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11-13 May, 2011, Aix-en-Provence, France easily. Under a low temperature (4℃) environment, we utilized an injection system to inject an NCD-4 solution with a molar concentration of 854.99 nM into the microenvironment. This allowed the NCD-4 molecules to connect with the MUA molecules in the sensing field. Then, a confocal microscope was used to examine the fluorescence signal of NCD-4 to determine the coverage condition of the MUA molecular layer. The wavelength of excitation light for NCD4 quantum dots is 405 nm and the wavelength of emitted light ranges from 400– 420 nm. (a) V. EXPERIMENT RESULTS This study used a confocal microscope to examine the adhesion condition of TYMV on sensing surfaces. The results were further analyzed with the image analysis software ImageJ. The ImageJ software was developed by the National Institutes of Health (NIH). The study also adopted ImageJ to analyze the surface area of the sensing surface. To prevent the MUA molecular layer coverage rate affecting the final TYMV adhesion effect, the MUA self-fabricated molecular layer was examined using NCD-4 fluorescent molecules as the samples. Utilizing observation by a confocal microscope and analysis of the results, Image J software was used to calculate the fluorescent coverage rate of the sensing field. In this manner, we located the MUA molecule layer with optimal coverage rate and uniformity. After the coverage rate and uniformity of the MUA molecule layer was confirmed, a TYMV adhesion experiment was conducted. Again, a confocal microscope was used to observe and analyze the adhesive condition. To ensure the accuracy of the experimental data, establishing a database for the control group was necessary. Fluorescent strength was defined as 100 A.U. Background fluorescent signals under 100 A.U. were ignored. The Au/MUA fluorescence testing revealed an absorption peak when the wavelength equaled to 415 nm. However, the peak value was 4A; hence, it did not affect the experiment when the wavelength was equal to 415 nm and can, therefore, be ignored. The degree of adhesion effect generated between Au and NCD-4 was then tested. When the wavelength equaled 410 mm, an absorption value of 260 A.U. was observed. This indicated that the autofluorescence of the two substances could influence the experiment result. Hence, future experiment statistics should filter the bright dots that have fluorescence strength of under 260 A.U. to avoid errors. After the control group data was established, we formed the MUA molecular layer by the dipping method, and then observed and analyzed by NCD-4. Fig. 7(a) shows that the MUA self-assembled monolayer (SAM) matured by the dipping method attributed to an MUA molecular layer that was unevenly formed and had low coverage. This is mainly because the molecules rely only on diffusion movement of Brownian motion as well as the effect of molecular aggregation. The result showed that fluorescence in the sensing field only accounts for 33.37 % of the total sensing area. Figure 7 (b) indicates that the average coverage rate of the MUA SAMs is approximately 86 % in the microenvironment when the rate of flow equals 1500 ml/hr. Fig. 7(a) MUA by dipping method (best viewed in color) (b) MUA when infusion rate of pump is 1500 ml/hr (best viewed in color) To overcome the problem of low coverage when implementing TYMV adhesion with the dipping method, the study adopted a microenvironment to improve the weakness of the dipping method. The autofluorescence of Au/MUA/EDC/NHS/MES is then examined and found that it cannot be ignored. Fluorescence strength under 180 A.U. should be filtered out to avoid errors in future experimental statistics. The results of confocal microscopy measurement are shown in the dipping method. In these results, the concentration of TYMV is 10ng/ml, and the adhesion duration is ten hours. Only a few TYMV with quantum dots are apparent on the sensor surface. The coverage rate of TYMV is poor with the dipping method. The average fluorescence strength is approximately 342.46 A.U. whereas the average fluorescence coverage is approximately 4.23 %. Additionally, some large molecules or particles are stuck on the surface which might have resulted from the aggregation of MUA, EDC, or NHS by the dipping method. This result is unfavorable for researchers that wish to detect viruses at extremely low concentrations in the solution. Figure 8 indicates the results seen after utilization of a microenvironment to enable TYMV to adhere to the sensing surface. We can see here that TYMV with modified NCD-4 has a concentration of 10ng/ml, and pump infusion rate was 1500ml/hr. The adhesion duration of TYMV was 1.4 minutes. The coverage was 70.1 % after analysis. When the fluorescence strength of X=2.5mm in Fig. 8 (the area directly behind the cylindrical structure) was analyzed, the area directly behind the cylindrical structure had a lower fluorescence strength while the sides of the structure revealed much higher fluorescence strength. The distribution is coincidentally identical with the curl size distribution that was simulated in this study. This phenomenon shows that the generation of curl contributes to the enhancement of the probability of viruses adhering to the sensing surface. The fluorescence strength is directly proportional to the size of curl in the position, as shown in Fig. 9. Figure 10 reveals that a microenvironment with vortex could certainly enhance the adhesion effect of TYMV to the sensing surface. The fluorescence coverage rate and unit duration of adhesion efficiency in the microenvironment with chaotic flow is respectively 16.5 times and 7,102 times greater than adhesion efficiency with the dipping method. The adhesion duration of TYMV in the microenvironment with chaotic flow is 0.0023 times less than that of dipping method. (b) 370
