CHROMACAL User Guide (v 1.1) User Guide

CHROMACAL User Guide (v 1.1) User Guide

User Guide Notice Hello and welcome to the User Guide for the Datacolor CHROMACAL Color Calibration System for Optical Microscopy, a cross-platform solution that transforms your raw brightfield images into high quality, colorstandardized images and preserves the scientific integrity of your research data.. The CHROMACAL System is currently designed for and compatible with U.S. versions of Windows and Mac OSX operating systems (see System Requirements). No assurances can be provided that the CHROMACAL System will operate effectively in other versions of such operating systems. This User Guide was developed based on CHROMACAL Image Calibration software version 1.5 and CHROMACAL Monitor Calibration software version 4.5.9. Datacolor reserves the right to make periodic improvements and changes to the product(s) and software described in this User Guide, and as such, certain information contained in this User Guide may not reflect the most current functionality and/or user interface for the latest release of these products(s) and software. Datacolor will endeavor to update this User Guide as changes to the underlying product(s) and software occurs, but no assurances can be made regarding when or if such updates will be made. All efforts are made to ensure the accuracy of the information presented in this User Guide. However, should any errors be detected, please notify Datacolor at feedback.chromacal@datacolor.com. An extensive collection of additional support material, including a collection of frequently asked questions (FAQs) is available at www.scientific.datacolor.com/support and www.scientific.datacolor.com/resources. To obtain information on CHROMACAL distributors or agents in your area, visit our website at http://scientific.datacolor.com/contact-us. Support Questions? If you need help with a Datacolor CHROMACAL product, please contact CHROMACAL customer support at info.chromacal@datacolor.com or call (800) 982-6499. 2014 Datacolor. All rights reserved. Datacolor is a registered trademark of Datacolor, Inc. CHROMACAL is a trademark of Datacolor, Inc. Windows is a registered trademark of Microsoft Corporation in the United States and/or other countries. Mac is a registered trademark of Apple Computer, Inc., used under license. CHROMACAL User Guide (v 1.1) i

Contents Welcome... 1 CHROMACAL Overview... 1 User Guide... 2 System Requirements... 2 CHROMACAL Product Family and Packaging Contents... 3 CHROMACAL Color Calibration System for Optical Microscopy... 4 CHROMACAL Color Calibration Slide for Brightfield Microscopy... 4 CHROMACAL Monitor Calibration Set... 5 Installation Process... 6 CHROMACAL Monitor Calibration Software Installation... 6 CHROMACAL Image Calibration Software Installation... 7 Free Mode for CHROMACAL Image Calibration Software... 10 Licensed Activation and Registration Process... 11 Quick Start Instructions... 17 Software Overview... 17 Before You Begin... 17 Using CHROMACAL... 18 Quick Start to Monitor Calibration... 18 Quick Start to Image Calibration... 19 CHROMACAL Image Features... 29 Feature Overview... 29 Main Window in CHROMACAL Image... 29 Uncalibrated Monitor Message... 30 Calibration Slide Serial Number... 30 Recording the Calibration Slide Serial Number in CHROMACAL... 31 Aligning the Color Calibration Matrix... 32 Use Cropping Tools for Color Matrix Orientation... 33 Outline the Matrix to Expand Size... 34 Diagnostic Measurements of Calibration Slide... 36 Matrix Alignment... 37 Linearity... 38 Exposure... 39 Tools for Calibrated Specimen Images... 41 Viewing Tools... 42 Contrast Slider... 43 Reference Standard Label... 43 View CHROMACAL Images... 44 Send Your Feedback... 46 Menu Bar Options... 47 File Menu... 47 Open Calibration Slide Image... 47 Open CHROMACAL Log Folder... 47 Open CHROMACAL Application Folder... 47 Load Measurement File from Disk... 47 Quit... 48 Edit Menu... 48 Preferences... 48 View Menu... 49 ChromaCal Images... 49 Usage Stats... 49 Tools Menu... 50 Batch Process... 50 Measurement File Database... 50 Show Linearity Plot... 50 Monitor Menu... 51 Calibrate... 51 Analyze Quality... 51 Help Menu... 53 Appendix... 55 Appendix A: Glossary... 55 CHROMACAL User Guide (v 1.1) ii

Brightfield Imaging... 55 Calibration Slide Image... 55 Good Microscopy Practices... 55 Measurement File... 55 Uncalibrated Monitor... 55 CHROMACAL User Guide (v 1.1) iii

Welcome CHROMACAL Overview In brightfield imaging, it is widely observed that colors from the same specimens appear differently between microscopes, cameras and monitors. Sometimes the variation occurs between imaging sessions even though the same equipment is used. Can you trust your results when your images lack color integrity? Introducing Datacolor CHROMACAL, a cross-platform solution that transforms your raw brightfield images into high quality, color-standardized images and preserves the scientific integrity of your research data. With CHROMACAL you simply achieve the best image the first time, all the time...and avoid the time and risk of compromised data associated with image-editing software like Photoshop. If the quality and integrity of your research results is important, CHROMACAL should be your new best practice. CHROMACAL is an innovative, first-of-its-kind solution that enables researchers, scientists and technicians to establish and preserve the color integrity of their digital images in transmitted brightfield microscopy. CHROMACAL delivers its solution using an integrated image calibration and monitor calibration system. CHROMACAL: Standardizes and preserves color reproduction Includes automatic white balancing and matches brightness of calibrated images Delivers consistent, reliable basis for evaluation, communication, documentation and publication Performs image and monitor calibration for consistency throughout the imaging process Compatible with most microscopes, scientific cameras and acquisition software The CHROMACAL System integrates three components to establish a color profile for your microscopy system: Proprietary software for image and monitor calibration Precision-engineered color calibration microscope slide (sold separately; re-order #10-2010) Computer monitor calibration sensor (colorimeter) CHROMACAL easily integrates into your microscopy workflow. Simply capture your specimen images and an image of the proprietary CHROMACAL Color Calibration slide. Launch the CHROMACAL Image Calibration software (referred within this User Guide as "CHROMACAL Image"), select the calibration image and apply the calibration to any or all of your specimen images from the same microscopy session. All original images are preserved, and CHROMACAL-calibrated images are saved separately. Also, communicate your images with confidence on CHROMACALcalibrated monitors to view colors as they were intended. The outcome is improved consistency of images and enhanced communication and collaboration for better decision making. CHROMACAL..empowering science with color integrity. CHROMACAL User Guide (v 1.1) 1

User Guide This User Guide generally documents the Windows version of the CHROMACAL software. The OSX version has identical functionality, but screens and keystrokes may vary slightly. Hyperlinks are included to cross-reference additional sections that provide further details on a feature or function. Quick Start Instructions. Provides basic instructions for using the CHROMACAL software. Details on Select CHROMACAL Image Features. Provides additional details regarding certain features within the CHROMACAL Image software. Menu Bar Options. Documents the functions of each Menu Bar option. Appendix. This section includes a glossary of key terms and concepts to help you get the best results from CHROMACAL. NOTES For your convenience, many hyperlinks are embedded into this User Guide to facilitate: Accessing additional resources on the web Sending emails to Datacolor for product support and feedback Cross-referencing additional details within the User Guide When using cross-reference hyperlinks on a Windows workstation, you will be able to return to the section from which you initiated the hyperlink by using the <alt><left arrow> keys in combination. System Requirements Hardware and software requirements for CHROMACAL include the following: Operating System Windows XP 32/64; Win7 32/64; Win8 32/64; Mac OSX 10.7 or higher Connectivity Monitor Resolution Video Card RAM Fixed Disk Image Capture Internet Connection USB connection (powered port for monitor calibration sensor) 1024 768 or higher (1200 800 or greater is recommended) 16-bit (24-bit recommended) 128MB available 100MB available Unaltered TIFF or JPEG images acquired with scientificgrade camera Required for maximum workflow efficiency CHROMACAL User Guide (v 1.1) 2

