1. FROM IMAGES TO ANSWERS July 2005 Silver Spring and San Diego Advanced Quantitative Image Analysis

2. Agenda Day One 9:00-10:00Advanced Microscopy Suite Setup 10:00-11:00Multi-Dimensional Acquisition 11:00-NoonMulti-Dimensional Analysis Noon-1:00Lunch (Catered) 1:00-2:00Simulated Live Cell Acquisition & Analysis 2:00-3:00Simulated Ratio Acquisition and Analysis 3:00-4:00Piezo Z-Axis Setup & Fast Acquisition 4:00-5:00Group Discussion 5:00PMAdjourn 6:00PMDinner at a Nearby Restaurant

3. Agenda Day Two 9:00-10:00Deconvolution of Widefield Images 10:00-Noon3D Rendering and Measurement of Image Stacks Noon-1:00Lunch (Catered) 1:00-2:003D Rendering and Measurement of Image Stacks 2:00-3:002D & 3D Tracking 3:00-4:00Using Workflow Toolbars to Automate Processes 4:00-5:00Group Discussion 5:00Adjourn 6:00Dinner at a Nearby Restaurant

4. Agenda Day Three 9:00-10:00Industry Best Practices with Macros 10:00-11:00Communicating with External Devices 11:00-NoonIntegrating Macros with Applications (Libraries and Resources) Noon-1:00PMLunch (Catered) 1:00-2:00Fun with Dimensioning Variables & Arrays; Building Loops 2:00-3:00Practical Examples of Macros in Practice 3:00-4:00Transitioning to Visual Basic for Applications in IQbase 4:00-5:00Group Discussion 5:00Adjourn

5. 10 Things I Learned Doing IA Demos “Tell Me a Little About Your Research” (Stall) “It’s Never Done That Before” (Surprise) “Do You Have Administrator Privileges?” (Stall) “It Looks Like a Hardware Problem” (S/W Guy) “It Looks Like a Software Problem” (H/W Guy) “We’re Bringing in a Factory Expert” (We’re Clueless) “More Pixels are Better” (Pixel Envy) “Nobody Uses that Technology Any More” (Feed Ego) “All Your Colleagues are Using This Product” (Intimidation) “I’m Bringing in Our Imaging Specialist” (I’m Clueless)

6. Advanced Microscopy Setup FROM IMAGES TO ANSWERS

7. 9-Step Career Program Follow these Steps for a Rewarding Career… Install Application Program(s) Install Capture Driver and Save Settings (VPF File) Calibrate Objective(s) Configure and Save Microscope Settings Configure AFA from Saved Settings Acquire Multi-Dimensional Images Get Bigger Grant to Buy More Stuff Win Nobel Prize Retire

8. Camera Setup Install Capture Driver from WEB Site (CDs are Always Out of Date)

9. Camera Setup Create Video Preference File (VPF)

10. Microscope Setup Start Scope-Pro Configurator Select Microscope and Devices

11. Microscope Setup Start Scope-Pro Configure Devices Save Common Settings Calibrate Objectives

12. Microscope Setup Calibrate Objectives

13. Microscope Setup Set up Stage- Pro

14. Microscope Setup Import Objective Calibrations

15. Microscope Setup

16. Set Z-Limits Just Above & Below the Focal Plane

17. Microscope Setup Configure AFA using Microscope & Capture Settings

18. FROM IMAGES TO ANSWERS Live Cell Imaging - Practical Issues Silver Spring & San Diego, June 2005

19. QED Setup Install Application Program* Select Capture Driver from Pull-Down List Calibrate Objective(s) Select Microscope Settings from Pull-Down List or Auto-Configure Acquire Multi-Dimensional Images * Check for Program Updates (Support, Update this Software)

20. QED Camera Setup

21. QED Spatial Calibration Setup

22. QED Parfocality & Paracentricty Setup

23. QED Microscope Setup

24. QED Fluorescence Acquisition Setup

25. Live Cell Practical Considerations When selecting which system to use for imaging living cells, one should consider three things: –Sensitivity of Detection –Speed of Acquisition –Viability of the Specimen See “Light Microscopy Techniques for Live Cell Imaging”, David J. Stephens et al, Science, Vol. 300, pp. 82-86, 4 April 2003

26. What Do I Need to Control? –Camera (Gain & Binning) –Speed of Acquisition (Exp. Time) –Dimensions of Acquisition (X,Y,Z, Time, Wavelength, Location) –Shutters and Illumination (High or Low Speed Shutter, Hg or Xe Lamp) –Sample Environment (Temperature, Gases, etc.)

