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 International Journal of Innovative Research in Advanced Engineering (IJIRAE) ISSN: 2349-2163 Volume 1 Issue 9 (October 2014 ) www.ijirae.com © 2014, IJIRAE- All Rights Reserved Page - 65 Independent Component Analysis of EEG Signals and Real Time Data Acquisition Using MyDAQ and Labview K.Vijayalaks hmi S. Ramachandran M.Chandras ekaran  Research Sc holar SJB InstituteT echnology Govt. Colleg e of Engine ering VMU, Salem Bangalore Bargur, Krishnagiri  Abstract - Electroencephalography is the recording of electrical activity along the scalp of a person.  This allows us to  measure brain activity involved in various t ypes of cognitive functions . Experimental goal of this work is to interpret and  characterize the EEG activity during Pranayama breathing with respect to temporal and spatial context, and acquire two  channel data using MyDAQ and Labview. This work primarily explores the variation in the EEG wave pattern during  different stages of Pranayama as well as the variation of alpha wave level i n the left and right frontal, temporal parietal and  occipital regions of the brain. Statistical significance test is performed for different cycles of Pranayama to measur e the  significant change in the alpha power wi th respect to baseline measures. An effort wa s made to analyze t he differenc e in the  cerebral electrical activity among long term and short term meditation practitioners. Data was recorded for ten subjects  during three cycles of Pranayama, e ach cycle lasting for two minutes. In order to measure t he effects towards the end of  Pranayama, the last 20 seconds of EEG data were analyzed in ea ch cycle of Pranayama. The analysis revealed that 40 % of  the subjects were relaxed (means increase in alpha power) as well as alert (means increase in beta power) at the end of  Pranayama, whereas 30% of the subjects showed decrease in beta power. Al so, 10% showed increase in beta power for only  one cycle. Therefore, one may conclude that the practice of Pranayama enhances relaxation and cognition, leading the  practitioners to a stress f ree and healthy lif e.   Keywords: Electroen cephalogram, Inde pendent Componen t Analysis, Artifacts, Data Ac quisition, Labview I. INTRODUCTION There is growing interest in the integration of meditation in clinical applications for treatment therapy and in the education field to have the benefits of meditation. This work presents the evidence related to the neuro physiological changes during meditation. Meditation facilitate to achieve educational goals, increases student mental calibre in spite of academic stress, and to enhance the very life style of the person. In short, meditation contributes sub stantially towards crafting the most eff ective cognitive training programs. The Pranayama is derived from two Sanskrit words ‘Prana’ and ‘ayama’, where ‘Prana’ means Energy or life and ‘ayama’ means elongation of life span. The word Pranayama, therefore, means elongation of pranic energy. There are three Stages of Pranayama used in the scriptures [1-3]. They are: 1 st  Stage: 8-10 times Ujjai breath (meaning deep), Posture: Vajrasana, Thumbs on the hip bone, Palms parallel to the ground, spine erect followed by 30-45 seconds Relaxation. This i s shown in Figure 1a. 2 nd  Stage: 8-10 times Ujjai breath (Posture: Vajras ana, Thumbs under arm pits, Palms para llel to the ground, spine erect) as shown in Figure 1b and followe d by 30-45 seconds Relaxation. 3 rd Stage: 8-10 times U jjai breath (Posture: Vaj rasana, palms on the back, bice ps touching the ear and elbow pointing to the ceiling, spine erect, chin at th e normal level), followed by 30-45 seconds Relaxation. This is shown in Figure 1c. During normal breathing, we use only half the lung capacity, which can be easily understood by taking a deep breath filling the entire lungs. During the practise of Pranayama, we use at least 80% of our lung capacity. Deeper the breath more is the oxygen entering the blood, thus benefitting every blood cell and consequently the whole body. The mind and the breath have a common source, which means our thoughts and breath are directly related. For instance, when we are angry or restless, the number of  breaths per minute also increases rapidly. By controlling the breathing, one can control unnecessary thoughts and also the emotions, aided substantially by the practise of Pranayama. In short, if breath i s controlled, then the thoughts are also cont rolled. . Fig. 1 Pranayama i n Vajrasana Posture (a) Fi rst Stage: Adhama, (b) Second St age: Madhyama (c) Third Stage: U ttama (a) First Stage (b) Second Stage (c) Third Stage

