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Device to apply a coating on tissue samples for MALDI imaging

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. Presentation Overview. Background of MALDI imaging Project motivation

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Device to apply a coating on tissue samples for MALDI imaging

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  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 Valine Alanine, neuropeptide in crab brain m/z = 1474.6 Valine, m/z = 1502.7 Greg Barrett-Wilt, PhD., UW Biotechnology Center

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

  6. 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 Matrix Application 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. Objective 1: Simplify matrix application, facilitate MALDI Objective 2: Centralize a reliable tool at UW Biotechnology Center Implications for campus-wide, even global, research Project Motivation 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 Feature Ideas Sprayer Automatic Spray Gun Nozzle-valve with pressure vessel Nebulizer Pneumatic sprayer Produce irrigation system Method of Application Movable sprayer, Stationary plate Stationary sprayer, Movable plate Enclosure Fume hood Integrated covering Detached covering Orientation Horizontal Vertical

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

  13. Mega MatrixSpraying Component Criteria Rank Nebulizer Nozzle-valve and pressure vessel Automatic Spray Gun Pneumatic sprayer Produce irrigation system Reliability 0.25 1 3 4 5 2 Adjustability 0.25 1 4 5 5 2 User Interaction 0.20 5 4 4 5 3 Cost 0.15 4 3 5 1 3 Maintenance 0.10 1 3 3 4 2 Ease of Manufacture 0.05 1 3 4 1 2 Total 1.00 2.25 3.45 4.30 4.10 2.35

  14. Mega MatrixMethod of Application Criteria Rank Stationary Plate Movable Sprayer Movable Plate Stationary Sprayer Reliability 0.25 3 3 Adjustability 0.25 3 3 User Interaction 0.20 3 4 Cost 0.15 2 4 Maintenance 0.10 3 3 Ease of Manufacture 0.05 3 1 Total 1.00 2.85 3.25

  15. Mega MatrixEnclosure Criteria Rank Fume Hood Integrated Cover Removable Cover Reliability 0.25 - - - Adjustability 0.25 1 5 2 User Interaction 0.20 1 4 4 Cost 0.15 5 2 3 Maintenance 0.10 1 4 5 Ease of Manufacture 0.05 5 3 4 Total 1.00 1.55 2.9 2.45

  16. Mega MatrixOrientation Criteria Rank Horizontal Spray Vertical Spray Reliability 0.25 2 5 Adjustability 0.25 - - User Interaction 0.20 - - Cost 0.15 - - Maintenance 0.10 - - Ease of Manufacture 0.05 - - Total 1.00 .5 1.25

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

  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.” <http://www.astbury.leeds.ac.uk/> • Barrett-Wilt, Greg. University of Wisconsin Biotechnology Center. Personal interviews. • Caprioli Research Laboratory Center. Vanderbilt Medical Center. <http://www.mcvanderbilt.edu> • 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?

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