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1323 International Journal of Advanced Research in Computer Engineering & Technology Volume 1, Issue 5, July 2012 Micro-cantilever Sensors for Detection of Pesticide Contents in the Water-table of Malwa Region in Punjab Gurleen Kaur, Ravinder Singh Sawhney, Rajan Vohra Department of Electronics Technology, Guru Nanak Dev University, Amritsar Email: gurleen_gnps2000@yahoo.com; Sawhney_gndu@hotmail.com; sargam_111@yahoo.co.in Abstract- Biosensors based on microcantilevers have become a Micromachined cantilevers were first used as force probes in promising tool for directly detecting biomolecular interactions Atomic Force Microscopy (AFM). Their extreme sensitivity with great accuracy. Microcantilevers translate molecular to several environmental factors, such as noise, temperature, recognition of biomolecules into nanomechanical motion that is humidity and pressure was immediately evident. In 1994, commonly coupled to an optical or piezoresistive read-out detector system. Biosensors based on cantilevers are a good research teams in Oak Ridge National Laboratory and IBM, example of how nanotechnology and biotechnology can go converted the mechanism causing interference into a platform together. High-throughput platforms using arrays of cantilevers for a novel family of biosensors. By measuring the change in have been developed for simultaneous measurement and read- resonance frequency, microcantilevers were shown to be out of hundreds of samples. As a result, many interesting sensitive to mass changes, with a better yield than applications have been develpoed and the first sensor platforms piezoelectric gravimetric conventio-nal sensors. The are getting commercialized. In this paper, we propose the laboratories with MFA instruments displayed a substantial application of Microcantilever sensor for the detection of interest in cantilevers as a new platform for a variety of pesticide contents in the water table of the Malwa region in chemical and physical biosensors. Punjab (infamous as the cancer belt of the region) due to environmental toxicity following the excessive use of pesticides and fertilizers under the disguise of Green Revolution. The results concluded here can be applied in general to the various parts of India, where use of the pesticides is excessive and that is causing the adulteration of the water table. Keyword— Microcantilever, Biosensor, NEMS, Biomolecular interaction I. INTRODUCTION Microcantilevers are micromechanical beams that are anchored at one end, such as diving spring boards those can be readily fabricated on silicon wafers and other materials. Their typical dimensions are approximately 100 microns long, 20 microns wide and 1 micron thick. The microcantilever sensors are physical sensors that respond to surface stress Figure1. (a) Schematic diagram of a microcantilever indicating dimensions changes due to chemical or biological processes [1]. When length, L, resistor length, LR, width, w, and thickness, h. (b) An array of fabricated with very small force constants, these can measure piezoresistive cantilevers. (c) Cross-sectional diagram through the layers of the microcantilever sensors [3] forces and stresses with extremely high sensitivity. The very small force constant (less than 0.1 N/m) of a cantilever allows With recent advancements in technology, microcantilever detection of surface stress variation due to the adsorption (or sensors have found wider applications in the field of specific surface-receptor interaction) of molecules [2]. medicine, specifically for the screening of diseases, detection Adsorption of molecules on one of the surfaces of the of point mutations, blood glucose monitoring and detection of typically bimaterial cantilevers (silicon or silicon nitride chemical and biological warfare agents. These sensors have cantilevers with a thin gold layer on one side) results in a several advantages over the conventional analytical differential surface stress due to adsorption-induced forces, techniques in terms of high sensitivity, low cost, simple which manifests as a deflection [3]. In addition to cantilever procedure, low analyte requirement (in µl), non-hazardous bending, the resonance frequency of the cantilever can vary procedures and quick response. Moreover, the technology has due to mass loading. These two signals, adsorption-induced been developed in the last few years for the fabrication and cantilever bending when adsorption is confined to one side of use of nanocantilevers for sensing applications, thereby giving the cantilever and adsorption-induced frequency change due rise to nanoelectromechanical systems (NEMS) [8]. This to mass loading, can be monitored simultaneously [2-7] development has increased the sensitivity limit up to the Capacitance, piezoresistance and resonance frequency are extent that researchers can now visualize the counting of among the sensing principles depending upon the mechanical molecules. properties of the device. 285 All Rights Reserved © 2012 IJARCET
