FROM IMAGES TO ANSWERS Portfolio de Productos de Media Cybernetics Olympus Latin America Q2 2006

New Releases for 2006 AutoQuant X InVivo/InVitro 3 Image-Pro Plus 6 (Slightly Higher Price but Lots More Stuff) Scope-Pro 6 (Piezo Support and More…) Image-Pro Analyzer 6 (Single Seat, 3-Seat) Image-Pro Express 6 (Lower Price; More Features) IPE 3DS IPE-MC Scope-Pro Express 6 (Z and Shutter Control) Vertical Applications: IQ-Otolith & IPP-Otolith Macro EDF Analyzer IQbase 2

New Releases InVivo 3

Top Features in InVivo 3 1-Button Data Set Export to Image-Pro Plus 6.0 Yes Finally Windows XP x64 Support Large memory address handling 64-bit camera drivers from QImaging and Hamamatsu Variable Rate Time Lapse Acquisition Nyquist Calculation for Z-Stack Acquisition Multi-Camera Support Support for Up To Four Concurrent Cameras True Simultaneous Acquisition from Two Monochrome Photometrics or QImaging Cameras Updated Driver Support for Other Popular Cameras

Top Features in InVivo 3 FRET Acquisition Live Calibrated Ratio Display

Top Features in InVivo 3, part deux Stage Navigation Upgrades Cell Chambers and Multi-Well Plate templates

Top Features in InVivo 3, part troix New Microscope Control Dialog Illustrations for Leica Stands Support for DM6000, 5000, 4000 & DMI6000, 5000, 4000

InVivo Family Software Series InVivoLive Cell Imaging Solution for Calcium Ratios, FRET and Other Dynamic Processes ExpIOVital Systems Control and Measurement ImageRenamed “InVitro” at Lower Price CaptureDiscontinued

Photometrics Family of Monochrome Cameras (including MAC, but NOT AS FAST! [no circular buffer support]) QImaging Family of Cameras (including MAC) Some Hamamatsu Cameras (not including Mac) Some Stanford Photonics Cameras Digital Camera Support

InVivo Acquisition Camera Control Fully Automated Microscope Control Multi Color Acquisition Time Lapse, Burst Acquisition 3D, z-series, Burst 3D with piezo Tiling, Mosaic, Slide and Well Scanning FRET Acquisition Optical Splitter Acquisition

Multi Dimensional Acquisition

InVivo Analysis Live Ratio and Ca ++ Imaging Intensity Calibration (Including Grynkewicz) Vital Systems Control and Measurement Signals and Systems control Continuous Wavelength Imaging Devices

Live Cell Imaging

Pre-Configured Systems Automated Microscopes Digital Cameras Stage Automation Vital Systems

Live Cell Imaging Application Solutions with InVivo Bundles

Multi Color Time Lapse Acquisition Live Intensity and Ratio Measurements with Fast Acquisition using InVivo Advanced Ratiometric and Intensity-based Tracking in Image-Pro Plus

Time Lapse, Burst Acquisition and Advanced Tracking High Speed Acquisition for Small Region(s) Track Single or Multiple Moving Particles

3D Acquisition, Deconvolution and Measurement Multicolor Z and Burst Z Acquisition Multi Color Deconvolution 2D & 3D Blind Advanced 3D Analysis and Measurements

Multidimensional Acquisition and Analysis Count Size Extended Depth of Field

Signals and Actions Acquisition and Analysis

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

New Releases Image-Pro Plus 6

Top New Features in Image-Pro 6 Live Image Scaling for Low-Light Applications Live Data Collection and Charting XP x64 Support to Open Larger Image Sets Open GL 3D Viewer Direct Import of QED Files (File, Open) User-Defined Equation Editor in Data Collector Dynamically Linked Graphic Display in Data Collector Embedded I/O Control for Serial and Parallel Devices ICC Color Profiles

