness and it
disappears under general anesthesia. One suggestion is that the gamma rhythm reflects the mental
activity involved in integrating various aspects of an object (color, shape, movement, etc) to form a
coherent picture. Interestingly, recent research has shown that gamma waves are enhanced in
Buddhist monks during meditation and are absent in schizophrenics.

It is not presently possible to relate the EEG waves to specific underlying neuronal activities. In general, the
more active the brain the higher the frequency and the lower the amplitude of the EEG. Conversely, the more
inactive the brain the lower the frequency and the higher the amplitude of the signal.

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The EEG during sleep

It is established that the EEG pattern provides an indicator of the sleep state. Sleep consists of two very different
alternating stages, non-REM and REM (rapid eye movements) sleep. Non-REM sleep is often described in four
stages that are characterized by a progressive increase in sensory thresholds, an increase in EEG wave
amplitude, and a decrease in EEG wave frequency. Stage 1 is marked by drowsiness and drifting in and out of
consciousness, this is followed by stages 2 and 3 and then 4. Sleepers then move back through the stages except
that rather than stage 1, REM sleep occurs. The whole cycle lasts approximately 90 minutes so that, over the
course of an 8 hour 'sleep', the cycle is repeated 4 to 6 times. In the later cycles, the REM component is longer
and stages 3 and 4 become shorter.

Figure 2. Sleep cycles.

These stages can be correlated with EEG activity. Stage 1 is associated with decreasing beta activity, alpha
activity that becomes less obvious and the emergence of theta activity. Stage 2 has irregular theta activity, short
bursts of waves of 12 - 14 Hz called sleep spindles, and sudden increases in wave amplitude (K complexes).

Figure 3. Sleep spindles.

Stages 1 and 2 are relatively "light" stages of sleep. In stages 3 and 4, delta activity predominates with the
distinction between the two being that in Stage 3 sleep there is delta activity for less that 50% of the time. In
stages 3 and 4 we are in deep sleep. In REM sleep, which can last from 20 to 60 minutes or more, the EEG is

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similar to that in Stage 1. REM sleep is the stage most associated with dreaming. Although the EEG shows
significant activity during REM sleep, motor activity is inhibited. Levels of brain serotonin and nor-epinephrine
alter during these sleep stages. In non-REM sleep stages 1 to 4, serotonin levels are increased whereas during

REM sleep, nor-epinephrine, corticosteroids and, in males, testosterone is secreted. Non-REM sleep is
characterized by decreases in blood pressure, and heart and respiratory rates. In REM sleep, there is marked
variation in heart rate and blood pressure and irregular breathing.

In sleep studies, EOGs and EMGs are often recorded in addition to the EEG. Non-REM sleep is characterized by
rolling, uncoordinated and slow eye movements and passively decreased muscle tone, whereas REM sleep has
rapid, coordinated eye movements (hence the name) and a little EMG activity reflecting the active inhibition of
muscle in this state.

Figure 4. Sleep stages.

The EEG and changes in intracranial metabolism

Changes in the EEG can be detected in response to changes in the chemical environment of the neurons. One
easy way to demonstrate this in a student laboratory is to observe the effects of hyperventilation.
Hyperventilation lowers blood PCO2. Since CO2, being lipid soluble, readily crosses the blood-brain barrier and
cell membranes, this in turn results in decreased PCO2 (hypocapnia) in the brain interstitial fluid and within the
neurons and glial cells. Thus, extracellular and cellular pH is elevated - acute respiratory alkalosis. In addition,
blood vessels in the brain constrict with reduction in brain blood flow. The consequences are a change in
neuronal activity with slower rhythms and higher amplitudes (increased delta and theta activities) as well as
some decrease in alpha activity. There is still debate about whether these EEG changes are a consequence of the
metabolic changes or of hemodynamic factors. One possibility is that they arise from depressant effects of the
hypocapnia on the brainstem reticular formation and are analogous to the EEG changes seen in the transition
from wakefulness to sleep.

