Purpose of the Procedure: This procedure describes the steps involved in upgrading the graphical user interface (GUI) and controller software of an IncuCyte. The goal of the procedure is to update the IncuCyte GUI to 2011A Revision 2 and the IncuCyte controller to 2011A Revision 3. This procedure can also be used to upgrade an IncuCyte currently running 2011A to the latest version. Required Upgrade Packages: IncuCyte 2011A Rev2 GUI Install CD and access to the Internet. If a CD is not available, one of two styles of GUI installers can be downloaded. See the table in the Procedure Overview section for the specific upgrade files. Procedure Overview: The software update of an IncuCyte system to a new base version (eg., 2008A, 2008B, 2009A, etc) requires that you update the GUI at the same time you update the controller software. The steps you need to take to upgrade the software on the IncuCyte will depend on the version that is currently running on the system. The upgrades can be a multi-step process that involves both GUI updates as well as IncuCyte controller (device) updates. The table below is used to walk you through the update steps. Begin by finding the current version of software that is running on the system. Find this version in the left column of the table, then follow the steps indicated in the right column. The guiding principle of upgrades is this: first upgrade to the base version of the release, then follow this with upgrades to any revisions within the base version that have been deployed. The base version is designated by a year and a letter; examples include 2008A and 2009B. A revision is designated as 2009A Rev2, for example. The specific upgrade flow when upgrading to a new base version goes as follows: 1. If an upgrade is available, install the new GUI, but do not uninstall the GUI currently used. 2. If the GUI upgrade is successful, proceed to upgrade the controller software using the original (currently used) version of the GUI (not the newer GUI installed in the previous step). More than one controller update might be required (see the table). 3. Verify the previous step by connecting to the device using the new GUI. Uninstall the older GUI if desired. 4. Optional: Update the vessels database to obtain 96-well ScratchWound and/or 96-well Angiogenesis support for newly licensed users. If you were already running these assays prior to the upgrade, then this step is not required unless you are interested in obtaining the very latest IncuCyte-supported vessels. Page 1
The Upgrade Steps Table below shows the steps you need to take to upgrade the software on the IncuCyte, which depend on the version that is currently running on the system. Find your current version in the left column, then follow the upgrade steps in the right column. Current software version 2011A 2010A, 2009B, 2009A or 2008B (any version) 2008A (any version) Steps for software upgrade Step 1) GUI update: section G2 Step 2) Controller update: section C4 Step 3) Optional vessels database update: section V1 Step 1) GUI update: section G1 Step 2) Controller update: section C2 Step 3) Controller update: section C3 Step 4) Controller update: section C4 Step 5) Optional vessels database update: section V1 Step 1) GUI update: section G1 Step 2) Controller update: section C1 Step 3) Controller update: section C2 Step 4) Controller update: section C3 Step 5) Controller update: section C4 Step 6) Optional vessels database update: section V1 Upgrade Steps Table - If the device is currently running an IncuCyte software version older than 2008A (e.g., R3 or older) or you are unsure of the current software version, then contact Essen BioScience ussupport@essenbio.com. - For a list of features included in each release please refer to the following Software Release Notes: ftp://50.201.185.115/incucyte/incucyte%20software%20release%20notes.pdf NOTES The software upgrade procedures update the IncuCyte software, but do not automatically alter the database of vessels, trays and scan patterns. A unique set of database update files exists for each type of IncuCyte hardware (HD vs. FLR), and it s critical to use the corresponding set of update files. Brand new users of the 96-well ScratchWound and/or 96-well Angiogenesis assays require a vessels database update. As such, you must contact support (ussupport@essenbio.com) to request the appropriate vessel database update and follow the Vessels Database Update procedure (section V1). Page 2