New Open GL 3D Viewer

More File Support

New Data Collector Tools

New Data Input/Output Tool

New Color Management Tool

Image-Pro 6 Enhancements Image-Pro Plus 6 Feature Enhancements: Updated Capture Dialog Scalable Calibration Markers User-Specific Menus “Dark Mode” for Low Light Laboratory Environments Automatic Measurement of >100,000 Objects (Memory-Dependent) Intelligent Macro Editor (not Intelli-Sense, but something much like it ;) ) 3D Convolution Filters More Extensive File Support (FV1000, JPEG2000, etc.) Export to Excel or Origin Generate Movies from Surface Plot 6-Month License for IQbase 2 (Optional Permanent License at Extra Cost)

Updated Capture Dialog

New Scalable Calibration

New Menu Editor

New Dark Mode

New 3D Convolution Filters

Top New Features in Scope-Pro 6 3-Point Predictive Focus for Wavy Samples Support for Simultaneous Stepper and Piezo Drives (X, Y, and Z) Calibrated Continuous Zoom Support for Olympus SZ-X Objective Paracentricity (X,Y) Offsets Analog/TTL Support for Shutters, Filter Wheels, AOTF’s and Monochromators Nyquist Calculator for Z-Axis Acquisition Support for the Olympus ZDC & Nikon Perfect Focus

Image-Pro Family Modules Automate Motorized Microscopes and Other Acquisition Devices A/D & D/A Acquisition & Support for External Devices SharpStack Plus includes: 3D Blind, 2D Blind deconvolution algorithms plus No Neighbor, Nearest Neighbor and Inverse Filter algorithms Turbo Scan High Speed Tiling Back as a Plug-In by Popular Request

QImaging* (Excellent) Photometrics* (Excellent) Leica DFC* (Excellent) Olympus DP50 / DP70 / DP30 (Excellent) Nikon DXM1200/F DS5 (Good) Hamamatsu (Good, 3 rd Party Driver) Cooke PCO (Good, 3 rd Party Driver) Diagnostic Instruments (Fair, 3 rd Party Driver) Lumenera (Fair, 3 rd Party Driver) Camera Support for Image-Pro Family * Image-Pro 6.X Support

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

New Releases IQbase 2

Top New Features of IQbase 2 IQbase 2 Features: Capture Module (Optional extra Cost) Support for Active Directory – Use Windows Identities for log-in and Security Levels Export Images and Data to PowerPoint Apply Measurements to Archived Images Compare Images Side-by-Side using Lightbox Tool Use 3D Scatterplots to Mine Data in Three Dimensions Speed Optimized for Analysis, Searching, Opening and Displaying New Manual Measurement Tools

IQbase 2 Powering Image Informatics Manages Images and Data Drives Discovery Facilitates Collaboration Easy to Use Works the Way You Do Grows with Your Organization (Scalable)

IQstudio Analytical Imaging Development Kit Rapid Development of Image Analysis Applications Develop Applications Using our Tools Add Image Analysis to Existing Products Standardize Image Analysis within the Organization Distribute Custom Applications with Run- Time Licensing

Deconvolution, 3D Rendering & Measurement

Deblurring vs Deconvolution Auto-Quant® Algorithms are Used in Sharpstack The No Neighbor & Nearest Neighbor Algorithms are Deblurring Tools (Intensities are Removed) The Inverse Filter Algorithm is a Haze Restoration Tool (Intensities are Re-Assigned) The Optional 2D Blind and 3D Blind Tools are Deconvolution Algorithms (Intensities are Re- Assigned) The Sharpstack Algorithms are not Appropriate for Confocal Images Because They Use Widefield PSFs

Deblurring vs Deconvolution

AutoQuant X

AQI X Advantage Sequence detection –Multi time point –Multi channel Multi-Processing, Multi-Threaded Best in Deconvolution –Only Blind Algorithm Commercially Available –Spherical Aberration Detection and Correction

Sequence Detection Loads Complex File Sequences Dynamically Changes to Accommodate Naming of Current File

Multi Time Point Load 3D Time Series Data Visualize, Deconvolve and Analyze as a Single Dataset