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Page 1: IJIRAE:: Independent Component Analysis of EEG Signals and Real Time Data Acquisition Using MyDAQ and Labview

8/10/2019 IJIRAE:: Independent Component Analysis of EEG Signals and Real Time Data Acquisition Using MyDAQ and Labview

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  International Journal of Innovative Research in Advanced Engineering (IJIRAE) ISSN: 2349-2163 

Volume 1 Issue 9 (October 2014)  www.ijirae.com 

_______________________________________________________________________________________________________

© 2014, IJIRAE- All Rights Reserved Page - 65

Independent Component Analysis of EEG Signals and

Real Time Data Acquisition Using MyDAQ and Labview 

K.Vijayalakshmi S. Ramachandran M.Chandrasekaran 

 Research Scholar SJB InstituteTechnology Govt. College of EngineeringVMU, Salem Bangalore Bargur, Krishnagiri

 Abstract - Electroencephalography is the recording of electrical activity along the scalp of a person.  This allows us to measure brain activity involved in various types of cognitive functions. Experimental goal of this work is to interpret and

 characterize the EEG activity during Pranayama breathing with respect to temporal and spatial context, and acquire two channel data using MyDAQ and Labview. This work primarily explores the variation in the EEG wave pattern during different stages of Pranayama as well as the variation of alpha wave level in the left and right frontal, temporal parietal and occipital regions of the brain. Statistical significance test is performed for different cycles of Pranayama to measure the significant change in the alpha power with respect to baseline measures. An effort was made to analyze the difference in the

 cerebral electrical activity among long term and short term meditation practitioners. Data was recorded for ten subjects during three cycles of Pranayama, each cycle lasting for two minutes. In order to measure the effects towards the end of Pranayama, the last 20 seconds of EEG data were analyzed in each cycle of Pranayama. The analysis revealed that 40 % of

 the subjects were relaxed (means increase in alpha power) as well as alert (means increase in beta power) at the end of Pranayama, whereas 30% of the subjects showed decrease in beta power. Also, 10% showed increase in beta power for only one cycle. Therefore, one may conclude that the practice of Pranayama enhances relaxation and cognition, leading the

 practitioners to a stress free and healthy life.  Keywords: Electroencephalogram, Independent Component Analysis, Artifacts, Data Acquisition, Labview

I. INTRODUCTIONThere is growing interest in the integration of meditation in clinical applications for treatment therapy and in the

education field to have the benefits of meditation. This work presents the evidence related to the neuro physiological changes

during meditation. Meditation facilitate to achieve educational goals, increases student mental calibre in spite of academic stress,and to enhance the very life style of the person. In short, meditation contributes substantially towards crafting the most effectivecognitive training programs.

The Pranayama is derived from two Sanskrit words ‘Prana’ and ‘ayama’, where ‘Prana’ means Energy or life and

‘ayama’ means elongation of life span. The word Pranayama, therefore, means elongation of pranic energy. There are threeStages of Pranayama used in the scriptures [1-3]. They are:

1st Stage: 8-10 times Ujjai breath (meaning deep), Posture: Vajrasana, Thumbs on the hip bone, Palms parallel to the ground,

spine erect followed by 30-45 seconds Relaxation. This is shown in Figure 1a.2

nd  Stage: 8-10 times Ujjai breath (Posture: Vajrasana, Thumbs under arm pits, Palms parallel to the ground, spine erect) as

shown in Figure 1b and followed by 30-45 seconds Relaxation.3rd Stage: 8-10 times Ujjai breath (Posture: Vajrasana, palms on the back, biceps touching the ear and elbow pointing to the

ceiling, spine erect, chin at the normal level), followed by 30-45 seconds Relaxation. This is shown in Figure 1c.

During normal breathing, we use only half the lung capacity, which can be easily understood by taking a deep breath filling the

entire lungs. During the practise of Pranayama, we use at least 80% of our lung capacity. Deeper the breath more is the oxygenentering the blood, thus benefitting every blood cell and consequently the whole body. The mind and the breath have a common

source, which means our thoughts and breath are directly related. For instance, when we are angry or restless, the number of breaths per minute also increases rapidly. By controlling the breathing, one can control unnecessary thoughts and also theemotions, aided substantially by the practise of Pranayama. In short, if breath is controlled, then the thoughts are also controlled.

.

