CAMD 3D Bio-MEMS device to detect Salmonella Bacteria

Transcription

1 3D Bio-MEMS device to detect Salmonella Bacteria Flavio Aristone UFMS: Federal University of South Mato Grosso : Center of Advanced Microstructures and Devices LSU: Louisiana State University

2 Co-workers Jost Goettert, Yohannes Desta, Lin Wang, Jin Yoonyoung, Proyag Datta, Dawit Yemane, Tracy Morris, Shaloma Malveuax, Changeng Liu, Kun Lian, Zhong-Gen Lin, Joseph Kouba, Antoine Dupuy, Niko Baether, Alexey Espindola, Thomas Mueller, Jakob Weimnert 2

3 OUTLINE General Concepts Examples of Micro-systems Summary LiGA Lithography; X-ray masks Exposures Electroplating; Hot-embossing Detection of Salmonella Bacteria Biological Protocol Devices and Tests Conclusions 3

4 Miniaturization By miniaturizing applications you can... enhance performance and productivity reduce sample and reagent consumption work more easily with nanoliter sample volumes 4

5 Integration By integrating multiple steps into a single streamlined process you can... increase productivity improve reproducibility eliminate the need for user intervention minimize the risk of losing samples 5

6 Micro Harmonic Drive Gear Flexspline Output flange Output bearing Flexspline Planet gear Dynamic Spline Circular Spline Dynamic Spline Motor Dynamic Spline Circular Spline Sun gear Flexspline Housing Planet gear Planet gear 6

7 Advantages Zero backlash yet miniature dimensions Excellent repeatability High torque capacity High reduction ratios with 6 parts High efficiency Extremely flat design Very low weight 7

8 Applications for Micro-Motors Medical Equipment Semicon Laser Equipment Optical Communication Measuring Machines Biotechnology Robotics Aircraft Microscopes Spacecraft 8

9 Bio-MEMS Biotechnology Medical Equipment 9

10 Microfluidics 10

11 High Aspect Ratio µ-gc Gas Chromatograph - Drawing Board to Plastic Microstructures Nickel GC mold insert fabricated using LIGA Drawing of a micro gas chromatograph (GC) SEM picture of a small section of the nickel micro GC mold insert Microstructures molded in plastic using LIGA fabricated GC mold tool. The SEM pictures show various sections. These structures are 50 microns wide and 420 microns high. 11

12 Some examples thickness = 500 microns; column diameter = 100 microns; turn diameter = 75 microns thickness = 500 microns; column diameter = 100 microns; turn diameter = 100 microns 12

13 GC 13

14 Summary Microfluidic lab-on-a-chip technology represents a revolution in laboratory experimentation. It combines manufacturing methods from the microchip industry with expertise in fluid dynamics, biochemistry and software and hardware engineering to develop miniature, integrated biochemical processing platforms, systems, and instrumentation. The benefits of miniaturization, integration and automation will strengthen research-based industries and may also lead to new point-of-care medical and analytical devices. 14

15 To keep in mind! Let technology NOT drive you towards complete miniaturization at the start! Stay focused to fabricate a product! Develop strategic partnerships! Establish a multi-disciplinary TEAM effort! 15

16 The beginning Late 70 s/early 80 s Development of concept to fabricateseparation nozzles for uranium enrichment at IKVT/KfK (today IMT/FZK) U 235/238 Many thousands of nozzles cascaded in an array U 235 Tip ~ 3µm Height ~ 300 µm U

17 LiGA Process L i :Lithography G : Electroforming A : Molding 17

18 Microfabrication 1. Idea / Project 2. Drawing 3. Optical Masks 4. X-Ray Masks 5. Substrate preparation 6. Exposure 7. Development 8. Electroplating 9. Mold insert final work 10. Device replication 11. Testing 12. Results / Analysis 18

19 Lithography Shadow printing using x-rays X-ray mask Resist Substrate Development 19

20 Electroforming and Molding Electroplating of metal structures and mold inserts Replication by molding (hot embossing, injection molding) 20

21 X-ray masks Membrane Absorber (5-30 µm) Desirable mask membrane characteristics Good X-ray and optical transmission Good mechanical stability, low internal stress Radiation resistant Compatible with established mask making processes and equipment Compatible with plating of Au absorber Thin membranes: Silicon, Silicon Nitride, Titanium, Diamond Frame Thick low Z substrates: Graphite, Beryllium, Silicon, glass 21

22 Mask fabrication E-beam lithography UV lithography Electroplate µm of Au Photomask Soft X-ray lithography Intermediate X-ray mask Electroplate 5-50 µm of Au Electroplate 5-50 µm of Au Working X-ray mask Working X-ray mask 22