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1323 International Journal of Advanced Research in Computer Engineering & Technology Volume 1, Issue 5, July 2012 Microcantilever sensors are most promising for microbiosensors and nanobiosensors. This new class of highly sensitivity biosensors can perform local, high resolution and label-free molecular recognition. The microcantilevers translate the molecular recognition of biomolecules into nanomechanical motion [9] (from a few nm to hundreds of nm), which is commonly coupled to an optical or piezoresistive read-out system [10,11]. Research in this new type of sensor is growing exponentially after the landmark paper of Fritz et al. in 2000, in which the ability of microcantilever sensors is shown in discerning single-base variations in DNA strands without using fluorescent labels. Shortly afterwards, microcantilever sensors were also shown to work in DNA hybridization [11] and detection of proteins involved in cancer [12] and other diseases [13] with increased accuracy, as well as in environmental sciences [14].Cantilever This type of surface tension induced by molecular interactions sensors have also been used for detecting chemicals, such as is not generally observed on the surface of common materials. volatile compounds [15], warfare pathogens [16], explosives The cantilever senses the tension and bends in response to the [17], and glucose [18], and ionic species, such as calcium ions free energy changes taking place at its surface. [19]. II. THEORY OF OPERATION A microcantilever is a device that can act as a physical, chemical or biological sensor by detecting changes in cantilever bending or vibrational frequency. It is the miniaturized counterpart of a diving board that moves up and down at a regular interval. This movement changes when a specific mass of analyte is specifically adsorbed on its surface similar to the change when a person steps onto the diving board. But microcantilevers are a million times smaller than the diving board having dimensions in microns and different shapes as shown in figure 2. Molecules adsorbed on a microcantilever cause vibrational frequency changes and deflection of the microcantilever. Viscosity, density, and flow rate can be measured by detecting changes in the vibrational frequency. Another way of detecting molecular adsorption is by measuring deflection of the cantilever due to adsorption stress Figure 2: Different types of microcantilevers (top view) on just one side of the cantilever. Depending on the nature of (a) Rectangular (b) Double-legged (c) Triangular [19] chemical bonding of the molecule, the deflection can be up or down. Biochips with mechanical detection systems commonly The immobilization of molecules on the cantilever surface is use microcantilever bi-material (e.g. Au–Si) beams as sensing required for its use as a nanomechanical sensor (Figures 3 and elements. The Au side is usually coated with a certain 4). The selection of the molecule depends on de intended receptor. Upon the binding of the analyte (e.g. biological application. This principle applies whether if the molecule to molecules, such as proteins or biological agents) with the be detected (analyte) will be presented in a solid, liquid or a receptor, the receptor surface is either tensioned or relieved. gas phase. The immobilized molecules provide the cantilever This causes the microcantilever to deflect, usually in with specificity for the analyte. The specific molecular nanometers, which can be measured using optical techniques. interactions taking place at the flexible surface of cantilever The deflection is proportional to the analyte concentration. increase surface tension forcing the cantilever to bend [1]. The concept has been employed in screening certain diseases All Rights Reserved © 2012 IJARCET 286