The EEG and the functions of the cerebral hemispheres

Efforts have also been made to use EEG recordings to dissect out the contributions of the two hemispheres to
brain function. It has been argued that the left hemisphere is the 'logical' half of the brain concerned with
reasoning, problem solving and language while the right hemisphere is the more intuitive, creative side
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concerned with images and spatial processing rather than with language. Careful reading of the literature
reveals this to be a major oversimplification of cortical organization. In reality, there is little published EEG
evidence to lend credence to this hypothesis.

The EEG and personality

Attempts have also been made to relate personality to EEG patterns, perhaps the most famous example being
Eysenck's Cortical Arousal Model of Introversion and Extraversion. Eysenck argued that there is some 'optimal'
level of electrical activity in the cortex. If we fall below this we tend to be bored and fall asleep; above this we are
unable to deal with the activity and feel overwhelmed. In this construct, extraverts need additional mental
stimulation (people around them, loud music, etc) to reach this optimal cortical activity whereas introverts
avoid such additional stimulation as their cortical activity is already in the optimal region. There has been
considerable debate about the extent to which EEG findings support this hypothesis.

Further Reading

Kraemer et al., Nature, Vol. 434, Page 158 (2005).

What you will do in the laboratory

There are four exercises that you will complete during this Lab.

1. EEG artifacts. In this exercise, you will learn to recognize common artifacts seen while recording an EEG.

2. Alpha & Beta Rhythm. Here you will learn how best to elicit alpha waves in an EEG recording.

3. Effects of mental activity. In this part of the laboratory, you will do some simple arithmetic and observe the
effects on the EEG activity.

4. Effects of auditory stimulation. Here you will examine the effects on the EEG of the volume and the type of
music.

Required Equipment
o LabChart software
o PowerLab Data Acquisition Unit
o 5 Lead Shielded Bio Amp Cable
o EEG Flat Electrodes
o Electrode Paste
o Abrasive Gel or Abrasive Pad
o Alcohol Swabs
o Ballpoint pen
o Medical tape
o Elastic bandage

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Procedure
Equipment Setup and Electrode Attachment
1. Make sure the PowerLab is turned off and the USB cable is connected to the computer.

2. Connect the 5 Lead Shielded Bio Amp Cable to the Bio Amp Connector on the front panel of the PowerLab
(Figure 5). The hardware needs to be connected before you open the settings file.

Figure 5. Equipment Setup for PowerLab 26T

3. Attach the leads of the EEG Flat Electrodes to the Earth, CH1 NEG and POS pins closest to the labeled side on
the Bio Amp Cable. Channel 1 “positive” will lead to the inion (the bump on the back of the head above the
neck) and Channel 1 “negative” will lead to the forehead. Channel 2 will be empty and the Earth will lead to
the temple. Refer to Figure 5 for proper placement, but do not attach them to the volunteer. Follow the
color scheme on the Bio Amp Cable.

4. Remove any jewelry from the volunteer’s face, ears, and neck. Use a ballpoint pen to mark a small cross on
the skin on the back of the head, forehead, and temple. Use Figure 5 as a guide. Abrade the skin with
Abrasive Gel or Pad. This is important as abrasion helps reduce the skin’s resistance.

5. After abrasion, clean the area with an alcohol swab to remove the dead skin cells. While the skin is drying,
scoop Electrode Paste into the EEG Flat Electrodes. When the skin is dry stick the electrodes to the skin
(Figure 5). Immediately hold the electrodes and wires in place with the medical tape. Make sure the tape
firmly holds the electrodes against the head. Use the elastic bandage to wrap tightly around the head. This
will help the electrodes maintain good contact with the skin.

6. Have the volunteer lie in a comfortable position on his/her back, with the head turned so that none of the
electrodes are disturbed or compressed.

