1. Device to apply a coating on tissue samples for MALDI imaging Holly Liske Laura Piechura Kellen Sheedy Jenna Spaeth Amy Harms, PhD UW Biotechnology Center Brenda Ogle, PhD Department of Biomedical Engineering

2. Presentation Overview Background of MALDI imaging Project motivation Problem statement Design criteria Design matrices Final design Budget evaluation Future work

3. MALDI-MSI www.mc.vanderbilt.edu

4. Data Acquisition from MALDI-MSI Alanine, neuropeptide in crab brain m/z = 1474.6 Valine, m/z = 1502.7 Alanine Valine Greg Barrett-Wilt, PhD., UW Biotechnology Center

5. Benefits of MALDI-MSI Analysis of entire sample in one reading Previous knowledge of molecular composition is not necessary Allows for investigation of disease formation, progression, and treatment www.maldi-msi.org

6. Matrix Application Matrix application is vital for quality image resolution Must contact sample as fine, liquid mist Current procedure involves manual application with airbrush 100µm raster step Greg Barrett-Wilt, PhD., UW Biotechnology Center

7. Problem Statement Sample preparation methods MALDI-MSI are difficult to control Requires accuracy and precision A device to apply a fine, uniform coating of light- absorbing compounds

8. Project Motivation Objective 1: Simplify matrix application, facilitate MALDI Objective 2: Centralize a reliable tool at UW Biotechnology Center Implications for campus-wide, even global, research www.buswire.ocr.wisc.edu

9. Design Criteria Spray an even coating of matrix over an 81cm x 123 cm tissue sample Adjustable spray aperture, air pressure and positioning of the plate and sprayer Enclosed in a casing and operable within a fume hood

10. Design Matrix Adjustment Several variables controlled by many independent components Sprayer Method of application Enclosure Orientation Created a component matrix to decide final design

11. Component Matrix VerticalHorizontal Orientation Detached covering Integrated covering Fume hood Enclosure Stationary sprayer, Movable plate Movable sprayer, Stationary plate Method of Application Produce irrigation system Pneumatic sprayer Nebulizer Nozzle- valve with pressure vessel Automatic Spray Gun Sprayer IdeasFeature

12. Criteria for ranking Reliability Adjustability User interaction Cost Maintenance Ease of manufacture Ranking System (1-5), highest = best

13. Mega Matrix Spraying Component 2.354.104.303.452.251.00Total 21431 0.05 Ease of Manufacture 24331 0.10 Maintenance 31534 0.15 Cost 35445 0.20 User Interaction 25541 0.25 Adjustability 25431 0.25 Reliability Produce irrigation system Pneumatic sprayer Automatic Spray Gun Nozzle-valve and pressure vessel NebulizerRankCriteria

14. Mega Matrix Method of Application 3.252.851.00Total 13 0.05 Ease of Manufacture 33 0.10 Maintenance 42 0.15 Cost 43 0.20 User Interaction 33 0.25 Adjustability 33 0.25 Reliability Movable Plate Stationary Sprayer Stationary Plate Movable Sprayer RankCriteria

15. Mega Matrix Enclosure 2.452.91.551.00Total 435 0.05 Ease of Manufacture 541 0.10 Maintenance 325 0.15 Cost 441 0.20 User Interaction 251 0.25 Adjustability --- 0.25 Reliability Removable CoverIntegrated CoverFume HoodRankCriteria

16. Mega Matrix Orientation 1.25.51.00Total -- 0.05 Ease of Manufacture -- 0.10 Maintenance -- 0.15 Cost -- 0.20 User Interaction -- 0.25 Adjustability 52 0.25 Reliability Vertical SprayHorizontal SprayRankCriteria

17. Final Design Model 1.Automatic Spray Gun 2.Conveyor 3.DC Timing Motor 4.Integrated Polyethylene Box 1 4 3 2

18. Budget Evaluation $300.00 budget limit Breakdown Air Spray Gun: $150.00 Motor/Conveyor: $45.00 Enclosure material: $45.00 Other small parts: $40.00 Total Estimate: $280.00 (under budget)

19. Future Work Final Design modifications Order materials Build prototype Test in client’s lab

20. References Ashcroft, Alison. University of Leeds Astbury Centre for Structural Molecular Biology. “An Introduction to Mass Spectrometry.” Barrett-Wilt, Greg. University of Wisconsin Biotechnology Center. Personal interviews. Caprioli Research Laboratory Center. Vanderbilt Medical Center. Khatib-Shahidi, Sheerin. “Direct Molecular Analysis of Whole-Body Animal Tissue Section by Imaging MALDI Mass Spectrometry.” Analytical Chemistry. 2006, 78(18) 6448-6456. Sugiura, Yuki. “Two-Step Matrix Application Technique to Improve Ionization Efficiency for MALDI-MSI.” Analytical Chemistry. 2006, 78(24) 8227-8235.

21. Any Questions?