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1323 International Journal of Advanced Research in Computer Engineering & Technology Volume 1, Issue 5, July 2012 such as cancer and detecting specific chemical and biological case of tensile stress) the surface area as shown in figure 4. If warfare agents. this stress is not compensated at the opposite side of a thin plate or beam, the whole structure will bend. Between the III. MECHANICAL PROPERTIES OF CANTILEVERS areas of compressive stress and tensile stress, there is a neutral plane which is not deformed. Due to bending, a force F is The basic mechanical parameters of a cantilever are the spring acting at a distance of x in the neutral plane results in a constant and the resonance frequency. The spring constant k bending moment M=F.x. Therefore, the radius of curvature R is the proportionality factor between applied force, F and the is given by: resulting bending of the cantilever, z. This relation is called Hooke’s law. 1/R = d2z/dx2 = M/EI F = -kz where E is the apparent Young’s modulus and I is the moment of inertia given by the following equation for rectangular The spring constant yields the stiffness of the cantilever. For a beams rectangular cantilever of length l, the spring constant can be written as The change in the surface stress at one side of the beam will cause static bending, and the bending moment can be where E is the Young’s modulus and I is the moment of calculated as: inertia. A typical spring constant for a stress sensitive cantilever is in the range of 1 mN/m to 1 N/m. The resonance frequency fres for a simple rectangular cantilever can be expressed as Δσ = σ1 – σ2 is the differential surface stress with σ1 and σ2 as surface stress at the upper and lower side of the cantilever respectively (figure 5). Inserting these values of I and M in the first equation yields Stoney’s formula where ρ is the mass density, h and w denotes the height and the width of the cantilever respectively. The moment of inertia for a rectangular cantilever can be written as A simpler expression for the resonance frequency can be written as a function of the spring constant as Fig.5 Bending of a cantilever beam in response to compressive and tensile stresses. (a) Compressive surface stress due to repulsion between the where mass, m=ρ. h. l. w. The relation shows that the biomolecules leads to downward/negative deflection of the cantilever beam. resonance frequency increases as a function of increasing (b) Tensile surface stress due to attraction between molecules leads to spring constant and of decreasing cantilever mass. [20] upward/positive deflection of the cantilever beam. IV. MODALITIES OF CANTILEVER DEFLECTION- Changes in surface stress can be the result of adsorption BASED SENSING process or electrostatic interactions between charged molecules on the surface as well as changes in the surface A uniform surface stress acting on an isotropic material hydrophobicity and conformational changes of the adsorbed increases (in the case of compressive stress) or decreases (in molecules. In addition to surface stress-induced bending, the volume expansion of bimaterial cantilevers can result in a All Rights Reserved © 2012 IJARCET 287
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1323 International Journal of Advanced Research in Computer Engineering & Technology Volume 1, Issue 5, July 2012 static bending. A bimaterial cantilever undergoes bending due Substances at trace levels are currently detected by various to gas adsorption if the volume expansion coefficients of the techniques like high performance liquid chromatography two materials are different. (HPLC), thin layer chromatography (TLC), gas chromatography (GC), gas liquid chromatography (GLC) etc. V. APPLICATIONS However, these techniques are complex, time consuming, costly and require bulky instrumentation. The surface of the cantilever should be coated with a layer of detector molecules that can react with the analyte and detect Also sample preparation is a prolonged complex procedure biochemical reactions at its surface. This novel detection and requires skilled personnel. But the microcantilever-based technique based on cantilevers is extremely sensitive. sensors can detect trace amounts of substances in parts-per Cantilever based biosensors have enormous potential, billion (ppb) and parts-per-trillion (ppt). They translate especially in the field of biochemical analyses. Fast and biomolecular recognition into nanomechanical bending of the simple biochemical detectors based on this detection method microcantilever [23]. Intermolecular forces arising from the can be constructed. Studies of simple molecular interactions adsorption of analyte molecules onto the microcantilever would also be feasible due to the exquisite mechanical induce surface stress, directly resulting in nanomechanical sensitivity of micro - cantilever (Figure 6). The technology of bending of the microcantilever. cantilever