Multi Channel Load any Number of Channels Change Tints & Thresholds in Real-Time

Multi-Processing, Multi-Threading Each Operation Runs in its Own Thread Deconvolve, Visualize a Dataset and Queue Another Dataset at the Same Time Run Two Deconvolutions Simultaneously Status Display is a Dashboard for Activity Takes Advantage of Dual Processor Computers

Blind Deconvolution Blind Deconvolution Yields the Best Results with the Least Effort No Need to Acquire or Maintain a Library of Point Spread Functions Adaptive PSF Changes with the Microscope: –Aberrations, Thermal Expansion, Noise, Alignment, and Specimen are Corrected Maximum Likelihood Expectation Models Noise Instead of Smoothing it

Hierarchy of Algorithms 3D Blind Deconvolution – Premier 2D Blind Deconvolution – Live cell 3D Non-Blind - Faster for Derived PSFs 3D Non-Blind (Gold’s) – Fast but Noisy 3D Inverse Filter, Nearest/No Neighbor – Quick and Dirty

Spherical Aberration Common Problem with RI Mismatches or Thick Samples Detect Using Iterative MLE or Calibrate Pre-Bias PSF Improves with Blind Deconvolution Improves Signal Due to Smearing

Deconvolution of Widefield and Confocal Images Primer on The Growing Role of Deconvolution…

Why Use Deconvolution? To Improve Signal to Noise Ratio

What is Deconvolution? Widefield – Single Channel

Multiple Wavelengths - Widefield

Confocal Microscopy

2D Blind Deconvolution

2D Blind Deconvolution – TIRF

When to Use Deconvolution? In Widefield Fluorescent Microscopy –Improve contrast of 2D and 3D Images –Improve Resolution in Three Dimensions –Improve 3D Rendering and Visualization

When to Use Deconvolution? In Confocal Microscopy –Improve Signal to Noise Ratios –Increase Dynamic Range –Improve Image Grayscale Resolution –Increase Axial Resolution

When to Use Deconvolution? Documentation –Images and Projections will Show More Detail and Greater Contrast –Improvements can be Striking

When to Use Deconvolution? Analysis –Deconvolution Retains Quantitative Accuracy –Improves Contrast Facilitating Analysis Counting Tracking Area Measurements Intensity Measurements

Why Does This Work? The Microscope is a Well-Understood Optical System The Behavior of Light is Well-Modeled Persistence Why this works (1/5)

The Point Spread of a Microscope Every Microscope- Sample Combination has a Different Point Spread Function Every Element in the Optical Path Affects the Point Spread Function: –Scope –Objective –Cover slip –Sample

The Point Spread of a Microscope Compare a widefield PSF to a Confocal PSF

Looking Forward to Deconvolution YFP Labeled Motor Neurons (Mouse) Laura Baylor, Harvard University Laboratory of Dr. Jeff Lichtman Emissive Wavelength Emissive Wavelength Numerical Aperture Numerical Aperture Spherical Aberration Spherical Aberration

Non-Blind deconvolution estimates the original object from the measured PSF and the collected data (2D or 3D image). Non-Blind Deconvolution

Blind deconvolution separates the original object and the point spread from the collected data (2D or 3D image). Blind deconvolution

3D User Interface

3D User Tools Pointer to Toggle between Measurements & Volume Display Pan Tool to Adjust 3D Volume Position Home Position Set New Home Position Change Perspective (View) Display Box Around Volume Change View Y,Z or Z,X or Y,X Use Control Click for LR or UD, Click for UD, Alt Click to Toggle Between Pan and Pointer

3D User Options

3D Rendering with 3D Filters

3D Volume Measurements

3D Volume Measurements Over Time

3D Branch / End Point Measurements

Silver Spring, MD (Headquarters) Worldwide Offices in: Boston, Pittsburgh & San Diego Leiden, The Netherlands Goettingen, Germany Wokingham, United Kingdom Singapore, Republic of Singapore Tokyo, Japan Shanghai, China