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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 - PowerPoint PPT Presentation
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Device to apply a coating on tissue
samples for MALDI imaging
Holly LiskeLaura PiechuraKellen SheedyJenna Spaeth
Amy Harms, PhDUW Biotechnology Center
Brenda Ogle, PhDDepartment of Biomedical Engineering
Presentation Overview
Background of MALDI imaging Project motivation Problem statement Design criteria Design matrices Final design Budget evaluation Future work
MALDI-MSI
www.mc.vanderbilt.edu
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
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
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
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
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
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
Design Matrix Adjustment
Several variables controlled by many independent components
Sprayer Method of application Enclosure Orientation
Created a component matrix to decide final design
Component Matrix
VerticalHorizontalOrientation
Detached covering
Integrated covering
Fume hoodEnclosure
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
Ideas Feature
Criteria for ranking
Reliability Adjustability User interaction Cost Maintenance Ease of manufacture
Ranking System (1-5), highest = best
Mega MatrixSpraying Component
2.354.104.303.452.251.00Total
214310.05Ease of
Manufacture
243310.10Maintenance
315340.15Cost
354450.20User
Interaction
255410.25Adjustability
254310.25Reliability
Produce irrigation system
Pneumatic sprayer
Automatic Spray Gun
Nozzle-valve and
pressure vessel
NebulizerRankCriteria
Mega MatrixMethod of Application
3.252.851.00Total
130.05
Ease ofManufacture
330.10Maintenance
420.15Cost
430.20
User Interaction
330.25Adjustability
330.25
Reliability
Movable Plate Stationary Sprayer
Stationary Plate Movable Sprayer
RankCriteria
Mega MatrixEnclosure
2.452.91.551.00Total
4350.05Ease of
Manufacture
5410.10
Maintenance
3250.15
Cost
4410.20User
Interaction
2510.25
Adjustability
---0.25Reliability
Removable CoverIntegrated CoverFume HoodRankCriteria
Mega MatrixOrientation
1.25.51.00Total
--0.05Ease of
Manufacture
--0.10Maintenance
--0.15Cost
--0.20User
Interaction
--0.25
Adjustability
520.25
Reliability
Vertical SprayHorizontal
SprayRankCriteria
Final Design Model
1. Automatic Spray Gun
2. Conveyor
3. DC Timing Motor
4. Integrated Polyethylene Box
14
3
2
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)
Future Work
Final Design modifications Order materials Build prototype Test in client’s lab
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.
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