biosensors is applicable to a number of specific VII. PROPOSAL tasks [21]: Punjab state Health Minister Laxmi Kanta Chawla had admitted during Assembly Session on March 7, 2010 that between 2001 and 2009, the total number of cancer patients detected in the rural areas numbered 23,427, of which 16,730 died. Excessive use of pesticides, chemical fertilizers and uranium are being incriminated for making the environment and food chain highly toxic. The surface water as well as ground water has been grossly contaminated by the above Figure 6: Application of cantilever to the detection of superficial changes (a) mentioned sources of toxicity. The Malwa area of Punjab has tension (b) temperature and (c) mass [21] emerged as the epicenter of the disease and has come to be 1. Life Sciences: For studying the bases of the interaction known as the cancer belt of Punjab. among biological molecules. To develop novel analyses based on this platform with a potential applicability to portable A study undertaken by an NGO has indicated that drinking devices. water being supplied particularly in the Malwa belt is source 2. In vitro diagnostics: To develop faster, more sensitive and of a cocktail of toxins as it was a combination of pesticides, label free methods (no tracers are needed for final nitrates, heavy metals and fluoride. Possibly this cocktail was identification) to analyze chemical or biological samples. causing the whole spectrum of ill health in Punjab including 3. Drug discovery: To study the interactions between small Cancer [24]. Uranium has also been found in the water molecules and their specific receptors. To conduct samples beyond the permissible limits in groundwater in the multiplexed analyses increasing the parallelism and contents Malwa region. Uranium alone or along with other components of the results. of the toxic cocktail may be causing mental retardation, 4. Fresh water control: To detect heavy metal ions in fresh physical deformities and neurological problems among water. To develop assays that could be useful to implement a children in Punjab and surrounding regions [20]. decentralized system to monitor the quality of the resins, or Keeping the above scenario in mind, there is an urgent need other chemicals used in the fresh water distribution system for the sensitive detection of the abnormalities in the soil [22]. content. The use of specially fabricated microcantilevers can serve our purpose here. Ultrasensitive microcantilevers can act as precise sensors if we measure their nanomechanical responses in different environments. Microcantilevers can be VI. ADVANTAGES used to detect the pH of the soil and hence obtain its salinity. It has been spotted that when an intelligent hydrogel is Microcantilever based sensors have enormous potential for the applied to the surface, to detect the deflection of a detection of various analytes in gaseous, vacuum and liquid microcantilever, it swells at variable pH levels. The hydrogel medium. They have aroused considerable interest because of was patterned onto microcantilevers of various sizes using their high specificity, high sensitivity, simplicity, low cost,low UV photolithography. When the cantilevers were soaked in analyte requirement (in µl), non-hazardous procedure with different buffers, the swelling hydrogel induced surface stress fewer steps, quick response and low power requirement. All Rights Reserved © 2012 IJARCET 288
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1323 International Journal of Advanced Research in Computer Engineering & Technology Volume 1, Issue 5, July 2012 and therefore the bending phenomenon. The deflection was Microcantilevers have got potential applications in every evident, and differed accordingly at various pH levels [25]. field of science ranging from physical and chemical sensing to biological disease diagnosis. The major advantages of Heavy-metal ions and ions in general have also been studied. employing microcantilevers as sensing mechanisms over the Ji et al. used thiol-derivatized calixarene and crown-ether conventional sensors include their high sensitivity, low cost, macrocycle-functionalized cantilevers to detect Cs2+ ions in low analyte requirement (in µl), non-hazardous procedure the range 1011–107 M and K+ in the 104 M range [26]. with fewer steps (obviating the need for labels), quick Following the same grounds amount of Uranium can be response and low power requirement. detected in the ground water of Malwa region using The technology holds the key to the next generation of micromechanical sensors. highly sensitive sensors. To sum