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7. Check that all three electrodes are properly connected to the volunteer and the Bio Amp Cable before
proceeding. Turn on the PowerLab.

Exercise 1: Recognizing Artifacts (artifacts file)

In this exercise, you will learn to recognize some of the artifacts that can appear on an EEG.

1. Launch LabChart and open the settings file “EEG Settings” from the Experiments tab in the Welcome
Center. It will be located in the folder for this experiment.

2. Select Bio Amp from the EEG Channel Function pop-up menu. It should be the only channel visible. Make
sure the settings are as follows: Range 200 µV, High Pass 0.5 Hz, and Low Pass 50 Hz.

Figure 6. Bio Amp Dialog

3. Start recording. Add a comment “blinking,” and have the volunteer blink repeatedly. Stop recording after
10 seconds.

4. Repeat step 3, this time, have the volunteer make eye movements. Add a comment “eye movements.”
Have the volunteer gaze up-and-down and left-and-right in a repeated pattern. Make sure the volunteer’s
head is still and only the eyes move.

5. Repeat step 3, this time, have the volunteer make head movements. Add a comment “head movements.”
Have the volunteer gently move his/her head in a repeated pattern.

6. Save your data, and open a new file with the same settings.

Analysis
Exercise 1: Recognizing Artifacts (artifacts file)
1. Examine the vertical scale at the left of the Chart View, and note the positions corresponding to +50 µV and –
50 µV. True EEG signals rarely exceed these limits.

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2. Examine the entire data trace and Autoscale, if necessary. There may be some large signals outside the ±75
µV range. Such large signals are artifacts.

Exercise 2: Alpha Waves in the EEG (shut open eye file)

In this exercise, you will examine the effects of relaxation and eye movement on alpha waves in the EEG.

1. Make sure the volunteer is relaxed and comfortable. Have the volunteer close his/her eyes and remain
quiet. Keep noise to a minimum and keep all distractions away from the volunteer.

2. Start recording. Record for 30 seconds. Prepare a comment with “open;” do not enter it yet.

3. Tell the volunteer to open both eyes. Immediately press Return/Enter to add the comment. Record with
the volunteer’s eyes open for 10 seconds. Do not stop recording.

4. Prepare a comment with “shut.” When the 10 seconds are complete, tell the volunteer to close both eyes.
Immediately press Return/Enter to add the comment.

5. Repeat steps 3 and 4 twice, to give you three sets of results. Save your data.

Analysis
Exercise 2: Alpha Waves in the EEG (shut open eye file)
1. Examine the entire data trace. Use the View Buttons to change the horizontal compression to see data with
eyes open and shut. Make a data selection that includes some data from both eyes open and eyes shut
conditions. View this selection in Zoom View. This should make it easier to see the alpha wave activity
(Figure 7). Now Autoscale, if necessary.

Figure 7. EEG signal with eyes shut and open. Note alpha waves during eyes shut.
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2. In Chart View scroll through the parts of the recording that were made with the volunteer’s eyes shut to look
for alpha waves. Use the View Buttons to change the horizontal compression if necessary. The alpha waves
can be recognized by their amplitude (usually 30 to 50 µV peak-to-peak, although it can be quite variable)
and their frequency. Each cycle of an alpha wave should last approximately 0.1 s.

Note: If you cannot find brain activity that matches the alpha rhythm, re-attach the electrodes and repeat this
exercise.

3. Use the Marker and Waveform Cursor to measure the amplitude of the alpha waves. Place the Marker at
the lowest point of the wave and move the Waveform Cursor to the peak of the wave. Measure the
amplitudes of five waves from when the volunteer’s eyes were closed. Record the values in Table 1 of the
Data Notebook of this document.

4. Now measure wave amplitudes when the volunteer’s eyes were open. Record these values in Table 2 of the
Data Notebook.

5. The amplitude of the waves is affected by the quality of the EEG signal. Therefore, it is useful to examine the
frequency and power of the wave activity. Spectral analysis can be used to examine these features of a
signal. Before examining your EEG signal you will complete a short tutorial on Spectral Analysis.