CHROMACAL Product Family and Packaging Contents The CHROMACAL product family includes: CHROMACAL Image Calibration software (referred to in this User Guide as "CHROMACAL Image") o o A license to install on one (1) workstation is included as part of the CHROMACAL Color Calibration System for Optical Microscopy (re-order #10-1010) Additional workstation activations (re-order #10-4010) can be purchased separately through the software or at www.scientific.datacolor.com/additional-activations. Your original CHROMACAL Image license code will be required. CHROMACAL Monitor Calibration software (referred to in this User Guide as "CHROMACAL Monitor") o o Included as part of the CHROMACAL Color Calibration System for Optical Microscopy (reorder #10-1010) along with a CHROMACAL sensor (colorimeter); a license to install on three (3) workstations is provided Additional workstation activations (re-order #10-4020) can be purchased separately through the software or at www.scientific.datacolor.com/additional-activations. Your original CHROMACAL Monitor license code will be required. CHROMACAL Color Calibration Slide for Brightfield Microscopy o Required to use CHROMACAL Image software o Sold separately (re-order #10-2010) CHROMACAL Monitor Calibration Set (re-order #10-3010) o o o o o Ideal for facilities that may not perform brightfield microscopy but still understand the need for reliable and consistent color rendering on their monitors Also applicable to facilities that may want multiple CHROMACAL sensors to facilitate efficient and frequent monitor calibrations CHROMACAL Monitor Calibration Set can function without requiring the purchase of the CHROMACAL Color Calibration System. The set includes CHROMACAL Monitor software and CHROMACAL sensor. A license to install on three (3) workstations is provided. Additional workstation activations (re-order #10-4020) can be purchased separately through the software or at www.scientific.datacolor.com/additional-activations. Your original CHROMACAL Monitor license code will be required. CHROMACAL User Guide (v 1.1) 3

CHROMACAL Color Calibration System for Optical Microscopy Re-order #10-1010 - case contents D B C A Case Contents: The CHROMACAL System case includes the following: A B C USB Drive (memory stick). The USB drive contains both the CHROMACAL Image and CHROMACAL Monitor software. Insert into a USB port on your computer to install the program. See Installation Process for instructions. Monitor Calibration Sensor. The sensor is used in conjunction with CHROMACAL Monitor software to calibrate a monitor. It is recommended that monitors be calibrated once prior to using CHROMACAL Image (and then re-calibrated approximately every 90 days). Sensor Stand. Holds the CHROMACAL sensor when not in use. D Packaging Inserts. Includes the following: o Overview and Warranty Guide o License and Installation Guide o Quick Start Guide Monitor Calibration o Quick Start Guide Image Calibration CHROMACAL Color Calibration Slide for Brightfield Microscopy Re-order #10-2010 - case contents Case Contents: The CHROMACAL slide case includes the following: Color Calibration Slide The CHROMACAL slide (depicted above) is a unique, precision-engineered tool used in conjunction with the CHROMACAL Color Calibration System and your microscope to establish a baseline color profile that is unique to your imaging system and imaging session. (Note: The CHROMACAL slide is purchased and packaged separately, but is required to use the CHROMACAL Color Calibration System for Optical Microscopy). CHROMACAL User Guide (v 1.1) 4

Slide Holder. Designed to protect and store your CHROMACAL slide when not in use. Multi-Fold Insert. This insert provides detailed information on the CHROMACAL slide, including configuration, features, care instructions, and limited warranty description. CHROMACAL Monitor Calibration Set Re-order #10-3010 - box contents Box Contents: The box containing the CHROMACAL Monitor Calibration Set includes the following: USB Drive (memory stick). The USB drive contains the CHROMACAL Monitor software. Insert into a USB port on your computer to install the program. See Installation Process for further instructions. Monitor Calibration Sensor. The sensor is used in conjunction with CHROMACAL Monitor software to calibrate a monitor. It is recommended that monitors be calibrated once prior to using CHROMACAL Image (and then re-calibrated approximately every 90 days). Sensor Stand. Holds the CHROMACAL sensor when not in use. Packaging INSERtS. Include the following: o o License Code, Warranty and Installation GuidE Quick Start Guide CHROMACAL User Guide (v 1.1) 5

Installation Process The CHROMACAL System includes two software applications: CHROMACAL Monitor Calibration and CHROMACAL Image Calibration. The applications are provided in both OSX and Windows versions. The software is stored on a USB drive included in the system case. The installation procedure is similar to other OSX and Windows-based products. After successful installation of both software applications, the icons below should display on your desktop. CHROMACAL Monitor Calibration CHROMACAL Image Calibration CHROMACAL Monitor Calibration Software Installation This section documents the installation on a Windows workstation. 1. Insert the USB drive (the memory stick supplied inside the system case) into a USB port on your Windows workstation. 2. Browse the USB drive contents to locate the Windows folder. 3. Select (by double-clicking) the Windows folder to see its contents. 4. Launch the file named ChromaCal Monitor Calibration Setup.exe (or similar). Follow the on-screen instructions. 5. Upon completion of the installation routine, remove the USB drive and store in the CHROMACAL System case. The USB drive is required every time you install CHROMACAL Monitor software on another workstation. This section documents the installation on Mac OSX workstation. 1. Insert the USB drive (the memory stick supplied inside the system case) into a USB port on your Mac workstation. It should open and appear as a window on your desktop. 2. Double-click on the "Macintosh" folder in that window to open the folder that contains the two Macintosh installers for CHROMACAL. 3. Launch the "ChromaCal Monitor Calibration Installer" application, and then follow the onscreen instructions. 4. Upon completion of the installation routine, remove the USB drive and store in the CHROMACAL System case. The USB drive is required every time you install CHROMACAL Monitor software on another workstation. You are now ready to launch the ChromaCal Monitor Calibration software. Click the applicable icon on your desktop to begin your ChromaCal Monitor experience! CHROMACAL User Guide (v 1.1) 6

CHROMACAL Image Calibration Software Installation This section documents the installation on a Windows workstation. 1. Insert the USB drive (the memory stick supplied inside the System case) into a USB port on your Windows workstation. 2. Browse the USB drive contents to locate the Windows folder. 3. Select (by double-clicking) the Windows folder to see its contents. 4. Launch the file named CHROMACAL Image Calibration Setup.exe (or similar) and then follow the on-screen instructions summarized below: a. Select the language that will be used for the installation screens and click OK. b. Review the Welcome screen and click Next. c. Review the License Agreement in its entirety. If you accept the terms and conditions of the License Agreement, click Yes. If you do not accept the terms and conditions, DO NOT PROCEED WITH INSTALLATION. Click No and the installation process will terminate and the installation screen will close. To continue the remainder of the installation instructions you must click Yes. CHROMACAL User Guide (v 1.1) 7

d. Review the default installation location for the program (i.e. destination directory). We recommend that you accept the default destination. To accept the default destination, click Next. To change the location, click Browse and select the desired destination directory. When you have identified the desired location, click Next. e. The Ready to Install screen gives you a final chance to change your setup selections. Click Back to make any necessary changes. When you are ready to proceed with installation, click Next. The installation will begin; upon completion, you will be notified on the screen. 5. Upon completion of the installation routine, remove the USB drive and store in the CHROMACAL System case. The USB drive is required every time you install CHROMACAL Image software on another workstation. CHROMACAL User Guide (v 1.1) 8