IncuCyte FLR UPGRADES: VERY IMPORTANT o If upgrading the controller to 2011A from 2009A or older, all IncuCyte FLR devices must be recalibrated before the Fast Fluorescence feature can be used! Run the Fluorescence Calibration procedure on the device. Page 3
Procedure Detail: Section G1: Install the 2011A Rev2 IncuCyte User Interface (GUI) Before we install the 2011A Rev2 GUI (2011A for short), here are some important notes: - A 2011A controller will reject a connection from a non-2011a GUI - A 2011A GUI will not connect to a non-2011a controller - A pre-2011a GUI will not connect to a 2011A controller. - The IncuCyte 2011A GUI should be installed side-by-side with any older versions of the IncuCyte GUI. You do not have to uninstall the previous GUI version, and you should not if there are older controllers at the facility. - It is critical to install the 2011A GUI before doing the 2011A controller upgrade (Sections C1, C2, C3, and C4). Installation Instructions: The 2011A IncuCyte GUI is available either on an installation CD or via these Web links: ftp://50.201.185.115/incucyte/2011a/rev2/incucyte2011arev2guisetup.zip [253 Mb] Large, standalone download that does not require Internet connectivity. The Microsoft.NET 3.5 installer is bundled with this installer. The installation of.net v3.5 can take up to 5 minutes. ftp://50.201.185.115/incucyte/2011a/rev2/incucyte2011arev2guiuseswebsetup.zip [47 Mb] Small download for installation on PC s with Internet connectivity. Internet connectivity is required to obtain Microsoft.NET 3.5 via download. Not counting download time, the installation of.net v3.5 alone can take up to 5 minutes. 1. DO NOT UNINSTALL OLDER VERSIONS OF THE INCUCYTE GUI AT THIS POINT! 2. The installation of the 2011A IncuCyte GUI must be done with a Windows account with Administrator privileges. The installation will fail otherwise. 3. The 2011A IncuCyte GUI builds upon new Microsoft.NET technology found in.net Framework 3.5. As such,.net 3.5 is installed if necessary. The installation of.net 3.5 alone can take up to 5 minutes. 4. Double-click the installer EXE of choice (standalone or Web-based) to launch. Depending on the Microsoft components already installed on the PC, the installation could take anywhere from under a minute to up to 5 minutes. Please be patient and allow the Microsoft components to install. 5. Proceed with the next step of the upgrade found in the Upgrade Steps Table. Page 4
Section G2: Upgrade the 2011A IncuCyte Graphical User Interface (GUI) to 2011A Rev2 The instructions in this section apply to those who have already installed a version of the 2011A IncuCyte GUI and are upgrading the GUI to the latest revision (Rev2). If you have not installed a version of the 2011A IncuCyte GUI, please refer instead to Section G1. If you are already running the 2011A Rev2 GUI, then you can skip this step and proceed instead with the next step of the upgrade found in the Upgrade Steps Table. Installation Instructions: The 2011A Rev2 IncuCyte GUI update is available via this Web link: ftp://50.201.185.115/incucyte/2011a/rev2/incucyte2011arev2guilitesetup.zip [29 Mb] Small download for installation on PC s with a version of the IncuCyte 2011A GUI already installed. 1. The installation of the 2011A Rev2 IncuCyte GUI must be done with a Windows account with Administrator privileges. The installation will fail otherwise. 2. If your current 2011A IncuCyte GUI is running, close it before installing the Rev2 update. You must also close any Analysis Job Processors that are analyzing IncuCyte data as well as the Plate Map editor. 3. Extract the installer file from the ZIP archive above and double-click the installer EXE to launch. Depending on the Microsoft components already installed on the PC, the installation could take anywhere from under a minute to as much as 5 minutes. Please be patient and allow Microsoft components to install. 4. The shortcut to the GUI will still be named IncuCyte 2011A and is placed on the Desktop 5. Proceed with the next step of the upgrade found in the Upgrade Steps Table. Page 5
