1. University of California, Irvine The Integrated Micro/Nano Summer Undergraduate Research Experience (IM-SURE) Single-Cell Platforms for Microbiomechanics Minh Guong Nguyen Biomedical Engineering University of California, Irvine   Mentor: Professor William C. Tang  Grad Student: Yu-Hsiang (Shawn) Hsu

2. OUTLINE Background –cytoskeleton –purposes Introduction –QCM –our piezoelectric transducer My responsibilities –design and develop experiments –collect and analyze resulting data Problems and future work

3. CYTOSKELETON COMPONENTS Intermediate filaments Fig. 1: Three types of protein filaments form the cytoskeleton Intermediate filaments MicrotubulesActin filaments protect cells and tissues from disintegration by mechanical stress essential component of cell division responsible for cell migration Alberts, Bruce, et al. Essential Cell Biology. 2nd ed. New York & London: Garland Science, 2004

4. ACTIN FILAMENTS Alberts, Bruce, et al. Essential Cell Biology. 2nd ed. New York & London: Garland Science, 2004 Fig. 2: Forces generated move a cell forward

5. WHY SINGLE-CELL PLATFORMS ? PURPOSE –mechanical changes of the cytoskeleton –parallel drug screening –cancerous cells identification and qualification –others

6. COMPARISON Traditional MethodOur method Fig 3: Sketch of the quartz crystal microbalance (QCM) experimental setup Fig. 4: A Single Cell Platforms for Microbiomechanics chamber cell Cannot detect 1 single cell mechanical property Not a precise result Detect 1 single cell mechanical property A precise result Jing Li, Christiane Thielemann, Ute Reuning, and Diethelm Johannsmann. “Monitoring of integrin-mediated adhesion of human ovarian cancer cells to model protein surfaces by quartz crystal resonators: evaluation in the impedance analysis mode.” BioSensors & BioElectronics 20 (2005): 1333-1340. Online posting. http://www.wctgroup.eng.uci.edu/http://www.wctgroup.eng.uci.edu/

7. EXPERIMENTAL SETUP Picture is taken by Minh Guong Nguyen, BME student, UCI The probe The Agilent 4395A

8. SiO2 ZnO CROSS SECTION OF OUR DEVICE Cross section of our device, drawing by Yu-Hsiang (Shawn) Hsu, Ph.D candidate, Dept of BME, UCI Si Au 200 µm Au cell

9. TOP VIEW OF OUR DEVICE Units in mm 200 µm in Diameter (our device) 1 mm square top electrode 15 µm thin lines Top view of our device, drawing by Yu-Hsiang (Shawn) Hsu, Ph.D candidate, Dept of BME, UCI

10. OUR DEVICE’S IMPEDANCES Resonance frequency Anti-resonance frequency Impedance (Ω) Frequency (MHz) Fig. 6: The graphs Impedance vs. Frequency of our device Data is collected by our experiments Impedance vs. Frequency Resonance frequency Anti-resonance frequency

11. THE QUALITY FACTOR Q M : The quality factor f a : The anti-resonance frequency (MHz) f r : The resonance frequency (MHz) Z r : The impedance at resonance frequency (Ω) C: The static capacitance (pF) http://www.morganelectroceramics.com/tutorials/piezoguide15.html

12. TABLE OF VALUES OF OUR DEVICES DeviceAnti-resonance frequency f a (MHz) Resonance frequency f r (MHz) Impedance at resonance frequency Z r ( Ω ) Static capacitance C (pF) 6-A5.5625.0811689.325 7-A5.4995.0851063.925 8-A5.5315.0941084.725 8-B5.5405.1121059.625 9-B5.5225.1031062.625 10-B5.5225.1031057.225 2-C5.5585.1031232.025 4-C5.5315.0941325.125 5-C5.5265.0851271.425 Data is collected by our experiments

13. THE QUALITY FACTORS (Q M ) OF OUR DEVICES The average is 7.3585 Calculation is based on our data obtained from experiments DeviceQuality factors (Q M ) 7-A8.1209 8-A 7.5990 8-B7.9203 9-B8.0410 10-B8.0911 2-C6.4500 4-C6.2203 5-C6.4259

14. COMPARISION OF OUR DEVICE WHEN TREATED WITH AND WITHOUT WATER The graph is based on our data collected form experiments Fig. 8: Comparison of our device when treated with and without water Frequency (MHz) Impedance (Ω) Impedance vs. Frequency Without Water With Water

15. DATA ANALYSIS DeviceResonance frequency f r (MHz) Anti- resonance frequency f a (MHz) Impedance at resonance frequency Z r ( Ω ) Viscosity (cP) Quality factor Q M Frequency shift Without water 3.61443.8141350.170.0185 (air) 4.9330.26 With water 3.60503.8366333.74 0.982 4.519 The frequency shift is related to the weight of water. The quality factor is related to the viscosity of water.

16. PROBLEMS AND FUTURE WORK Fig. 10: Graph of impedance vs. frequency Impedance (Ω) Impedance vs. frequency Frequency (MHz) Impedance vs. Frequency Impedance (Ω) Fig. 11: Graph of impedance vs. frequency B A D G O O DG O O D Noise interferes the signal

17. ACKNOWLEDGEMENTS Dr. William C. Tang Yu-Hsiang Hsu and John Lin Wyman Wong ALL FOR YOUR SUPPORT Said M. Shokair Edward M. Olano Sarah R. Martin UROP Fellows National Science Foundation

18. QUESTIONS?

19. Back up slide Comparison of our device when treated with and without water

20. Back-up slide Impedance (Ω) Frequency (MHz) The graphs of impedance vs. frequency of our devices zoom-in Impedance vs. Frequency ω = 2 () (f) Impedance of Capacitor: Z c = Impedance of Inductor: Z L = j ω L Impedance of Resistor: Z R = R

21. Butterworth-Van-Dyke (BVD) equivalent circuit Fig 6: The BVD equivalent circuit Inductor Resistor Capacitor Joachim Wegener, Jochen Seebach, Andreas Janshoff, and Hans-Joachim Galla. « Analysis of the Composite Response of Shear Wave Resonators to the Attachment of Mammalian Cells.» Biophysical Journal. Volume 78. June 2000: 2821-2833. Fig. 7: Lumped-element equivalent circuit