VI. CONCLUSION This paper discusses how to increase adhesive density of linkers and viruses on a sensor surface in microfluidic channels. We designed a flow movement in a microenvironment to control the adhesive density of MUA and TYMV. Adhesive density of a linker (MUA) and viruses (TYMV) with specific fluorescent dyes were measured by a confocal microscope. Our results show that TYMV and MUA layers disperse randomly by the dipping method. Infusion rate, flow rate, and vortex flow affect the adhesive density of the recognition layer on a sensor surface. An adhesion density of MUA was 86 % when the infusion rate was 1500ml/hr in the microenvironment. This was 2.57 times larger than the density detected by the dipping method. The virus, TYMV, could attain 70 % of adhesion densities when the infusion rate was 1500ml/hr in the microenvironment. The adhesion density was 16.5 times larger than the density detected by the dipping method. The duration of the experiment by vortex flow was 2.3×10 –4 times less than the duration by the dipping method. An interesting phenomenon was observed in that the fluorescence intensity distribution was similar to the vorticity distribution of simulation. Experimental results show that vortex flow method is able to increases the adhesive density of antigen-antibody reaction and it contributes to rapid and real-time detection. VII. ACKNOWLEDGMENT This paper is supported by the National Science Council, Taiwan. X Intensity (A.u.) in Y 11-13 May, 2011, Aix-en-Provence, France 1200 out Fig. 8 TYMV by vortex flow (best viewed in color) 4000 3500 3000 2500 2000 1500 1000 500 0 0 500 1000 1500 2000 2500 4000 3500 3000 2500 2000 1500 1000 500 0 Y position (m) Fig. 9 Vorticity and fluorescence intensity by vortex flow Vorticity (1/s) Average fluorescent intensity (A.U.) 1000 800 600 400 200 0 600 min Dipping method 1.7 min Vortex flow 100 Fig. 10 Average fluorescent intensity and coverage by dipping method and vortex flow REFERENCES [1] E. Engvall, P. Perlman, Enzyme-linked immunosorbent assay (ELISA). quantitative assay of immunoglobulin G, Immunochemistry, 8 (1971) 871–874 [2] R. M. Lequin, Enzyme immunoassay (EIA)/enzyme-linked immunosorbent assay (ELISA), Clin. Chem., 51 (2005) 2415–2418 (2005) [3] W. N. Burnette, Western blotting: electrophoretic transfer of proteins from sodium dodecyl sulfate - polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A, Anal. Biochem., 112 (1981) 195–203 (1981) [4] H. Towbin, T. Staehelin, J. Gordon, Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications, P. Natl. Acad. Sci. U.S.A., 76 (1979) 4350–4354 [5] D. B. Holt, P. R. Gauger, A. W. Kusterbeck, F. S. Ligler, Fabrication of a capillary immunosensor in polymethyl methacrylate, Biosens. and Bioelectron., 17 (2002) 95–103 [6] I. S. Park, N. Kim, Thiolated Salmonella antibody immobilization onto the gold surface of piezoelectric quartz crystal, Biosens. and Bioelectron., 13 (1998) 1091–1097 [7] C. W. Huang, G. B. Lee, A microfluidic system for automatic cell culture, J. Micromech. Microeng., 17 (2007) 1266–1274 [8] B. Steinhaus, M. L. Garcia, A. Q. Shen, L. T. Angenent, A portable anaerobic microbioreactor reveals optimum growth conditions for the methanogen, Appl. Environ. Microb., 73 (2007) 1653–1658 [9] Y. Gao, F. Y.H. Lin, G. Hu, P. M. Sherman, D. Li, Development of a novel electrokinetically driven microfluidic immunoassay for the detection of Helicobacter pylori, Anal. Chim. Acta., 543 (2005) 109–116 [10] P. Tabeling, Introduction to Microfluidics, Oxford University Press, New York, 2005, pp95-97 [11] H. Bruus, Theoretical Microfluidics, Oxford University Press, New York, 2008, pp79-81 [12] K. L. Bransom, J.J. Weiland, C.H. Tasi, T.W. Dreher, Coding density of the Turnip Yellow Mosaic Virus genome: roles of the overlapping coat protein and p206-readthrough coding regions, Virology 206 (1995) 403–412 [13] I. N. Serdyuk, N. R. Zaccai, J. Zaccai, Methods in Molecular Biophysics Cambridge University Press, New York, 2007, pp318-335 80 60 40 20 0 Average fluorescent coverage (%) 371
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COLLECTION OF PAPERS PRESENTED AT T
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III
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V
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Table of Contents Wednesday 11 May
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PANEL DISCUSSION TEXTILE MICROSYSTE
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Friday 13 May SESSION C4: APPLICATI
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SPECIAL SESSION OF BIO-MEMS/NEMS a
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suspended membrane can be pulled to
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most likely due to not yet consider
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that detectors may measure the diff
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2) Cavity width effect To verify th
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Fig.9. Capacitive sensor output, Va
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The anode and cathode are connected
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! TABLE 3 SUMMARY OF SELECTIVITIES
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The fabrication of optical Cap-Wafe
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the glass is polished down in a cos