Section C1: 2008A pre-installation update These controller update instructions are for IncuCyte devices that are currently running 2008A software. If you are unsure of the software version you are running, contact Essen BioScience ussupport@essenbio.com. Please make sure that (A) the device is not busy with scanning, calibrating, etc. and (B) other than the GUI used for upgrading, there are no other GUIs connected to the IncuCyte. Upgrade Instructions: 1. Copy ftp://50.201.185.115/incucyte/2008b/2008ato2008bcontrollerupdateprereq.zip to your PC Desktop. 2. In this step, we extract the files from the archive (2008Ato2008BControllerUpdatePrereq.zip). Right-click on 2008Ato2008BControllerUpdatePrereq.zip and select the Extract all option. This will start a file extraction wizard. If you accept all of the defaults in the wizard, then the files will be extracted to a new folder on your desktop named 2008Ato2008BControllerUpdatePrereq. 3. Log onto the older IncuCyte GUI using an account with an Administrator permission level. You will not be able to connect with the 2011A GUI. 4. Verify that the device is Idle and there are no imminent scans scheduled. We do not recommend applying an upgrade when the Device Status is displayed with red-faced type! Also verify that other users are not connected to the device from other computers. Administrative activities like archives, etc. will be disrupted by the upgrade. 5. In the IncuCyte GUI, enter the Administer IncuCyte view (under the Task List in the upper left), and select the Update tab. 6. Press the Update IncuCyte button and browse to the folder (e.g., Desktop\2008Ato2008BControllerUpdatePrereq) containing the upgrade files. After selecting the 2008Ato2008BControllerUpdatePrereq folder, press the OK button to upload the update files to the device. Confirm that you want to proceed with the upgrade. 7. The upgrade procedure takes about 2 minutes. You will need to disconnect from the device during the upgrade. Either close the GUI or just close the connection. 8. After two minutes, reconnect to the device using the current (older) GUI. You will not be able to connect with the 2011A GUI at this point. After verifying that you can connect, proceed with the next step of the upgrade found in the Upgrade Steps Table. Page 6
Section C2:.NET 2.0 SP2 pre-installation update These controller update instructions are for IncuCyte devices that are currently running 2010A, 2009B, 2009A, 2008B, or 2008A software. If you are unsure of the software version you are running, contact Essen BioScience ussupport@essenbio.com. Please make sure that (A) the device is not busy with scanning, calibrating, etc. and (B) other than the GUI used for upgrading, there are no other GUIs connected to the IncuCyte. Upgrade Instructions: 1. Copy ftp://50.201.185.115/incucyte/2009b/dotnet2sp2andupdate.zip to your PC Desktop. 2. In this step, we extract the files from the archive (DotNet2SP2AndUpdate.zip). Right-click on DotNet2SP2AndUpdate.zip and select the Extract all option. This will start a file extraction wizard. If you accept all of the defaults in the wizard, then the files will be extracted to a new folder on your desktop named DotNet2SP2AndUpdate. 3. Log onto the older IncuCyte GUI using an account with an Administrator permission level. You will not be able to connect with the 2011A GUI. 4. Verify that the device is Idle and there are no imminent scans scheduled. We do not recommend applying an upgrade when the Device Status is displayed with red-faced type! Also verify that other users are not connected to the device from other computers. Administrative activities like archiving, etc. will be disrupted by the upgrade. 5. In the IncuCyte GUI, enter the Administer IncuCyte view (under the Task List in the upper left), and select the Update tab. 6. Press the Update IncuCyte button and browse to the folder (e.g., Desktop\ DotNet2SP2AndUpdate) containing the upgrade files. After selecting the DotNet2SP2AndUpdate folder, press the OK button to upload the update files to the device. Confirm that you want to proceed with the upgrade. 7. The upgrade procedure takes about 7 minutes and involves an IncuCyte controller reboot. You will need to disconnect from the device during the upgrade. Either close the GUI or just close the connection. 9. After seven minutes, reconnect to the device using the current (older) GUI. You will not be able to connect with the 2011A GUI at this point. Then proceed with the next step of the upgrade found in the Upgrade Steps Table. Page 7