6. You are now ready to launch the CHROMACAL Image Calibration software. Click the applicable icon on your desktop to begin your CHROMACAL Image experience! This section documents the installation of CHROMACAL Image on Mac OSX workstation. 1. Insert the USB drive (the memory stick supplied inside the system case) into a USB port on your Mac workstation. It should open and appear as a window on your desktop. 2. Double-click on the "Macintosh" folder in that window to open the folder that contains the two Macintosh installers for CHROMACAL. 3. Launch the "ChromaCal Image Calibration Installer" application, and then follow the onscreen instructions. 4. Upon completion of the installation routine, remove the USB drive and store in the CHROMACAL System case. The USB drive is required every time you install CHROMACAL Image software on another workstation. 5. You are now ready to launch the CHROMACAL Image Calibration software. Click the applicable icon on your desktop to begin your CHROMACAL Image experience! CHROMACAL User Guide (v 1.1) 9

Free Mode for CHROMACAL Image Calibration Software The CHROMACAL Image software offers a "free version" option. This "free mode" allows you to install / activate the software on an unlimited number of workstations without a license code. Within this free mode, various features are available so that you can experience CHROMACAL color calibration in action, including: Perform a diagnostic check on the linearity of your imaging system by simply opening a CHROMACAL calibration slide image (taken at any imaging workstation) in the CHROMACAL Image software; Perform and preview CHROMACAL color calibration of your specimen images in Single Preview mode (Note: to save the images, a full activation is required); and View any previously saved ChromaCal-calibrated images, whether created by you or any of your colleagues or associates. The first time you launch the CHROMACAL Image software, a CHROMACAL Image Calibration Activation screen displays. To use the free version, click Use Free Version. If, after using the free mode, you want to experience the full features of CHROMACAL Image software, click Activate All Features and you will go to the Licensed Activation screen. CHROMACAL User Guide (v 1.1) 10

Licensed Activation and Registration Process When you first launch CHROMACAL Monitor Calibration software, you will be required to activate and register the software. To activate, enter the unique license code printed within your packaging inserts when prompted (see sample below). When you first launch CHROMACAL Image Calibration software, you have the option of running the software in "Free Mode" (no license code required) or the full version (license code required). When activation with a license code is necessary, enter the unique license code printed within your packaging inserts when prompted (see sample below). NOTES A separate activation and registration process is required for each CHROMACAL workstation unless you are using the "Free Mode". Also, separate license codes are assigned to each program (CHROMACAL Monitor Calibration and CHROMACAL Image Calibration). The procedure is identical, and detailed within this section. License and Installation Guide (CHROMACAL System case) License Code, Warranty and Installation Guide (CHROMACAL Monitor Calibration Set box) IMAGE CALIBRATION SOFTWARE LICENSE CODE # XXXXXX-XXXXXX-XXXXXX MONITOR CALIBRATION SOFTWARE LICENSE CODE # XXXXXX-XXXXXX-XXXXXX MONITOR CALIBRATION SOFTWARE LICENSE CODE # XXXXXX-XXXXXX-XXXXXX CHROMACAL User Guide (v 1.1) 11

NOTES CHROMACAL Image Calibration activation screens are shown below for demonstration purposes. The process is similar for the CHROMACAL Monitor Calibration software, but those screens simply identify a different software name. A "Free Version" is not available for the CHROMACAL Monitor Calibration software. Online Activation (Licensing) Process If you have an internet connection on your workstation (recommended environment for activation / licensing), the following process is applicable to your software activation process. If you do not have an internet connect on your workstation, go to Off-Line Activation (Licensing) Procedure. 1. The first time you launch a CHROMACAL program (or after clicking Activate All Features in the free mode of CHROMACAL Image software), a CHROMACAL Activation screen displays. To use the free version, click Use Free Version. To activate the full features of CHROMACAL, enter the 18-digit license code provided within the packaging insert. Do not enter the dashes as part of the license code. After entering the 18-digit code, click OK. 2. The program will provide a confirm message that will vary based on your choice above. If free version... If full version / features... 3. Click OK. If you are using the full version, you will proceed to the Registration screen. If an error message is shown instead, follow the on-screen instructions to resolve. CHROMACAL User Guide (v 1.1) 12

Off-Line Activation (Licensing) Procedure The following process is used to activate the software when there is no internet connection on your workstation 1. The first time you launch a CHROMACAL program, a CHROMACAL Offline Licensing screen displays. To use the free version, click Use Free Version and begin your CHROMACAL experience. To use the full features of CHROMACAL, click the "Begin the licensing process..." button. 2. Enter the 18-digit license code provided within the packaging insert as described earlier. Do not enter the dashes as part of the license code. CHROMACAL User Guide (v 1.1) 13

3. After entering the 18-digit code, click Generate Request. Additional instructions will display on the left side of the screen. 4. On the right side, an Activation Request code is shown and highlighted. Copy this code onto a USB memory stick or other portable storage device. It will be easiest to use the copy function, as it is a long string of characters. Once copied onto a portable device, click Finish. 5. Go to a workstation with an internet connection and send an email to Datacolor (feedback.chromacal@datacolor.com) with the Activation Request code from step #4 pasted into the email. 6. After receiving your email, Datacolor will return an Activation Certificate code to the same email address you used in step #5. Copy this code onto a USB memory stick or similar portable storage device. 7. With the USB memory stick from step #6, return to the workstation where you are trying to activate the CHROMACAL program. Insert the USB memory stick containing the Activation Certificate code. Relaunch the CHROMACAL software (if you exited the software). 8. The CHROMACAL Offline Licensing screen should display. Click the "Complete the licensing process..." button. CHROMACAL User Guide (v 1.1) 14

9. Enter the Activation Certificate code from step #6 into the labeled area on the right side of the screen. It will be easiest to copy the code from your USB memory stick (or other portable storage device) and paste the code into the Activation Certificate box. Then, click Activate. Your activation is now complete and you will proceed to the Registration screen. If an error message is shown instead, follow the on-screen instructions to resolve. NOTES It is best to use the on-line activation process. If you are able to restore the internet connection on your workstation, it will be easiest to do so and continue with on-line registration. Offline activation can be cumbersome and is intended only for those systems that do not have internet access. CHROMACAL regularly re-verifies your license code. If you have an internet connection, this occurs in the background and no input from you is required. If you do not have an internet connection, you will be periodically prompted to follow the Off-Line Activation Process previously described. We apologize in advance for any inconvenience, but it is necessary to protect the product from unauthorized use. CHROMACAL User Guide (v 1.1) 15