Fig. 1 Pranayama in Vajrasana Posture (a) First Stage: Adhama, (b) Second Stage: Madhyama (c) Third Stage: Uttama

(a) First Stage (b) Second Stage (c) Third Stage

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  International Journal of Innovative Research in Advanced Engineering (IJIRAE) ISSN: 2349-2163 

Volume 1 Issue 9 (October 2014)  www.ijirae.com 

_______________________________________________________________________________________________________

© 2014, IJIRAE- All Rights Reserved Page - 66

An effort was made to analyze the cerebral electrical activity for ten subjects. EEG is recorded during premeditation as a

 baseline and three cycles of Pranayama. It is a slow and fast breathing exercise. This work is an attempt to score the effect ofPranayama by studying the variation in EEG voltage levels [4]. Most of the referred papers have not presented quantitativeanalysis [1-8]. Papers on Meditation referred earlier have performed either coherence or qualitative analysis and data has been

acquired only for 4 or 5 subjects

Therefore, aim of our work is to perform a detailed quantitative and qualitative analysis acquiring data for 10 subjects, offering better results. EEG signal comprises five low frequency bands, namely, Alpha (8-13 Hz), Beta (14-30 Hz), theta (4-8 Hz), Delta

(0.5-4 Hz) and Gamma (above 30 Hz). Our objective is to do a quantitative analysis of these bands during Pranayama. EEG datafor ten subjects was acquired using an RMS Recorder with 10-20 Electrode system. This data was converted into a 32 channeldata by setting to a vertical common Average Reference. The 32 Channel data is used to obtain a Topo plot for further data

analysis. 4th

 order Elliptic filter is applied for the acquired EEG data to extract EEG waves, namely Delta (below 3 Hz), Theta(4-7 Hz), Alpha (8-13 Hz), Beta (14 Hz and above). Through Fast Fourier Transform, power in each wave is calculated [5-8].

Independent Component Analysis (ICA) is a quite powerful higher order statistical technique and is able to separate independentsources linearly mixed in several sensors. For instance, when recording electroencephalograms (EEG) on the scalp, ICA canseparate out artefacts embedded in the data (since they are usually independent of each other). A first step in many ICAalgorithms is to whiten (or sphere) the data. This means that we remove any correlations in the data, i.e., the different channels

are forced to be uncorrelated.

The identification of independent components from EEG signal helps to understand better the sources of EEG signal.This was demonstrated by Hyvarinen A et al. [9]. Resolving of the sources of artefacts and the sources of useful signals are dealt

in his work. The use of Independent Component Analysis as a progressive method for removing artefact from EEG signal was presented by Bell A. et al. [10]. The work has helped in recovery of epileptic peak sources, spontaneous activity sources andvarious grapho elements as K-complexes and sleep spindles. The use of EEG signals to estimate cognitive state was proposed by

Boscolo R. et al. [11]. The estimation was done using a mutual information based method. Classification was done in their work by employing a group of three classifiers and using majority voting logic. However, the problem of finding robust features that

allows cross-session and cross-subject generalization are not discussed in their work. The combined use of ICA for pre- processing the EEG signal, Discrete Wavelet Transform analysis for feature extraction and fuzzy cluster means algorithm forrecognition of some diseases were proposed by Delorme A et al. [12].

Quantitative analysis of EEG signals is a challenge in the cognitive assessment or for diagnostic Applications. In this

work, ICA is implemented for artefact removal and for source localisation.This paper is organized as follows. In section two, the methodology of EEG processing is presented in detail. Section

three presents results. Finally, conclusion is drawn.

II. EEG ACQUISITION AND ANALYSIS

EEG signals were acquired for ten normal subjects between the age group of 25 to 40 years. Five of them are practicingPranayama for more than eight years, considered as long term Practitioners; otherwise taken as short term practitioners. EEG is

recorded according to International standard 10-20 Electrode system using Recorders Medicare Systems with 24 channel digitalEEG machine having an A/D conversion of 16 bits with sampling frequency of 256 Hz, Software version of Super Spec. 4.2.54.Here, 10-20 denotes the placement of

Fig. 2 Placement of International Standard 10-20 Electrode

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  International Journal of Innovative Research in Advanced Engineering (IJIRAE) ISSN: 2349-2163 