Section C3: 2011A controller update These controller update instructions are for IncuCyte devices that are currently running 2010A, 2009B, 2009A, 2008B, or 2008A software. If you are unsure of the software version you are running, contact Essen BioScience ussupport@essenbio.com. IMPORTANT: Although the current IncuCyte GUI is used in this step, we strongly recommend that you install the new 2011A GUI before performing this step of the controller update. The IncuCyte controller must be updated using the current (older) GUI, not the new GUI. After the controller is updated, you can only connect to it with the new 2011A GUI. If the new 2011A GUI installation fails, then you do not want to update the controller. 2011A GUI installation instructions are found in Section G1. Please make sure that (A) the device is not busy with scanning, calibrating, etc. and (B) other than the GUI used for upgrading, there are no other GUIs connected to the IncuCyte. Upgrade Instructions: 1. ftp://50.201.185.115/incucyte/2011a/base/2008aorgreaterto2011acontrollerupdate.zip should be copied to your PC Desktop. 2. In this step, we extract the files from the archive (2008AOrGreaterTo2011AControllerUpdate.zip). Rightclick on 2008AOrGreaterTo2011AControllerUpdate.zip and select the Extract all option. This will start a file extraction wizard. If you accept all of the defaults in the wizard, then the files will be extracted to a new folder on your desktop named 2008AOrGreaterTo2011AControllerUpdate. 3. Log onto the older IncuCyte GUI using an account with an Administrator permission level. You will not be able to connect with the new 2011A IncuCyte GUI. 4. Verify that the device is Idle and there are no imminent scans scheduled. We do not recommend applying an upgrade when the Device Status is displayed with red-faced type! Also verify that other users are not connected to the device from other computers. Administrative activities like archives, etc. will be disrupted by the upgrade. 5. In the IncuCyte GUI, enter the Administer IncuCyte view (under the Task List in the upper left), and select the Update tab. 6. Press the Update IncuCyte button and browse to the folder (e.g., Desktop\2008AOrGreaterTo2011AControllerUpdate) containing the upgrade files. After selecting the 2008AOrGreaterTo2011AControllerUpdate folder, press the OK button to upload the update files to the device. Confirm that you want to proceed with the upgrade. 7. The upgrade procedure takes about 5 minutes. Once the upgrade starts, exit the GUI. In the next step you will reconnect using the new 2011A IncuCyte GUI. If you haven t installed this, you must see the instructions above. 8. After five minutes, reconnect to the device using the new IncuCyte 2011A GUI. You will not be able to connect with the older GUI at this point. Once connected, verify the upgrade using the Help -> About menu item. The Device information box in the center of the window should contain the following entries (among many others): IncuCyte Software Build: 2011A Page 8
Section C4: 2011A Revision 3 controller update These controller revision update instructions are for IncuCyte devices that are currently running the 2011A base controller software. If your IncuCyte is already running 2011A Rev3, then you do not need to apply this update! If you are unsure of the software version you are running, contact Essen BioScience ussupport@essenbio.com. The IncuCyte controller must be updated using the 2011A GUI or any revision of the 2011A GUI. 2011A GUI installation instructions are found in Section G1. Please make sure that (A) the device is not busy with scanning, calibrating, etc. and (B) other than the GUI used for upgrading, there are no other GUIs connected to the IncuCyte. Upgrade Instructions: 1. Copy ftp://50.201.185.115/incucyte/2011a/rev3/2011arev3controllerupdate.zip to your PC Desktop. 2. In this step, we extract the files from the archive (2011ARev3ControllerUpdate.zip). Right-click on 2011ARev3ControllerUpdate.zip and select the Extract all option. This will start a file extraction wizard. If you accept all of the defaults in the wizard, then the files will be extracted to a new folder on your desktop named 2011ARev3ControllerUpdate. 3. Log onto the IncuCyte 2011A Rev2 GUI using an account with an Administrator permission level. 4. Verify that the device is Idle and there are no imminent scans scheduled. We do not recommend applying an upgrade when the Device Status is displayed with red-faced type! Also verify that other users are not connected to the device from other computers. Administrative activities like archives, etc. will be disrupted by the upgrade. 