Registration Process After a successful licensed activation of the CHROMACAL program, you are requested to complete the following registration screen. The information assists Datacolor in continuously improving the performance and features of CHROMACAL, and to assist in Customer Support. 1. Completion of the registration information is only required once for each CHROMACAL program installation. All fields labeled in yellow that include an asterisk ("*") must be completed. 2. We encourage you to " " the first three questions at the bottom of the screen. This will greatly assist in our ongoing development of CHROMACAL. Thank you in advance for your cooperation! a. You can change your registration information at any time in the Help / Registration function accessible from the Menu Bar. 3. When all required information is entered, the OK button will activate. Click OK to finish and launch your CHROMACAL experience! CHROMACAL User Guide (v 1.1) 16

Quick Start Instructions Software Overview Introducing Datacolor CHROMACAL, a cross-platform solution that transforms your raw brightfield images into high quality, color-standardized images and preserves the scientific integrity of your research data. With CHROMACAL you simply achieve the best image the first time, all the time...and avoid the time and risk of compromised data associated with image-editing software like Photoshop. If the quality and integrity of your research results is important, CHROMACAL should be your new best practice. CHROMACAL: Standardizes and preserves color reproduction Includes automatic white balancing and matches brightness of calibrated images Delivers consistent, reliable basis for evaluation, communication, documentation and publication Performs image and monitor calibration for consistency throughout the imaging process Compatible with most microscopes, scientific cameras and acquisition software The CHROMACAL System integrates three components to establish a color profile for your microscopy system: Proprietary software for image and monitor calibration Precision-engineered color calibration microscope slide (sold separately; re-order #10-2010) Computer monitor calibration sensor (colorimeter) Before You Begin CHROMACAL easily integrates into your microscopy workflow. Simply capture your specimen images and an image of the proprietary CHROMACAL Color Calibration slide. Launch the CHROMACAL Image Calibration software (referred within this User Guide as "CHROMACAL Image"), select the calibration image and apply the calibration to any or all of your specimen images from the same microscopy session. All original images are preserved, and CHROMACAL-calibrated images are saved separately. Also, communicate your images with confidence on CHROMACALcalibrated monitors to view colors as they were intended. The outcome is improved consistency of images and enhanced communication and collaboration for better decision making. A very important part in successfully color-calibrating your images will be to capture your specimen slides and the CHROMACAL Color Calibration Slide using good microscopy practices. Recommended guidelines are available at www.scientific.datacolor.com/resources/microscopy-practices. We also recommend that you read the Frequently Asked Questions (FAQs) at www.scientific.datacolor.com/support/faq. This Q&A format provides quick and useful insights that should further assist you in obtaining a successful CHROMACAL experience. CHROMACAL User Guide (v 1.1) 17

Using CHROMACAL Quick Start to Monitor Calibration Invest the time to calibrate your monitor! From start to finish, monitor calibration should take less than 10 minutes. BEST PRACTICES Color-calibrating every monitor used for displaying digital images is a critical element to achieving an optimized viewing experience. All monitors used for the capture, evaluation and communication processes (at your workstation, within your facility, and among your collaborators) should be colorcalibrated with CHROMACAL Monitor Calibration software. This will tune your displays to a reference standard to bring color consistency across monitors. After calibration, images can be viewed and evaluated with greater confidence. The monitor calibration process is recommended after you initially install the software, and then should be updated periodically. We suggest every 90 days, but the frequency should be determined based on your environment and requirements. A reminder period can be set within the software. Step 1: Install the software (if you have not already done so) See CHROMACAL Monitor Calibration Installation for further instructions on software installation. Step 2: Start-up environment Your monitor should be on for at least 30 minutes before you begin the calibration process. Be sure there is no direct light shining on your monitor. Reset the monitor controls to the factory default settings, if available. Step 3: Launch the ChromaCal Monitor Calibration application When you use the application for the first time, you will be required to activate / register the software by providing the unique license code provided in the packaging. See Activation and Registration Process for further instructions. Continue by carefully reading and following the application s directions and prompts. These will guide you to ensure successful calibration. Step 4: Plug in the sensor only when prompted by the software When prompted, plug the sensor into a powered USB port on your workstation. Hang the sensor over your monitor (see illustration below) and position the sensor within the frame outline displayed on the screen. Slide the round counterweight on the USB cord so that the sensor stays in the correct location. When the calibration routine has completed its use of the sensor, you will be prompted to remove it from your monitor. CHROMACAL User Guide (v 1.1) 18

Step 5: Complete the calibration process Your CHROMACAL Monitor Calibration software will lead you through the remainder of the calibration process. The calibration procedure creates a custom display profile which your operating system and color-management applications (such as CHROMACAL Image Calibration software) will use to correct your displayed colors. Step 6: See the CHROMACAL difference When the calibration process is complete, be sure to view the color improvement by using the SpyderProof feature to see calibrated and uncalibrated sample images. For additional information on the CHROMACAL Monitor Calibration software, select Help/ ChromaCal Monitor Calibration Help from the main menu bar. Quick Start to Image Calibration CHROMACAL Image Calibration easily integrates into your microscopy workflow. Simply capture your specimen images and an image of the proprietary CHROMACAL Color Calibration slide. Launch the CHROMACAL Image Calibration software (referred within this User Guide as "CHROMACAL Image"), select the calibration image and apply the calibration to any or all of your specimen images from the same microscopy session. All original images are preserved, and CHROMACALcalibrated images are saved separately. Separately, communicate your images with confidence on CHROMACAL-calibrated monitors to view colors as they were intended. The CHROMACAL outcome is improved consistency of images and enhanced communication and collaboration for better decision making. CHROMACAL: Standardizes and preserves color reproduction Includes automatic white balancing and matches brightness of calibrated images You should strive to capture your specimen slides and the CHROMACAL Color Calibration Slide using good microscopy practices. Recommended guidelines are available at http://www.scientific.datacolor.com/resources/microscopy-practices. We also recommend that you read the Frequently Asked Questions (FAQs) at http://www.scientific.datacolor.com/support/faq. This Q&A format provides quick and useful insights that should further assist you in obtaining a successful CHROMACAL experience. The steps for image calibration are (and details follow): Step 1: Begin your microscopy session; check microscope and camera settings Step 2: Capture your ChromaCal Calibration Slide image Step 3: Check for Linearity and Correct Exposure Step 4: Capture your specimen slide images Step 5: Calibrate your specimen slide images CHROMACAL User Guide (v 1.1) 19

Step 1: Begin your microscopy session; check microscope and camera settings a. Conduct your microscopy session using good microscopy practices. Start by establishing Koehler, setting illumination and exposure, and white balancing. o Additional guidance on good microscopy practices is available at http://www.scientific.datacolor.com/resources/microscopy-practices. b. Adjust any settings within your image capture software that remove the linearity of captured images. For example: o any setting for "color correction" should be turned off o any gamma setting should be set to 1 NOTE Linearity is important because it keeps image colors at their original tones, as close to what is seen in the specimen as possible. To accomplish that, cameras must retain not just the darker tones, but the relationship or linearity of tones from dark to light. In other words, the darker colors cannot be made brighter without also brightening the lighter colors. For color calibration to be effective, the original images must be linear (see Step #2 and #3 to verify linearity). Step 2: Capture your CHROMACAL Calibration Slide Image a. After your initial set-up, place the CHROMACAL Color Calibration Slide (purchased separately) onto the microscope stage and capture an image of the larger 20-color matrix. Although the matrix area on the slide contains two separate matrices, we recommend you use the larger matrix which is generally visible at magnifications between 4x and 40x. NOTES DO NOT use an oil immersion lens with the calibration slide. When acquiring a calibration slide image, capture either the large or small color matrix, but not both. Generally, we recommend use of the larger matrix regardless of the magnification used to capture your specimen images. However, if optimal precision in the color calibration is required (for example, when the calibrated images will be used for quantitative analysis), we recommend you capture calibration side images at each of the magnifications used to capture your specimen images. The calibration slide image can be acquired at any time during your microscopy session, although we suggest an early capture to check the linearity of your imaging system. Capture the color matrix that best fits within the field of view of the objective on the microscope. If the microscope objective is currently on 10X-40X, use the larger matrix If the microscope objective is currently 40X and above (and is not an oil or water immersion lens), capture the smaller matrix. CHROMACAL User Guide (v 1.1) 20