up, microcantilever-based biosensors comprise a continually growing novel technology Other functionalization schemes have shown that cantilevers and, because of their great capabilities, offer an alternative to were able to detect, with great accuracy and selectivity, current biosensor technologies. Cantilevers will play an different ions, such as CrO4-2 [27], Ca2+17 or Pb2+ [28]. So essential role in the immediate future of nanobiotechnology. excessive fertilizer contents like NO3- and PO4- can be checked in soils by application of cantilever sensor REFERENCES technology. Nanomechanical cantilever sensors can be the most sensitive devices for pesticide detection using [1] Nanomechanical Biosensors: a new sensing tool. L.G. Carrascosa, M. Moreno, M. A´ lvarez, L.M. Lechuga;Trends in Analytical Chemistry, immunoreactions due to the tiny reaction area (/100 mm2) Vol. 25, No. 3, 2006 . compared with another label-free biosensors such as the [2] Gimzewski, J. K.; Gerber, C. H.; Mayer, E.; Schlitter, R. R. Chem.Phys. surface plasmon resonance biosensor (mm2) and the quartz Lett. 1994, 217, 589. crystal microbalance (cm2) Cantilever based assays for [3] Chen, G. Y.; Thundat, T.; Wachter, E. A.; Warmack, R. J. J. Appl. Phys. 1995, 77, 3618. pesticide detection has been reported [29,30]. The stress [4] Gimzewski, J. K.; Gerber, C.; Meyer, E.; Schlittler, R. R. NATO ASI induced by the binding of a pesticide residue BAM (2,6 Ser., Ser. E: Appl. Sci. 1995, 286, 123. dichlorobenzamide) immobilized on a cantilever surface to [5] Thundat, T.; Chen, G. Y.; Warmack, R. J.; Allison, D. P.; Wachter,E. A. anti-BAM antibody is measured using with four gold-coated Anal. Chem. 1995, 67, 519. [6] Thundat, T.; Wachter, E. A.; Sharp, S. L.; Warmack, R. J. Appl.Phys. cantilevers and piezo resistive . Lett. 1995, 66, 1695. Development of nanomechanical biosensors for detection of [7] Wachter, E. A.; Thundat, T. ReV. Sci. Instrum. 1995, 66, 3662. the pesticide DDT has also been cited. Detection of the [8] R. McKendry, J. Zhang, Y. Arntz, T. Strunz, M. Hegner, H.P. organochlorine insecticide compound Lang,M.K. Baller, U. Certa, E. Meyer, H.-J. Guntherodt, C. Gerber, Proc.Natl. Acad. Sci. U.S.A. 99 (2002) 9783. dichlorodiphenyltrichloroethane(DDT) by measuring the [9] J. Fritz, M.K. Baller, H.P. Lang, H. Rothuizen, P.Vettiger, E. Meyer,H.- nanometer-scale bending of a microcantilever produced by J. Gu¨ ntherodt, C. Gerber, J.K. Gimzewski, Science (Washington)D.C. differential surface stress. A synthetic hapten of the pesticide 288 (2000) 316. conjugated with bovine serum albumin (BSA) was covalently [10] R. Raiteri, M. Grattarola, H.-J. Butt, P. Skla´dal, Sens. Actuators, B79 (2001) 115. immobilised on the gold-coated side of the cantilever by using [11] P.G. Datskos, T. Thundat, N.V. Lavrik, Encyclopedia thiol self assembled monolayers [31]. The immobilisation Nanosci.Nanotechnol. 10 (2004) 1. process is characterised by monitoring the cantilever [12] G. Wu, R.H. Datar, K.M. Hansen, T. Thundat, R.J. Cote, deflection in realtime. Then specific detection is achieved by A. Majumdar, Nat. Biotechnol. 19 (2001) 856. [13] C.A. Savran, S.M. Knudsen, A.D. Ellington, S.R. Manalis, Anal.Chem. exposing the cantilever to a solution of a specific monoclonal 76 (2004) 3194. antibody to the DDT hapten derivative. The specific binding [14] M. Alvarez, A. Calle, J. Tamayo, L.M. Lechuga, A. Abad,A. Montoya, of the antibodies on the cantilever sensitised side is measured Biosens. Bioelectron. 18 (2003) 649. with nanomolar sensitivity. Direct detection is proved by [15] M.K. Baller, H.P. Lang, J. Fritz, C. Gerber, J.K. Gimzewski,U. Drechsler, H. Rothuizen, M. Despont, P. Vettiger,F.M. Battiston, J.P. performing competitive assays, in which the cantilever is Ramseyer, P. Fornaro, E. Meyer,H.J. Guntherodt, Ultramicroscopy 82 exposed to a mixed solution of the monoclonal antibody and (2000) 1. DDT [32]. [16] B.L. Weeks, J. Camarero, A. Noy, A.E. Miller, L. Stanker,J.J. De Yoreo, Scanning 25 (2003) 297. SUMMARY [17] L.A. Pinnaduwage, A. Gehl, D.L. Hedden, G. Muralidharan,T. Thundat, R.T. Lareau, T. Sulchek, L. Manning, B. Rogers,M. Jones, J.D. Adams, In this review article, we have provided an overview of most Nature (London) 425 (2003) 474. technical aspects of the new microcantilever-based sensors [18] A. Subramanian, P.I. Oden, S.J. Kennel, K.B. Jacobson, which is a promising technology and emerging as a suitable R.J. Warmack, T. Thundat, M.J. Doktycz, Appl. Phys. Lett. 81(2002) solution for important problems. Improvements in 385. [19] Cantilever technology. 2006.