Spectral Analysis Tutorial

A spectrum is a representation of data based on the frequency distribution of its component sine waves. Spectra
indicate the strength of the various frequencies in a time-varying waveform. Spectrum View allows you to
observe the frequency distribution of data that might not otherwise be easily seen. For example, it could be used
to break down an EEG waveform into its various components: beta waves, alpha waves, theta waves and delta
waves. A mathematical technique known as the Fast Fourier Transform is applied to the raw data. The results of
this analysis can be displayed as a plot of the power (vertical axis) of different frequencies (horizontal axis)
relative to each other in the input signal. This is called a Power Spectrum Density (PSD) plot. The data can also
be displayed as 3-dimensional color plot of spectral power, frequency, and time called a Spectrogram.

1. Click on the home button on the lower left of the screen to open the Welcome Center. In the Experiments
tab browse the “EEG Spectral Analysis Tutorial.” It will be in the Settings folder for this experiment. Open
this file.

2. Examine the Chart View. Use the View Buttons to view each block. You should see five blocks of data. The
first record is a slowly oscillating sine wave

3. Open Spectrum view by clicking on the Spectrum View button in the Toolbar (Figure 4).

Figure 8. Spectrum View Toolbar button

4. Click the Smart Tile button in the LabChart Toolbar to display both windows in full screen mode.

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5. In Chart View Select the first record by double clicking in the Time axis. This will perform a spectral analysis
for this record and displays the result in the Spectrum view. Adjust the horizontal scaling of plots to view the
results:

➢ Set the horizontal scaling for the Power Spectrum Density (PSD) plot to 50 Hz (Figure 9). Use the
horizontal scroll bar to display the 0 Hz to 50 Hz region of the plot.

Figure 9. Spectrum PSD Scale

➢ Set the horizontal scaling for the Spectrogram to 50:1.

6. Examine the PSD plot and then the first section of the Spectrogram. Expand the vertical axes if necessary.
Use the waveform cursor to identify the frequency in Hertz (Hz) of the peak in the PSD plot and the band in
the Spectrogram. Values are displayed at the top of each plot.

➢ What is the frequency in Hertz (Hz) of this sine wave?

7. Select the second record and again view the result in the Spectrum view.

➢ What is the frequency in Hertz (Hz) of this second sine wave?

8. Select the third record and again view the result in the Spectrum view. You should now see two prominent
peaks (PSD plot) and bands (Spectrogram) in the result.

➢ Are these two peaks/bands the same as for the first two records?

9. Select the fourth record and again view the result in the Spectrum view.

➢ Is there any regular signal within this record?

10. In Chart View compare the signal amplitudes of the fourth and fifth records. Note that the fifth record has
lower amplitude compared with the fourth record.

11. Select the fourth record again. In the Spectrum view examine the PSD plot. Move the Waveform Cursor to
the prominent peak.

➢ What is the frequency (Hz) of this signal?

➢ What is the power (mV2) of this signal?

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12. Select the fifth record and examine the PSD plot. Move the Waveform Cursor to the peak.

➢ What is the frequency (Hz) of this signal?

➢ What is the power (mV2) of this signal?

13. Examine the Spectrogram. Note that the band corresponding to the signal’s frequency appears to missing.
This is because the power of the signal is small compared with the previous four records. Expand the scale
on the right hand side of the Spectrogram by setting the scale closer to the power of the signal (e.g., 0 to
0.004 V2) (Figure 6). Note that the band is now visible at the expected frequency.

Figure 10. Spectrogram Drop-down Menu and Set Scale Dialog

14. The fifth record is the same signal as the fourth record, except that the quality of the raw signal has been
affected. Compare your features (amplitude, power, frequency) of the fourth and fifth record.