Volume 1 Issue 9 (October 2014)  www.ijirae.com 

_______________________________________________________________________________________________________

© 2014, IJIRAE- All Rights Reserved Page - 67

Fig. 3 Display of 18 Channel EEG Data X axis: No. of samples; Y axis: Amplitude in µvolts 

Fig. 4 Procedure for EEG Data Acquisition and Analysis

the electrodes in a particular manner as shown in Figure 2. EEG is recorded during Pre Meditation stage for first 5 minutes witheyes closed and followed by 3 cycles of Pranayama for six minutes. EEG data for last 20 seconds from each cycle of Pranayamais taken for analysis. EEG data acquired from a 10-20 system has 18 channels as shown in Figure 3; most of the channels having

redundant data. Principal Component Analysis (PCA) is applied on the acquired data for dimensionality reduction. By applyingPrincipal component analysis, we get “Scree” plot, from which it is found that 5 channels that have maximum EEG amplitude

variations are adequate for analysis as shown in Figure 4. But PCA does not identify which of the 18 channels have maximumEEG amplitude variations. The Independent Component Analysis is applied to find the Maximum EEG variation channels. Inthis work, Independent Component Analysis is implemented using EEG LAB open source software [13].

Fig.5 Scree Plot: Variance vs Channel in an EEG Data

Principal Component Analysis

EEG Data Selection

Independent Component Analysis

Extraction of Delta, Theta, Alpha and Beta Waves

Power Calculation of Delta, Theta, Alpha and Beta Waves

EEG Data Acquisition

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  International Journal of Innovative Research in Advanced Engineering (IJIRAE) ISSN: 2349-2163 

Volume 1 Issue 9 (October 2014)  www.ijirae.com 

_______________________________________________________________________________________________________

© 2014, IJIRAE- All Rights Reserved Page - 68

The goal of this work is to interpret and characterize the EEG activity during Pranayama with respect to temporal and spatialcontext. This work primarily analyses the variation in the EEG wave pattern during different cycles of Pranayama as well as

variation of alpha wave power in different regions of the brain. Recording Conditions were set for the subjects to be verycomfortable. Subjects meditated with their eyes closed in a sitting position in a room free of sound and visual distractions. EEGactivity is complex in nature. Therefore every effort has been made to minimize the artefacts that may affect analysis and

results. The Methodology implemented is to interpret the frequency waves such as Delta, Theta, Alpha, and Beta that

characterize the brain state. The analysis is based on these specific frequency ranges in order to interpret the changes thatoccurred during meditation.Independent Component Analysis: An electrode sensing a signal at a position may be influenced by the signals picked up byneighbouring electrodes. ICA is a quite powerful technique which is capable of obtaining the desired signals filtering out the

unwanted neighbouring signals. ICA can separate out artefacts embedded in the data since they are usually independent of eachother. A first step in ICA algorithm is to centre a data by subtracting each channel data from its mean value. This is referred to as“white” or “sphere”. Whitening the data is a Pre-processing step performed by most ICA algorithms before applying ICA. Thisstep removes any correlations in the data, i.e., all channels are forced to be uncorrelated. A geometrical interpretation for two

mixed random sinusoidal sources A and B with different amplitudes and frequencies is shown in Figure 6, where the two signalsare linearly mixed. This results in Gaussian distribution. If we whiten the two linear mixtures, we get the plot as in Figure 7. Thevariance on both axes is now equal and the correlation of the projection of the data on both axes is 0, meaning that the

covariance matrix is diagonal and that all the diagonal elements are equal. The whitening process is simply a linear change ofcoordinate of the mixed data. Once the ICA solution is found in this "whitened" coordinate frame, we can easily re project the

ICA solution back into the original coordinate frame.

Fig. 6 Two Mixed Random sources A vs B

Fig. 7 Whitening of the A and B Mixed Data

ICA components are arranged as a matrix that allows projecting the data in the initial space to one of the axis found by

ICA. The weight matrix is the full transformation from the original space. The independent component space may be expressed

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  International Journal of Innovative Research in Advanced Engineering (IJIRAE) ISSN: 2349-2163 

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as S = W X, where W is the weight matrix going from S space to the X space. X is the acquired data in the original space

and   S is the source activity. The rows of W are the vector with which we can compute the activity of one independent

component. To compute, the component activity in the formula S = W X, the weight matrix W is the mixing matrix. The activityof the Brain source S (also called “dipole”) is unit-less unless it is projected to the electrodes. Each dipole creates a voltage

signal at each electrode site, which can be measured. To re-project one component to the electrode space, we use X = W-1