When the calibration slide image is exposed correctly, the image will appear dark and certain patches may not be visible. This is normal. Examples of proper and incorrect exposure of the calibration slide are below. Proper Exposure Incorrect Exposure Y Y o u You can set the correct exposure in any of the following ways: i. If you are using auto-exposure, allow the camera to set exposure, but only if the calibration slide image appears dark and the brightest white circle appears slightly gray. If the image is over-brightened, turn off auto-exposure and set exposure manually according to ii or iii below; ii. Use a histogram in your camera software to be certain that the brightest part of the histogram display is within the dynamic range of the camera as you reduce or increase your exposure; or iii. Set by eye; reduce exposure until the brightest white circle becomes slightly gray and the second brightest white circle appears noticeably darker in comparison. b. Save the calibration slide image as a TIFF or JPEG file. BEST PRACTICES For easy access, save the calibration slide image in the same folder as your specimen images from the same microscopy session. We also recommend that you include the calibration slide serial number (the 7-digit number located in the upper right-hand corner of the calibration slide) as part of the file name you assign to the calibration slide image. This method will help in Step 3b. CHROMACAL User Guide (v 1.1) 21

Step 3: Check for Linearity and Correct Exposure a. Launch the CHROMACAL Image Calibration software. NOTES If your monitor has not been calibrated with CHROMACAL Monitor software, an "Uncalibrated Monitor" (or similar) message will appear on the above screen. See Uncalibrated Monitor for further details. b. Click Open Calibration Slide Image and locate the file of the calibration slide image created during the microscopy session (see Step 2b above). Highlight the desired file and click Open. CHROMACAL User Guide (v 1.1) 22

If this image file was never previously used in CHROMACAL Image software on your workstation, the program will prompt you for the serial number of the calibration slide. When prompted, select from the drop-down list (or get another if not listed) the 7-digit serial number associated with the calibration slide used to create the calibration slide image. The slide serial number is located in the upper right-hand corner of the CHROMACAL slide. If the slide serial number is not in the drop-down list, click "Get another". See Calibration Slide Serial Number for further information, if needed. c. Align the calibration matrix. Check the position of the color matrix. Use the Flip or Rotate tools if any adjustments are needed. o o It is critical that the white circle be located in the lower-right hand corner of the matrix The image should generally be aligned horizontally See Aligning the Color Calibration Matrix for further information. NOTES The colors displayed in the corners of the Calibration Profile window below are reminders of how the matrix should be oriented. For illustrative purposes, the sample screen below shows both the large and small matrix. When acquiring a calibration slide image, you will typically capture either the large or small color matrix, but not both. (Note: the above matrix is NOT aligned) CHROMACAL User Guide (v 1.1) 23

d. With the Magnifier tool selected, press and hold your mouse button starting near the top left-hand corner of the 20-color matrix (green circle) and drag the selection rectangle to the lower right-hand corner of the 20-color matrix (white circle). Upon releasing the mouse button, the color matrix will expand to fill the window. (Note: the matrix above is still NOT properly aligned) e. When Step 3d is completed, a blue square should fit completely within each color circle. The exact location within each circle is not critical. o o If the squares are orange, then your matrix is NOT aligned properly. If any adjustments are needed, re-align the squares by using the location adjustments displayed as your mouse hovers over each square. The adjustments vary depending on the square location. CHROMACAL User Guide (v 1.1) 24

(Note: the matrix above is properly aligned) NOTES In the lower left-hand corner of the screen are diagnostics related to the calibration slide image. See Matrix Alignment, Linearity and Exposure for further explanations. As noted in Step 2a, it is critical that the white circle be located in the lower right-hand corner of the matrix. Use the Flip and Rotate tools to make this adjustment if needed. f. Check the diagnostic readings. You want to achieve the following diagnostic results before proceeding to (i) capture your specimen images and (ii) color calibrate your specimen images. Matrix Alignment = "Good" (if not, realign the color matrix or the squares that overlay the matrix) Linearity = "Good" or "Within Tolerance" (if not, check camera software settings and repeat Step 2; pay particular attention to guidance provided in Step 1) Exposure = "Good" (if not, repeat Step 2 and pay particular attention to the exposure guidance provided). When the above results are achieved, proceed to Step 4. Step 4: Capture your specimen slide images a. Acquire specimen images as usual. b. Save your specimen images as TIFF or JPEG files. Step 5: Calibrate your specimen slide images a. Return to the CHROMACAL Image software. If you closed the CHROMACAL Image software after Step 3, then re-launch the application and follow Steps 3b through 3f. b. Once the recommended diagnostic readings (see Step 3f) are achieved, then: Open Single Image to color-calibrate one specimen image at a time (continue to Step 5c) or Batch Multiple Images to color-calibrate a batch of specimen images (continue to Step 5d). CHROMACAL User Guide (v 1.1) 25

c. Click Open Single Image to calibrate one image at a time. Locate and select the file with the specimen image that you want to calibrate. The ChromaCal Image Calibration Preview screen will appear. Your original image will be displayed on the left and the CHROMACAL color-calibrated image on the right. Use the Magnify and Hand Tools on the left of the screen for various viewing options. See Viewing Tools for further details Adjustments to the calibrated image are available for Contrast and for the Reference Standard Label. See Contrast Slider and Reference Standard Label for further details. When you are finished, click Save Calibrated Image to save the image. CHROMACAL User Guide (v 1.1) 26

NOTES Each calibrated image will be saved into a separate file (naming convention = original image file name PLUS the word CHROMACAL-calibrated appended to the end). No changes are made to the file containing the original image. The file type (TIFF or JPEG) for the calibrated image will be the same as the original image file. If additional images require calibration, click Open Image; otherwise, click Finished. d. Click Batch Multiple Images to color-calibrate an entire folder of specimen images. Click Select Source Images Folder to identify the folder location of the non-calibrated images. Once a source folder is selected, the file names of all uncalibrated images in that folder will be displayed on the left side of the screen. You can then highlight any file name and two images will be shown: the original image on the left and the CHROMACAL colorcalibrated image on the right. CHROMACAL User Guide (v 1.1) 27

After reviewing the image(s) and/or changing contrast (or other parameters) for any image, click Select Destination Images Folder to specify where the color-calibrated images should be saved. (See Viewing Tools, Contrast Slider and Reference Standard Label for additional viewing and parameter adjustments). Click Process Images. All images with a " " next to the file name will be processed and CHROMACAL color-calibrated versions of each of these images will be saved into the destination folder. When processing is done, a confirmation screen will display. Click OK to clear the message. When done with the Batch Processing function, click Finished. NOTES To select or deselect a file, click the checkbox next to the file name. To deselect all files, click the icon at the top of the column. To re-select all files, click the icon at the top of the column. Each calibrated image will be saved into a separate file (naming convention = original image file name PLUS the word ChromaCal-calibrated appended to the end). No changes are made to the file containing the original image. The file type (TIFF or JPEG) for the calibrated image will be the same as the original image file. CHROMACAL User Guide (v 1.1) 28