23 Thin membrane Silicon etching setup for making 1µm thick Silicon Nitride Membrane Condenser Water circulation Magnet stir Hotplate Window size mm 23

24 Properties of 2500 µm tall templates for micro-gears Sub-micrometer structural details NiFe gear assembly Extreme structure heights 24

25 LiGA Microstructures Wide variety of materials possible non-vertical sidewalls Arbitrary cross-sectional shape Smallest structures of some micrometer Vertical and smooth sidewall multi-level pattern 25

26 Patterning Accuracy - DXRL Deviation from perfect sidewall ~ 0.06µm / 100 µm Ph.D. Thesis Jürgen Mohr, IMT/FZK,

27 Mold insert 27

28 Overplating 28

29 Nickel Mold insert 29

30 Hot embossing A thermo-plastic replication process suitable for LiGA HARM structures. Key process steps include heating, evacuating, stamping, and demolding. 30

31 Replication process Insert plastic, close press, evacuate, and heat up. Apply high pressure to transfer structures. Cool down, vent, and demold structured plastic parts. 31

32 LiGA Process 32

33 Definition To make things clear. XRL or soft XRL: X-ray lithography in thin resists DXRL: Deep X-ray lithography (up to 1mm) UDXRL: Ultra-deep x-ray lithography (above 1mm) LiGA is not to make the next generation micro-chips. It is a ultra-precision micromachining process using lithographic tools! 33

34 LSU a Synchrotron Radiation Facility Dedicated to Microfabrication 34

35 Synchrotrons in the U.S. SRC ALS SSRL APS CHESS NSLS SURF 35

36 Synchrotron Radiation Storage Ring Shield Wall Synchrotron radiation is electromagnetic radiation (light) emitted from electrons (positrons) moving with relativistic velocities on macroscopic circular orbits. 36

37 Lightening the sample 4 Synchrotron lines operating at GeV for µfab 1 for SOFT X-rays from Cr double mirror system 1 for HARD X-rays from 7 T Wiggler Vacuum safety S h ie ld W a ll G V1 D IP O L E V A C TA N K BSS D ia g n o s t ic F la g G V2 Pu m p P o rt Pu m p P o rt BEAM PS ( ) G V3 FS F ilt e r C h a m b e r B e W in d o w Sca nner IP IP IP ~ 7 5 t o S. P. Heat protection ~ t o M a s k P la n e Radiation safety 37

38 Some Infrastructure at Oxford ion milling Inside the clean room Quintel UV aligner Temescal e-beam 26 de junio de 2004 evaporator Bariloche - Argentina Plating Station 38

39 and more Infrastructure Resist Press Metrology Material characterization Chemistry lab Surface finishing Molding SEM/EDAX Micro hardness testing SPM HEX 02 Hot Embossing Machine 26 Lapper de junio de 2004 Veeco RST NT3300 DSC, DMA, TGA, TMA Material testing 39

40 Concept of 3D µ Fluidic-S 40

41 Modular Test Structure Molded Modular Micro-fluidic Structure 41

42 Multilevel Microfluidic Device 42

43 Result 43

44 Lab-on-a-CD Centrifugal Bio-Chips For the Analysis of Biologically Fluid Samples Centrifugal Bio-Chips For the Analysis of Biologically Fluid Samples G-vector Rotates 180o in vertical plane Spindle Rotates 360o in horizontal plane Collaboration: Mark F. Clarke, University of Texas at Houston 44

45 Quasi 3D-Centrifuge Micro-Mechanics Parts 4 or 100mm Molded CD 45

46 Last time!!! Simulation Last (AGAIN) time!! 46

47 First test 47

48 Solutions Minimum requirement 1) Protein G = IgG immunoglobulins 2) Sample containing (or not) Salmonella 3) Washing (detergent) solution: rinse 48

49 Solution 1 & 2 49

50 Solution 3 50

51 Sealing troubles 51

52 Ideas for Sealing 52

53 DAVD-DOF design directional acceleration vector driven displacement of fluids 53

54 Macro-results H H QC A (1:1000) QC B (1:1000) H H H QC E2 (1:1000) H

55 Meaning TOFMS Spectrums Obtained Using the D irect Capture-In Situ Solubilization Technique For Purified Sub Strains of Salmonella (QC A, QC B and QC E2). All Sub-strains tested have a peak at a MW of 15, 270 including sub-strain E2. Only sub-strain A has an additional peak at 17, 110, whereas only sub-strain B has peaks at 16, 170 and 16, 470. Signal associated with each sample of Salmonella substrain detected using this approach is associated with a maximum theoretical number of 1600 captured bacteria. 55

56 The end Actual situation: waiting for the feedback from the measurements to be done at Houston to compare results and see whether or not the µ device works. Thank you! 56