S,where W

-1 is the inverse matrix to go from the source space S to the data space X. Rows of the S matrix are the time information

of the component activity and columns of the W-1 matrix are the scalp projection of the components. We denote the columns ofthe W

-1 matrix as the scalp topography of the components. Each column of this matrix is the topography of one component

which is scaled in time by the activity of the component. The scalp topography of each component can be used to estimate theequivalent dipole location for this component. These topo-plots are used to extract the most important five channels which have

shown maximum variation in the EEG amplitude.Topo-plot for one subject at the beginning and at the end of Pranayama for 20 seconds are plotted using EEGLAB Open

source software that runs on MATLAB Software and are shown in Figure 8a and Figure 8b respectively. 

Fig. 8a Topo-plot at the beginning of 1st stage of Pranayama

Fig. 8b: Topo-plot at the End of 3rd  stage of Pranayama

Blue to Red color variations in the Topo-plot indicate Electric Field potential variations from minimum to max value of EEG

signals represented as a vertical bar with + and – in the figure. By comparison of the plots during pre and post Pranayama stage,it is observed that there is a gradual increase in the field potentials in frontal and occipital regions towards the end of Pranayama

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for some channels. Channels 1, 8, 30, 31, 32 show more Electrical activity in frontal and occipital regions of the brain afterPranayama. These channels are used for extracting different EEG waves. By using elliptic filter, delta wave with a cutoff

frequency of 3.5 Hz, theta wave with a cutoff frequency of 7.5 Hz, alpha wave with a cutoff frequency of 11.5 Hz, beta with acutoff frequency of 30 Hz are extracted as shown in Figure 9. From the extracted EEG waves, by applying Fast FourierTransform (FFT) the power of all the waves is calculated. Finally, out of these five channels one channel which has got

maximum alpha power is considered for Analysis because Alpha wave is dominant when person is relaxed.

Fig. 9 Filtered EEG Data - Amplitude in µ Volts vs Number of Samples

III. EEG Amplifier Design and Data acquisition using MyDAQ: Biofeedback ApplicationTwo channel EEG Amplifier has been designed and tested for real time data acquisition of Biofeedback Application.

Biofeedback is a technique that measures Physiological functions and gives information about them, which in turn helps intraining and controlling the required Physiological functions. Biofeedback is most often based on measurements of Heart rate,Muscle tension, Skin conductivity and Brain waves. By watching these measurements, one can learn how to change these

functions by relaxing or by holding pleasant images or experiences in the mind. A sound or visual display may be used to knowwhen a goal or certain state is reached. Biofeedback teaches how to control and change these bodily functions. By doing so, onecan feel more relaxed or more able to cause specific muscle relaxation process. This may help treat undesirable conditions such

as Anxiety, Insomnia, Constipation, Tension and Migraine headaches. These observations help in the design of Bio feedbacksystem for self controlling alpha and beta level with external audio or video stimulus [14-18]. In this work, Biofeedback is

tested for brain waves using the Two-channel EEG Amplifier and MYDAQ System, design of which is presented in the nextsection.

IV. EEG Data Acquisition System Design Using MYDAQ and LABVIEW and ResultsIn the previous section, it was mentioned that FFT is used to compute the power of delta, theta, alpha and beta waves.

The square of the real part of FFT coefficient is the power. Before presenting the design of the EEG Data acquisition system, weneed to interpret the effect of Pranayama on subjects. The analysis of power variations would throw more light on the relaxationtrends of the subjects. Accordingly, the Percentage variations of alpha power in different regions of the brain among long term

 practitioners and the short term practitioners are acquired and presented in Figure 10 and 11 respectively. It may be observed thatthe Alpha power increases more for long term practitioners than for short term practitioners in Right occipital area of the brain,which is an indication that the corresponding subjects are in relaxed state. It may therefore be concluded that longer the practiseeffecting the Right occipital area, more relaxed the subject will be. It may also be noted that the left brain activity will be higherthan the right during logical thinking, reasoning etc.