CHROMACAL Image Features Feature Overview This section provides additional details on select features and functions with the CHROMACAL Image Calibration software. Main Window in CHROMACAL Image 1. After launch the CHROMACAL Image Calibration software, the following window displays: Menu Bar Calibration Profile Window Menu Bar. ALL Program features can be accessed from the Menu Bar at the top of the screen. Open Calibration Slide Image. Opens the calibration slide file and begins the color matrix alignment. Once you align the color matrix, you can color-calibrate all specimen images collected during the same microscopy session. Calibration Profile window. Location of the calibration slide image after you open it in CHROMACAL. Send your Feedback. At any time, you can send an email to CHROMACAL customer support regarding the software. This includes general questions, product suggestions and troubleshooting information. CHROMACAL User Guide (v 1.1) 29

Uncalibrated Monitor Message When launching CHROMACAL Image software, the Main screen displays. During this process, the program checks whether your monitor is calibrated. If your monitor has not been calibrated, or your monitor has not been calibrated with a CHROMACAL sensor, you will be notified with orange text above the Calibration Profile window similar to the screen shown below. BEST PRACTICES Although you can continue with image calibration, it is highly recommended that you calibrate your monitor with CHROMACAL. Without such calibration, you cannot be assured that the color rendered on your monitor will be optimal and standardized. See CHROMACAL Monitor Calibration for instructions on monitor calibration. Calibration Slide Serial Number Every CHROMACAL Color Calibration slide has a matching Measurement File that contains the unique color profile of the slide. CHROMACAL software cannot determine the measurement file associated with a calibration slide image until you identify the serial number. It is critical that you assign the serial number correctly. BEST PRACTICES To simplify the identification of a serial number, it is best to include the serial number within the image file name assigned during your microscopy session. When this practice is followed, you can easily reference the serial number during your CHROMACAL Image session. CHROMACAL User Guide (v 1.1) 30

Recording the Calibration Slide Serial Number in CHROMACAL File name previously assigned to calibration image file 1. If you are using a calibration slide image file for the first time, you will be prompted to enter the 7-digit serial number of the associated slide. Select the correct number from the dropdown list (or get another if not listed). The slide serial number is located in the upper right-hand corner of the CHROMACAL Calibration slide. NOTES The drop-down list is populated with the contents of the CHROMACAL Calibration slide serial numbers you have used in the past. Any serial number you recorded previously in CHROMACAL will be shown on the drop-down list. If you followed a "Best Practice", the calibration image file name will contain the serial number and it will be visible and easy for you to reference! If the wrong calibration slide is assigned to the image file, you can delete this assignment and assign the correct slide image. See Measurement File Database to delete the information. This procedure assumes the workstation you are using has an internet connection. After selecting a serial number from the drop-down list, the software will immediately display the Main window and you can begin to align the matrix. 2. If you clicked "Get another", the following window will appear. File name previously assigned to calibration image file 3. Enter the 7-digit serial number of the associated calibration slide image and click Search. The slide serial number is located in the upper right-hand corner of the CHROMACAL Calibration slide. 4. If the serial number is valid, the program will display additional information about that slide. CHROMACAL User Guide (v 1.1) 31

5. Click Import and the Measurement File will be downloaded from the Datacolor Cloud Server. A confirmation message will displayed to confirm a successful download of the Measurement File to a local database. If the serial number does not exist, an error message will be displayed. Double-check the serial number and carefully re-enter. 6. Click OK to exit the procedure. Aligning the Color Calibration Matrix After opening the color calibration image, the Main screen below will be displayed and the unaligned calibration matrix is shown in the Calibration Profile window. As a reference to the alignment of the color matrix, the corners of the Calibration Profile window display the colors that should appear on each corner of the calibration slide image. The reference corners are highlighted below with arrows. CHROMACAL User Guide (v 1.1) 32

NOTES When acquiring a calibration slide image, capture either the large or small color matrix, but not both. Both 20-color matrices are shown above, just for illustration purposes. The calibration slide image can be acquired at any time during your microscopy session, although we suggest an early capture to check the linearity of your imaging system. Capture the color matrix that best fits within the field of view of the object on the microscope. If the microscope objective is currently on 10X-40X, use the larger matrix If the microscope objective is currently 40X and above (and is not an oil or water immersion lens), capture the smaller matrix. Generally, we recommend use of the larger matrix regardless of the magnification used to capture your specimen images. However, if optimal precision in the color calibration is required (for example, when the calibrated images will be used for quantitative analysis), we recommend you capture calibration side images at each of the magnifications used to capture your specimen image. The colors displayed in the corners of the Calibration Profile window are reminders of how the matrix should be oriented. The following 3 steps are the quickest way to achieve proper alignment: 1. The white circle must be located in the lower right-hand corner of the Calibration Profile window. Use Cropping Tools (Flip Tools) to correct. 2. The color matrix should be aligned horizontally. Use Cropping Tools (Rotate Tools) to correct. 3. A blue rectangle must be fully enclosed within each color circle. Outline the matrix and then move the rectangles to finalize alignment. Use Cropping Tools for Color Matrix Orientation Magnifier Tool (default tool). Use to magnify the color matrix (or <shift>click to shrink). The cursor will appear as a "+" symbol (or "-" symbol if in shrink mode) in the Profile window. Hand Tool. Click to activate. The cursor will change to a "hand" symbol when you drag it into the Profile window. Click and hold the mouse button to move the entire color matrix around the display window. Flip Tools. Click the horizontal icon to flip the color matrix horizontally. Click the vertical icon to flip the color matrix vertically. Rotate Tools. Click the counterclockwise icon to rotate the image counterclockwise. Click the clockwise icon to rotate the color matrix clockwise. The degree of rotation can be controlled as follows: Click icon: moves in 0.5 degree increments Shift+Click icon: moves in 15 degree increments Shift+Ctrl+Click icon: moves in 90 degree increments Reset. Return the color matrix to its original size and orientation CHROMACAL User Guide (v 1.1) 33

Outline the Matrix to Expand Size 1. With the Magnifier Tool selected, press and hold the mouse button starting at near the top left-hand corner of the 20-color matrix (green circle) and drag the selection rectangle to the lower right-hand corner of the 20-color matrix (white circle). 2. Upon releasing the mouse button, the color matrix will expand to fill the window. (Note: the above matrix is still NOT completely aligned) CHROMACAL User Guide (v 1.1) 34

3. A blue rectangle must be completely contained within each color circle. The exact location within each circle is not critical. If any adjustments are needed, re-align the squares. When you hover the mouse over each square, adjustments are shown. The adjustments vary depending on the square location. Below is an illustration of a properly aligned color matrix. (Note: the above matrix is properly aligned) As noted previously, the location of the white circle in the lower right-hand corner of the matrix is critical. CHROMACAL User Guide (v 1.1) 35