➢ How has the quality of the signal affected the wave features?

Exercise 2: Alpha Waves in the EEG contd.

Now you will use Spectral Analysis to examine the EEG you recorded.

1. Use the View Buttons to change the horizontal compression to 10:1 (Figure 11).

Figure 11. Alpha Waves with 10:1 Compression

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2. Find the part of the recording when the volunteer had his/her eyes shut. Click-and-drag across this part of
the data trace to select it. From the Window menu, select Spectrum. In the Spectrum View choose
Selected (Figure 12).

Figure 12. Spectrum of an EEG

3. Alpha activity shows up in the PSD plot as a clear peak in the 8-12 Hz range. Then print the PSD plot.

4. Alpha activity shows up in the Spectrogram as a band of color in the 8-12 Hz range. If you cannot see the
alpha activity as a clear peak in the 8-12 Hz range, scale the horizontal and vertical axes. Note that
Spectrogram displays all the recorded data and that the selection you have made is highlighted in a darker
blue color.

5. Make a data selection of several seconds from when the volunteer had their eyes open. Select Spectrum.
Note that in the PSD plot the peak in the alpha activity range of 8-12 Hz is small or absent and in the
Spectrogram the band of color in the alpha activity range of 8-12 Hz is weak or absent. Print the PSD plot.

6. In the Spectrogram scale the horizontal axis so that all the data is visible. Note the presence and absence of
the band of color in the alpha activity range of 8-12 Hz which correspond with the eyes shut and eyes open
conditions. Print the Spectrogram.

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Data Notebook
Table 1. Alpha Waves
Amplitude
Wave #1

Wave #2

Wave #3

Wave #4

Wave #5

Table 2. Absence of Alpha Waves

Amplitude
Wave #1

Wave #2

Wave #3

Wave #4

Wave #5

Exercise 3: Effects of Mental Activity (arithmetic file)

1. Select a registry region

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2. Select the Data Pad icon indicated in the following figure.

3. With the data from the Data Pad, proceed to fill in the corresponding tables.

Analysis
Exercise 3: Effects of Mental Activity (arithmetic file)

Alpha Waves

Condition Average Amplitude (µV) Average Frequency (Hz)

No arithmetic

No arithmetic

No arithmetic

Media

DE

Arithmetic

Arithmetic

Arithmetic

Mean

DE

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Beta waves

Condition Average Amplitude (µV) Average Frequency (Hz)

No arithmetic

No arithmetic

No arithmetic

Mean

DE

Arithmetic

Arithmetic

Arithmetic

Mean

DE

Exercise 4: Effect of Auditory Stimulation (auditory file)

Repeat the procedure from exercise 3

Analysis
Exercise 4: Effect of Auditory Stimulation (auditory file)

Alpha Waves

Condition Average Amplitude (µV) Average Frequency (Hz)

No music

Classic, low

No music

Classic, high

No music

Rock, low

No music

Rock, high

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Beta waves

Condition Average Amplitude (µV) Average Frequency (Hz)

No music

Classic, low

No music

Classic, high

No music

Rock, low

No music

Rock, high

Study Questions
1. How does the human anatomy make it difficult to record an electroencephalogram, or EEG?

2. What are common causes of the artifacts you recorded in Exercise 1? Name at least three causes.

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3. In Exercise 2, under what conditions did you see alpha waves more clearly?

4. What are alpha waves thought to indicate?

5. Look at your Spectral Analysis from when the volunteer had their eyes closed. At what range do you
have the greatest peak? What type of brain waves does the Spectral Analysis suggest is active?

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6. Look at your Spectral Analysis from when the volunteer had their eyes open. At what range do you
have the greatest peak? What type of brain waves does the Spectral Analysis suggest is active?

7. What was the effect of mental arithmetic on alpha wave activity?

8. How do you explain your findings?

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9. What effects did different types of music have on alpha wave activity?

10. How do you explain your findings?

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