Alpha and beta power may also be plotted for subjects in order to find percentage variation in relation to pre-meditationstate. Accordingly, power plots are presented in Figure 12 and 13 for various subjects during three Pranayama Stages presentedearlier in Figure 1. With respect to the Temporal context, at the end of the third stage of Pranayama, experiment conducted on

subjects showed that 40% of the subjects have increase in their alpha power, 50% have increase in alpha power in one stage and10% no change in alpha power. Further, 20% of the subjects showed no change in beta power, 30% showed decrease in beta power, 10% have increase in beta power in one cycle, and 40% showed increase in their beta power. These are shown in Figure

12 and 13 respectively. Therefore, we can conclude that 40 % of the subjects are relaxed as well as alert during the Pranayamastages.

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Fig. 10 Alpha Power Variation Percentage vs Long Term Practitioners

Fig. 11 Alpha Power Variation Percentage vs Short Term Practitioners

Fig.12 Alpha Power Variation of 10 Subjects During Three Pranayama Stages

Fig. 13 Beta Power Variation for 10 Subjects During Three Pranayama Stages

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Volume 1 Issue 9 (October 2014)  www.ijirae.com 

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Fig. 14 Two channel EEG Data Acquisition Setup Using MYDAQ

Fig. 15 Two channel EEG Data acquisition System Design Using MYDAQ and LABVIEW

Biofeedback Application

EEG Data acquisition setup is shown in Figure 14, where a couple of electrodes from the forehead of the Subject are

fed to the input of the EEG Amplifier, which in turn is connected to the MyDAQ of National Instruments. EEG data is acquiredfrom Pre-frontal positions FP3 and FP4. The acquired signals are displayed on the Laptop using LabView Design as presented inFigure 15. Acquired EEG Data is passed through second order Butterworth low pass Filter with appropriate cut-off frequenciesto extract alpha, beta, theta and delta waves from the Left and Right Pre-frontal positions. Power of Theta, Alpha and Beta forthese electrode positions are calculated by using FFT. The Threshold value is set at 50% of the baseline EEG of the subject inorder to light the indicators such as “Stabilised”, “2 fold increase” and “4 fold increase” optimally. This threshold need not be

50% and can be set as desired. For a particular cognitive task, if the power level of theta, alpha and beta value is constant, then itmeans no variations in the power level of EEG waves and therefore the “Stabilised” indicator lights up. If the power increasestwo times or 4 times then the respective indicator switches on. Subjects were asked to sit with eyes closed and instructed to

reduce the thoughts and to meditate for 5 minutes. Results show increase in alpha power in both left and right side of the brainafter meditation for 5 minutes as presented in Figure 16. With eyes open, and not performing any task, there was a decrease inalpha and beta power as shown in Figure 17. When the subject performed any Mathematical task in the mind such as counting

down a number task, then the beta wave power level increased as can be seen from Figure 18. We can see that the left side beta

 power increases when compared to that of the right, which is an indication that the Subject is in logical thinking state.

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Fig. 16 Theta, Alpha, Beta Levels after Performing Meditation

Fig. 17 Theta, Alpha, Beta Levels While the Subject is Resting With Eyes Open

Fig. 18 Theta, Alpha, Beta Levels While Performing Math Task with Eyes Open

IV. CONCLUSIONSAnalysis of EEG data during Pranayama practice shows increase of power in delta, theta, alpha and beta waves in long

term practitioners to a larger extent than short term practitioners. Spatial Analysis indicated higher variations in the EEG signals

in the right regions of the brain compared to the left regions of the brain. The Experimental results clearly show a higher level ofconsciousness experienced during Pranayama, which proves to be one of the best relaxation methods for cognitive enhancement,which helps the people to lead a stress free life. This paper presented the design of EEG Data Acquisition System using

MYDAQ and LABVIEW. This pilot study suggests a biofeedback protocol that might be useful in cultivating meditative statesof consciousness and providing real time feedback of enhanced alpha activity. There are now several mainstream health care programs which aid those, both sick and healthy, in promoting their inner well-being, especially mindfulness-based programs.More Research is required in this domain to prove to be of use for treatment of patients in Hospitals and rehabilitation centres.

Theta, Alpha and Beta power

Theta, Alpha and Beta power

Theta, Alpha and Beta power

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ACKNOWLEDGMENTSAuthors would like to thank all the volunteers for their support in conducting this study.

 

Experiment was conducted

with the consent of the Subjects.

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