Diagnostic Measurements of Calibration Slide The CHROMACAL color calibration slide captures a profile of your imaging system. Using this profile, CHROMACAL can: evalute the linearity of your imaging system; and determine the color corrections needed to transform your raw specimen images into colorcalibrated images. Below is a calibration slide image after the color matrix was expanded to fill the Calibration Profile window. Note that the diagnostic results are not sufficient to move forward to color calibrating specimen images NOTE After a calibrated image is saved, you cannot modify the contrast or the Reference Standard Label within the CHROMACAL Image software. In this situation, the View ChromaCal Images function is view-only. You can zoom in/out and around the image to review regions of interest. Diagnostic Measurements of the Calibration Slide Image (Note: the diagnostic measurements are not sufficient to continue with color calibration) Each of the diagnostic measurements (Matrix Alignment, Linearity and Exposure) has a value defining its result. You want to achieve the following diagnostic results before proceeding to capture your specimen images and before proceeding to color calibrate your specimen images: Matrix Alignment: Desired result = "Good" Linearity: Desired result = "Good" or "Within Tolerance" Exposure: Desired result = "Good" When the above results are achieved, you can proceed with color-calibrating your specimen images. If the diagnostic results are otherwise, click the underlined hyperlink provided below each diagnostic category to view guidance on how to proceed (see below). CHROMACAL User Guide (v 1.1) 36

Matrix Alignment The Matrix Alignment result has one of two possibilities: Result Hyperlinked Guidance on Matrix Alignment Result "Good" With this result, no further adjustment s are need to the matrix alignment. "Poor" If you colorcalibrate using a poorly aligned matrix, your calibration results will likely not be optimal. A blue rectangle must be completely contained within each color circle. The exact location within each circle is not critical. If any adjustments are needed, re-align the squares. When you hover the mouse over each square, adjustments are shown. The adjustments vary depending on the square location. The Matrix Alignment must be "Good" before the results of Linearity and Exposure can be relied upon. CHROMACAL User Guide (v 1.1) 37

Linearity Achieve a Matrix Alignment = "Good" before relying on the Linearity result. The Linearity result has one of three possibilities: Result Hyperlinked Guidance on Linearity Result "Good" With this result, you can proceed with colorcalibrating your specimen images. "Within Tolerance" Although your imaging system is not capturing linear images, it is within an acceptable tolerance. "Poor" You cannot colorcalibrate images if your linearity result is poor. Click "Show Linearity Plot" to see a graphical representation of the result. CHROMACAL User Guide (v 1.1) 38

Exposure Achieve a Matrix Alignment = "Good" before relying on the Exposure result. The Exposure result has one of three possibilities: Result Hyperlinked Guidance on Exposure Result "Good" With this result, you can proceed with colorcalibrating your specimen images. "TBD" The exposure cannot be assessed. Be sure matrix is aligned properly. CHROMACAL User Guide (v 1.1) 39

Result Hyperlinked Guidance on Exposure Result "Over Exposed" If you continue with this result, the color calibration will likely not be optimal and the calibrated images will likely be darker than your original images. CHROMACAL User Guide (v 1.1) 40

Tools for Calibrated Specimen Images Whether you are calibrating a single image at a time, or in the Batch Processing function, there are tools available for viewing the image and adjusting specific parameters. NOTE After a calibrated image is saved, you cannot modify the contrast or the Reference Standard Label within the CHROMACAL Image software. In this situation, the View ChromaCal Images function is view-only. You can zoom in/out and around the image to review regions of interest. (Above: Single Image Window) (Above: Batch Processing Window) CHROMACAL User Guide (v 1.1) 41

Viewing Tools To zoom in, zoom out or move around images, use the following Viewing Tools. Magnifier Tool (default tool). Use to magnify the image(s) (or <shift>click to shrink). The cursor will appear as a "+" symbol (or "-" symbol if in shrink mode) as you move the cursor over either image. Hand Tool. Click to activate, and the cursor will change to a "hand" symbol when you drag it over either image. Click and hold the mouse button and to move the entire image around the display window. (Note: images cannot be moved with the Hand tool unless they have first been zoomed-in.) Reset. Return the images to their original size To expand the images within a region of interest: With the Magnifier Tool selected, press and hold the mouse button starting at the top left-hand corner of interest and drag the selection rectangle to the lower right-hand corner of interest. NOTE You can make viewing adjustments to either the calibrated or uncalibrated image. The same adjustment will automatically be replicated on the other image. Just try it! CHROMACAL User Guide (v 1.1) 42

Contrast Slider CHROMACAL Image software calibrates the specimen image at a baseline contrast level. In practice, contrast can be an individual preference based on the specimen under review. CHROMACAL Image software incorporates flexibility for a controlled range of contrast adjustment. Moving slider to the left Moving slider to the right Reduces the contrast level on the calibrated image (depicted as a negative number) Increases the contrast level on the calibrated images (depicted as a positive number) Reset. Resets the calibrated image to the default contrast level of 0 Apply to List. Only available in the Batch Processing function, this feature will apply the contrast level of the current image to ALL images listed in the batch. NOTES The contrast slider only impacts the calibrated image. There is no contrast change made to the original (uncalibrated) image. As the contrast slider is moved, the CHROMACAL contrast feature maintains the linear relationship of the color components (red, green, blue). The default contrast level within the software is set at zero, which will result in the highest level of color consistency. CHROMACAL Image software does not employ a gamma-based contrast adjustment since such a method alters the relationship between RGB in a non-linear manner. Reference Standard Label By default, CHROMACAL adds a Reference Standard Label to each calibrated image. This provides a visual confirmation that the image has color integrity that will remain visible when the image is shared between colleagues, facilities, and in print. The label can provide a level of confidence that the CHROMACAL color standard has been used. CHROMACAL User Guide (v 1.1) 43

[default] Reference Standard Label will be inserted on the calibrated image Reference Standard Label will not be inserted on the calibrated image Location. Specifies the location of the Reference Standard Label on the calibrated image Color. Specifies the text color for the Reference Standard Label on the calibrated image BEST PRACTICES We strongly recommend the use of the Reference Standard Label. Its presence conveys to all parties that your specimen images possess color integrity from utilizing the CHROMACAL Color Calibration System. NOTES The Reference Standard Label is inserted only on CHROMACAL-calibrated images. The original specimen image files are ALWAYS untouched and remain preserved in their original form. View CHROMACAL Images 1. To view your CHROMACAL-calibrated images, access this function and click Select ChromaCal Images Folder to identify the folder that contains the image(s). 2. Upon selecting the folder, a listing of all calibrated images in the folder displays on the left side of the window. CHROMACAL User Guide (v 1.1) 44

Contents of metadata stored in the calibrated image file See Viewing Tools for additional details on zooming, move around the image, and highlighting a region of interest. NOTES Listed below the calibrated image are the contents of the metadata that were added by CHROMACAL. This information provides a full audit trail so that changes made to the original specimen image are clearly identifiable. This function is a "view-only" window, so no adjustments can be made to contrast or the Reference Standard Label. CHROMACAL User Guide (v 1.1) 45

Send Your Feedback At any time, you can provide feedback to Datacolor regarding the CHROMACAL family of products. Feedback can include general questions, product suggestions and troubleshooting information. Click Send Your Feedback and the email client on your workstation will be launched with the "to" address as info.chromacal@datacolor.com automatically completed. We appreciate your feedback and respond to it. Just try it! CHROMACAL User Guide (v 1.1) 46