[Sitio en Internet]. Disponible en: reproducibility and sensitivity, and integration of http://www. Cantion.com/cantilever_technology.htm. microfluidics and detection systems are the main aims of current research. 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1323 International Journal of Advanced Research in Computer Engineering & Technology Volume 1, Issue 5, July 2012 [20] Microcantilevers: Sensing Chemical Interactions via Mechanical In tern ation al and Nation al Co mp u ter Societies. His key areas Motion Karen M. Goeders, Jonathan S. Colton, and Lawrence A. of interest in the f ield of Data Co mmu n ication are routing Bottomley th rough WLANs, MANETS, Mesh Networks, MPLS, Ant [21] Nanomechanical cantilever. 2006.[Sitio en Internet]. Disponible en: Colon y Op timizatio n, Particle Swarm Optimization and http:// www.Cantion.com/applications.htm .Último acceso:12 octubre 2006 Diff erent Encryp tion Techn iqu es. He h as been on th e board of [22] A Review of Microcantilevers for Sensing Applications Sandeep Kumar man y jou rnals as rev iewer. Curren tly, h e is wo rking in th e Vashist, http://www.azonano.com/oars.asp field of mo d ell ing and si mu lation of ch arge tran spo rt th rough [23] Pesticide Pollution in Punjab: A Review N.S. Tiwana, Neelima v ario us mo lecu les and d esigning of v arious mo lecu lar Jerath*, Gurharminder Singh and Ravleen Singh Asian Journal of jun ctions fo r futu re Nano electron ic Dev ices . Water, Environment and Pollution, Vol. 6, No. 1, pp. 89-96. [24] Ramaninder K Bhatia, Cancer incidence on rise in farm animals too? Er. Rajan Voh ra has been wo rking as - The Times of India, Feb 10, 2011 Asstt. Professo r in th e Dep artmen t of [25] Amit Vasudev, amitv@stanford.edu, Microelectronics Research Center, Electro nics Techno log y, Gu ru Nan ak D ev The University of Texas at Austin, NNIN REU 2006 Research University, Amritsar . He has exp erien ce Accomplishments of two years of teach ing post -g radu ate as [26] H.-F. Ji, R. Dabestani, G.M. Brown, P.F. Britt, Chem. Commun.(Cambridge) (6) (2000) 457 well as und erg radu ate eng ineering [27] H.-F. Ji, T. Thundat, R. Dabestani, G.M. Brown, P.F. Britt,P.V. stud en ts. He has a k een in terest in data Bonnesen, Anal. Chem. 73 (2001) 1572. co mmu n ication, esp ecially v arious [28] K. Liu, H.-F. Ji, Anal. Sci. 20 (2004) 9. app lications based on Wireless Lo cal Area Netwo rk s and [29] Alvarez M, Calle A, Tamayo J, Lechuga LM, Abad A, Montoya A. pub lish ed mo re th an 15 p ap ers in v arious In tern ation al jou rnals Development of nanomechanical biosensors for detection of the and In ternation al & National co nfe ren ces. He h as been pesticide DDT. Biosens Bioelectron. 2003;18:649–653. doi: no min ated fo r th e f in als for th e award of young scientist 10.1016/S0956-5663(03)00035-6. du ring 1 5 t h Pun jab scien ce Cong ress. He has me mb ersh ip of [30] Suri CR, Kaur J, Gandhi S, Shekhawat GS. Label-free ultra-sensitive v ario us In tern ation al and national so cieties like detection of atrazine based on IACSIT,IAOE,CSTA etc. nanomechanics. Nanotechnology. 2008;19:235502. doi: 10.1088/0957-4484/19/23/235502. [31] Bruun L, Koch C, Pedersen B, Jakobsen MH, Aamand J. A quantitative enzyme-linked immunoassay for the detection of 2,6- dichlorobenzamide (BAM), a degradation product of the herbicide dichlobenil. J Immunol Methods. 2000;240:133–142. doi: 10.1016/S00221759(00)00190-3. [32] Development of nanomechanical biosensors for detection of the pesticide DDT, Mar Alvarez a, Ana Calle a, Javier Tamayo , Laura M. Lechuga a, Antonio Abad , Angel Montoya. Gurleen Kaur is a student of B.tech in Department of Electronics Technology, Guru Nanak Dev University, Amritsar. She is an all rounder and a bright student. She has an outstanding academic career since school times. She represented India in Asia Region Space Settlement Design Competition (ARSSDC), NASA as team leader in 2008 and obtained a gold in National Aerospace Olympiad held at Nagpur in 2009. Though she has been studying in B.Tech Vth Semester, She has already a few publications to her credit. Prof. Rav ind er Singh Sawhney has been wo rk ing as facu lty with th e Departmen t of Electro nics Tech nolo g y, Gu ru Nanak Dev Un iv ersity, Amritsar. He has teachin g exp erien ce of mo re than 15 years to bo th po st -g radu ate and un der g radu ate en gin eering stud ents. He has mo re th an 35 pub lication s to h is cred it in v ario us In ternation al jou rn als as well as In tern ation al and Nation al co nf eren ces. He h as me mb ersh ip of man y All Rights Reserved © 2012 IJARCET 290
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