5. In the IncuCyte GUI, enter the Administer IncuCyte view (under the Task List in the upper left), and select the Update tab. 6. Press the Update IncuCyte button and browse to the folder (e.g., Desktop\2011ARev3ControllerUpdate) containing the upgrade files. After selecting the 2011ARev3ControllerUpdate folder, press the OK button to upload the update files to the device. Confirm that you want to proceed with the upgrade. 7. The upgrade procedure takes about 2 minutes. Once the upgrade starts, exit the GUI. In the next step you will reconnect using the IncuCyte 2011A Rev2 GUI. If you haven t installed this, you must see the instructions above. 8. After three minutes, reconnect to the device using the IncuCyte 2011A Rev2 GUI. Once connected, verify the upgrade using the Help -> About menu item. The Device information box in the center of the window should contain the following entry (among many others): IncuCyte Software Build: 2011A Rev3 Page 9
Section V1: Vessels Database Update A vessels update is required for new 2011A users of the 96-well ScratchWound and/or 96-well Angiogenesis assays. If you have already been running either of these assays successfully prior to upgrading your software, then this vessels update is not required. The vessels update is also available to licensed customers who want the very latest set of IncuCyte-supported vessels. If you are unsure about whether or not you need this update, please contract Essen Support. A zip archive obtained from Essen BioScience Support is required to perform a database update. A unique set of database update files exists for each type of IncuCyte hardware (Plus, HD, FLR, EX, etc), and it s critical to use the correct set of update files. Furthermore, the set of supported vessels at times depends on the version of software running on the IncuCyte. As such, you must contact Essen BioScience Support (ussupport@essenbio.com) to request the appropriate set of vessel database update files. For the purpose of these instructions, we ll assume that the archive obtained from EssenBioScience is named VesselsUpdate.zip. 1. Begin by copying the file VesselsUpdate.zip to your PC Desktop. 2. In this step, we extract the files from the archive (VesselsUpdate.zip). Right-click on VesselsUpdate.zip and select the Extract all option. This will start a file extraction wizard. If you accept all of the defaults in the wizard, then the files will be extracted to a new folder on your desktop named VesselsUpdate. Inside this folder you will find at least 4 database update files: Disks.xml, Trays.xml, ScanPatterns.cfg, and VesselTypes.xml. There might be a 5 th file as well, called UserInfo.dat. 3. Log onto the IncuCyte Graphical User Interface (GUI) software using an account with an Administrator permission level. 4. Verify that there are no scans running currently, and no scans pending to start within the next 5 minutes. We do not recommend applying an update when the Device Status is displayed with red-faced type! 5. In the IncuCyte GUI, enter the Administer IncuCyte view (under the Task List in the upper left), and select the Update tab. 6. Press the Update Database button and browse to the folder (e.g., Desktop\VesselsUpdate) containing the update files. After selecting the VesselsUpdate folder, press the OK button to upload the set of new database files to the device. You will get a confirmation message when complete. This might take up to 30 seconds, so please have patience while you wait for confirmation. There will be no feedback during the update, but a message will appear when complete. If the message contains an error, please contact Essen BioScience Support ( ussupport@essenbio.com ). 7. Navigate to the Schedule Upcoming Scans view in the GUI and press the Reload button in the bottom right corner to refresh your vessel and tray lists. Press OK when asked if you want to continue. 8. Confirm that the update went through by looking for the expected set of new vessels and perhaps trays. 9. Finally, for those customers using either 384-well or 96-well microplates, we recommend that you re-run the 384-well Plate Calibration through the GUI in order to take advantage of the improvements to the 384-well microplate definitions. If you have any questions about how to run the 384-well Plate Calibration, then consult the IncuCyte GUI User Manual and/or contect Essen BioScience Support (ussupport@essenbio.com ). Page 10