Menu Bar Options File Menu This menu provides an alternative method for opening the calibration slide image. It also offers other file management options typically accessed only when troubleshooting technical issues with CHROMACAL customer support staff. Open Calibration Slide Image This option opens a dialogue window and prompts you to select the calibration slide image associated with the specimen images you want to color-calibrate. See Open Calibration Image for further instructions. Open CHROMACAL Log Folder This is a primarily a troubleshooting tool for use by Datacolor. It should only be accessed at the request of CHROMACAL customer support. Open CHROMACAL Application Folder This is a primarily a troubleshooting tool for use by Datacolor. It should only be accessed at the request of CHROMACAL customer support. Load Measurement File from Disk This feature will add a measurement file associated with a specific CHROMACAL Calibration Slide to the local measurement file database. This option is needed only if you do not have internet access when you are using CHROMACAL Image. Only use this option at the direction of CHROMACAL customer support. You will need to have access to another workstation with internet access and email capability, so that you can receive the measurement file from CHROMACAL customer support. 1. Click Load Measurement File from Disk. The prompt confirms the action: CHROMACAL User Guide (v 1.1) 47

2. Click Yes. Browse to the folder containing the measurement data for the slide: Quit Edit Menu 3. Highlight the file and click Open. The program will retrieve the measurement file and store it in the local Measurement File database. The option will close the CHROMACAL Image program. The Edit Menu contains standard edit functions, as well as the ability to define certain user and software preferences. The edit functions will only be active when those capabilities are applicable within the program. Preferences To access preferences: 1. Click Preferences. The information below displays: CHROMACAL User Guide (v 1.1) 48

The program identifies the most recent measurement data accessed. This is for informational purposes only. BEST PRACTICES The program by default will automatically check for software updates on launch. THIS IS THE RECOMMENDED SETTING. We endeavor to keep updating the software with performance and efficiency improvements. Keeping this option activated means you will receive the latest update of the program. If there is no checkmark within the box, you are NOT receiving the most recent updates when you launch CHROMACAL Image. View Menu The View Menu features the ability to view calibrated specimen images. It also provides an option to view the statistics regarding images that have been color-calibrated on your workstation. ChromaCal Images To view your CHROMACAL-calibrated images, access this function. Only calibrated images are available under this function, and you cannot modify the contrast nor the Reference Standard Label in this function. See View ChromaCal Images for further details. Usage Stats For informational purposes only. This option will display the number of specimen images calibrated in the current CHROMACAL Image session, as well as the total number of images calibrated since the software was activated on that workstation. CHROMACAL User Guide (v 1.1) 49

Tools Menu You can start the batch process feature from this menu. You can also view the log of Measurement Files stored in your local database, and show the linearity plot of your calibration slide image. Batch Process This option opens the Batch Process window. It opens the same window accessed when you click the Batch Multiple Images button on the Main window. See Batch Process for detailed instructions on this feature. Measurement File Database This option is used to view a log of the Measurement Files currently available in your local CHROMACAL database. You can also use this feature to delete Measurement Files. Typically, you would not delete a Measurement File unless: you are directed to do so by CHROMACAL customer support you accidently assigned the wrong serial number to a calibration slide image Show Linearity Plot This option opens a window to display the linearity result of your imaging system based on an assessment of the calibration slide image. CHROMACAL User Guide (v 1.1) 50

Monitor Menu Calibrate This menu offers a seamless integration to the CHROMACAL Monitor Calibration software, both to calibrate (or re-calibrate) your monitor, or to access tools for analyzing the quality of your monitor. To calibrate (or re-calibrate) your monitor: 1. Click Calibrate. This will initiate the launch of the CHROMACAL Monitor Calibration software. The following two screens will display in succession. Analyze Quality Note: You will need the CHROMACAL Monitor Calibration sensor to calibrate your monitor. If the sensor is not available at your workstation, retrieve it from the original System case (or box if purchased separately). 2. Follow the on-screen instructions. See also Quick Start to Monitor Calibration to calibrate your monitor. To analyze the quality of your monitor: 1. Click Analyze Quality. This will initiate the launch of the CHROMACAL Monitor Calibration software and the following screen will appear. CHROMACAL User Guide (v 1.1) 51

If the CHROMACAL sensor is not available at your workstation, retrieve the sensor from the original system case (or box if purchased separately). You cannot analyze the quality of your monitor without a CHROMACAL Monitor Calibration sensor. Click OK to continue if you have the sensor available, or click Cancel to abort the analysis. 2. After clicking OK, the following screen displays. Follow the on-screen instructions. You can also obtain help information by clicking the Help button on the screen above. CHROMACAL User Guide (v 1.1) 52

Help Menu This menu offers access to various resources to support your use of the CHROMACAL Image software as well as to provide your feedback to Datacolor. About CHROMACAL. Displays the software version currently installed, as well as the license code of the CHROMACAL Image software activated on your workstation, and a ChromaCal-assigned computer ID that may be needed for Customer Support. To close this window, click anywhere on the window. Software Update Check. This feature will check if there is a newer version of the program available for download. It is recommended that you always download the latest version of the software. If you do not have the latest version, you will be notified and prompted to download it. BEST PRACTICES If under Edit/Preferences you have enables "Automatically check for software updates on launch", you will likely have the latest version running on your workstation. Read Me. Opens a window to show important, often late-breaking news about the software. User Guide. Opens a browser window and launches a pdf version of the CHROMACAL User Guide. This guide is located on the CHROMACAL website to ensure that you will be viewing the latest version of the User Guide. CHROMACAL User Guide (v 1.1) 53

Version Notes and Instructions. Displays a summary of recent improvements to the version of the CHROMACAL software installed on your workstation. If there are also important new instructions, they will be included. To close the window, click "X" in the upper right-hand portion of the window. End-User License Agreement. Opens a window and displays a PDF version of the End- User License Agreement that was accepted at time of software installation on your workstation. This is provided for review / reference purposes only. Website Resources. Opens a browser window that displays the Resources page of the CHROMACAL website. This page offers an array of microscopy and CHROMACAL information including: o o o o o o o o o A guide on good microscopy practices Video library (instructional and informational videos) Gallery of specimen images (before and after calibration) News / articles on CHROMACAL Upcoming events where CHROMACAL will be discussed and on display CHROMACAL product brochures Screen shots for CHROMACAL Image Calibration software Screen shots for CHROMACAL Monitor Calibration software Third Party Resources on microscopy and color integrity Registration. Displays the current software registration information on file associated with your workstation installation. This information can be updated as needed within this screen. User Experience Feedback. Launches a user survey screen in which Datacolor requests your feedback on various questions (related to microscopy and/or your experience using the CHROMACAL software). There are multiple, brief surveys and we update the questions periodically. It is possible that a given survey may be repeated, but this is done intentionally as your current responses may provide us with valuable information on trends We appreciate your feedback. Your input drives our ongoing development efforts! Thanks in advance for your contributions. Follow the prompts to complete the survey. BEST PRACTICES If you need access to any CHROMACAL on-line documentation (e.g. user guides, website resources, videos, etc.) but do not currently have internet access at your workstation, just use your mobile device. Our website (www.chromacal.com) is mobile device friendly. So, don't let a lack of internet access leave you "helpless." Access CHROMACAL support documentation on your mobile device! CHROMACAL User Guide (v 1.1) 54