27. Sensitivity of Detection

28. What I See and What I Want to See

29. Camera Control Resolution of Interest Gain Exposure Time Binning

30. Speed of Acquisition

31. Time Lapse Experiments Low- and High- Speed Acquisition Track Single or Multiple Moving Objects or Intensities

32. Time Lapse Parameters Region of Interest Frame Rate Binning Exposure Time

33. Control Dimensions Time XY Color 3D Fast and Slow Hardware

34. Limiting Cell Damage

35. Shutter and Illumination control Limit light exposure. Smart Shutter Control

36. Gain and Binning Control

37. Viability of the Sample

38. Control the Environment and Sample

39. Multi Dimensional Live Cell Acquisition

40. Acquire Time Lapse Time interval

41. Creating True Color Images Imaging Modes Menu Choose 2 or More Channels Merge Channels Select Multiple Parameters

42. Multi-Channel Time Lapse Intensity and Ratio Measurements with Fast Acquisition Advanced Ratiometric and Intensity based tracking Multi-Channel Ratio Experiments with Automated Microscopes and External Devices

43. XYZ Acquisition Multicolor Z or High-Speed Burst Acquisition XY multi point or area scans

44. Multi-Dimensional Live Cell Acquisition Count Size 3D and Extended Depth of Field

45. Multi Dimensional Live Cell Analysis

46. Extracting Multi Channel Information

47. Ratio Analysis Intensity and Ratio Measurements with Fast Acquisition Advanced Ratiometric and Intensity based tracking Multi-Channel Ratio Experiments with Automated Microscopes and External Devices

48. Ion Channel and Fret Imaging

49. Tracking of objects Automatic Tracking Correlation Tracking on pattern rather than intensity threshold >50 Morphological Measurements available including Intensity Tracking Smart Search Radius Detects Object Movement from Image Coherence Filter Accounts for Directional Movement Static or Dynamic Intensity Tracking

50. 3D Deconvolution No Neighbor, Nearest Neighbor Haze Removal and Inverse Filter Haze Restoration Algorithms 2D Blind and 3D Blind Deconvolution Algorithms These Algorithms are not Appropriate for Confocal Images

51. 3D Reconstruction and Measurements

52. Image Pro 3D Suite Automatic Volumetric Measurements Over Time 4D Tracking of objects!!

53. Image Pro 3D Suite Branch/End Point Measurements

54. Publishing and Archiving

55. Show and Tell

56. Adding VRML content to Powerpoint - Install Cortona Browser Install Cortona Browser ® from the Parallel Graphics website: –http://www.parallelgraphics.comhttp://www.parallelgraphics.com –Cortona Browser will install for one of any number of browsers on your PC Display VRML content in a PowerPoint presentation

57. Managing Images and data Manages Images and Data Drives Discovery Facilitates Collaboration Easy to Use Works the Way You Do Grows with Your Organization (Scalable)

58. Competitive Feature Comparison InVivo/ Analyzer 3D Most Comprehensive Package in the Industry; Only 2 Upgrades Possible (Sharpstack Plus & IO Pro); No Charge for Drivers MetaMorph Extra Fee for Camera drivers, Hardware Drivers, 2D & 3D Blind Deconvolution & 3D Simple PCI Extra Fee for Automated Image Capture, Image Processing and Analysis, Dynamic Intensity Analysis, Motion Tracking and Analysis IPLab Extra Fee for Ratio Plus, Fluorescence CV for Mac, MultiProbe for Win, Motion Control, Shutter & Filter Control OpenLab Extra Fee for 3D Module, Morphology Module, Ratio Module, FRET Module, Colocalization Module, Registration Module Intelligent Imaging Extra Fee for Ratio, TTL, 3D, 4D Tracking, Multiwell Plate Module, Statistics Package AxioVision Extra Fee for Z-Stack, Time Lapse, Tiling, Autofocus, Multichannel, Measurements, Imaging Plus

59. Competitive Feature Comparison Smart Camera Control YNNNNN Advanced Zoom Navigator YNNNNN Advanced Microscope Control YYYYYY Tiling, Mosaic, Slide and Well Scanning YYYYYY Multi-dimensional Acquisition YYYYYY Multi-Color Imaging YYYYYY Time Lapse Imaging and Burst Acquisition YYYYYY Burst Color 3D with Piezo Focus YYNYNN

60. Competitive Feature Comparison MCMM/MFIPLab3ICompixAxioV Advanced Morphometric Measurment, Analysis & Classification YYYYYY Colocalization, Translocation FISH, and Fluorescence YYYYYY Ratio Imaging YYYYYY FRET Analysis YYYYYY Cell Tracking YYNYYN Vital Systems Control & Measurement YYNYNN Signals and Systems Control YYNYNN Data Analysis, Recording, and Reporting YYYYYY 3D rendering and measurements YNNYNN 3D Time Lapse Analysis YNNNNN Analog Digital signals support YYNYYN Trend chart analysis Intensity tracking YYYYYN

61. With Similar Features, How to Choose? Price (Highest to Lowest) MM > Zeiss > 3I > MC > Compix > Scanalytics Support (Best to Worst) MC > 3I > Compix > Zeiss > Scanalytics > MM Installed Base (Largest to Smallest) MC >> MM > Zeiss > Compix > Scanalytics > 3I

62. www.mediacy.com Silver Spring, MD (Headquarters) Offices in: Boston, MA, Pittsburgh, PA & San Diego, CA Leiden, The Netherlands Goettingen, Germany Wokingham, United Kingdom Singapore, Republic of Singapore Tokyo, Japan Shanghai, China