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Fig. 14. Time history of the envelo
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We assume that 11-13 May 2011, Aix
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Y X Fig. 1. Scanning electron mic
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10 3 11-13 May 2011, Aix-en-Proven
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Intenstiy (counts/second) Fig. 1. X
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etween x 2 to L (remember that x 2
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piezoelectric displacement transduc
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Figure 7 Displacement conditions of
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protect the corners from undercut.
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A. Continuous domain Starting from
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Fig. 8. Central finite difference m
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700 11-13 May 2011, Aix-en-Provenc
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o o o o o o o o Check applicability
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C. Identification Results The ident
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The proposed solution for this prob
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teen wire segments of a coil, as in
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As the metallization is too reflect
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B. Implemented die overview A die c
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IN CLK OUT Fig. 16. Probe setup for
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adhesive material with 30μm thickn
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B. Dynamics of Energy Flows For a l
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80˚C. Additionally, we looked into
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The XRD shows a peak above the 2θ
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fibers which define the electric co
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Figure 15. Key board input system.
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ρ = h 2∆α∆T + + E 1h 3 1 +E 2
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In recent years, the pipe flow moni
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As the speed of the rotor increases
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inches silicon wafers which have th
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samples tested in different vacuum
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l c 11-13 May 2011, Aix-en-Provence
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Vp (V) 3,5 3,0 2,5 2,0 1,5 1,0 0,5
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II. MATHEMATICAL MODEL AND NUMERICA
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fluids travel along a curved channe
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measured mixing indices are depicte
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length, which enables them to focus
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however, a rotation for coincidence
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excludes the nature of the fixed bo
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Now the challenge in data handling
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value of every active channel. Ther
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electrocardiogram. • The heart be
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In our work, we proposed an innovat
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Fig. 8 Concept of the in-home perso
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found that the early-stage diagnosi
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Power monitoring data detected by w
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device consisted of a bimorph canti
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operation, the pressure inside the
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coefficient compatible with the mic
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Variable Description Value Crab-leg
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al. used a modified LIGA (German ac
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REFERENCES [1] G. Mehta, J. Lee, W.
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followed by titanium (Ti) which sho
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exposure dose, post exposure bake t
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#### ##/&### 3 # #
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*5%6,+/.,-,7-, ,+.,1/21* %
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B. Energy Applications Society is f
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Presently, the microfabrication pro
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[6] C. Richard, A. Renaudin, V. Aim
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NA sinθ c 11-13 May 2011, Aix-en-
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the waveguide on the fused silica,
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microphone diaphragm. The chosen CM
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TABLE V MICROPHONE CHARACTERISTICS
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Figure 7b shows the effect of a par
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