Installation of a new operating system and reinstallation of EVOware ®

software

After an EVOware® software upgrade, it was discovered that the configure tool was no longer available. The configure tool is used to change fundamental instrument options. It controls the external devices, such as the CP, incubators, plate washer and centrifuge etc. The inability to use this tool meant that the settings on these devices could not be altered. In addition, access to user management functions was also unavailable. Removal of the software upgrade and its reinstallation did not rectify the situation. This procedure resulted in an even more serious issue, which was the inability to start any EVOware® processes. As no other alternative was available, a complete system reinstallation was undertaken on the advice of the TECAN software specialist. This procedure is not trivial and meant installing a new copy of the Microsoft®

Windows® _{XP operating system (OS) on to the PC. In order to preserve the original}

data, the hard drive of the PC was divided, and a new copy of the OS was installed into a separate partition. In addition, to ensure no loss of data from the old system, a number of backup measures were undertaken. The first step was to export the database from the old system using the EVOware®_{export/import tool. This procedure was used to backup}

all processes and labware from the instrument. However, in order to backup specific devices and their drivers, a copy of the whole Tecan folder was saved to an external hard drive. Once the new OS was in place, the next step was to install the new version of EVOware® _{(2.1 SP3). It was at this stage that a number of problems arose. The first}

problem was the inability of the software to communicate with the instrument. This was eventually solved when Com Port 1 was chosen as the instrument’s communication port (Figure 3.2.14). However, another problem arose when testing the devices which were attached to the instrument. Specifically, the centrifuge, the barcode scanner and the incubators could not communicate with the instrument (Figure 3.2.15). Each of the instrument peripherals was plugged into the edgeport device (which connects them to the PC) and after the installation of the edgeport driver the communication problem was partially fixed. The next step was to set proper com settings for each of the peripheral devices (Figure 3.2.16). Once this was completed, the peripheral devices communicated fully with the Tecan instrument. The last two software packages to be installed were the winwasher and the plate reader software. Although the old database was re-imported successfully, software problems still arose with some of the processes.

To overcome this, each process (Figures 3.1.1 and 3.1.2) was rebuilt from within the new version of EVOware®_{. When each process was tested, it ran without producing any}

errors. In addition, all processes could be linked together and run as a single unit.

Figure 3.2.14 The EVOware® _{communication tool was used to check the status of the} robotic instrument. Panel A: After installation of a new OS, the EVOware® _software

could not communicate with the instrument. Panel B: COM port 1 was identified as the proper instrument communication port. The command M1RFV was issued using the send utility and communication between EVOware® _{software and the instrument was}

Figure 3.2.15 There was no communication between the Tecan instrument and its

peripheral devices. None of the peripheral devices (colony picker, centrifuge etc.) were

detected by EVOware®. This problem was partially fixed when the driver for the Edgeport device was installed.

Figure 3.2.16 Final Tecan COM port settings for the peripheral devices. Each

peripheral device has its own com port. The robotic system instrument communicates to the PC via COM port 1 (panel A). The peripheral devices: barcode scanner, centrifuge, colony picker, plate reader, plate washer, incubator and incubatorB communicate on COM ports: 5,8,10,7,6,9 and 4, respectively (panel B).

3.3 Conclusions

Recent advances in screening technology and automation have led to the development of high-throughput systems for use in life sciences. The development of a high-throughput system for the identification of novel affinity reagents from phage libraries has certain advantages. For example, such a system could enable the rapid selection of relevant antibody-producing clones. In addition, electronic-sample tracking and sample handling would allow for the easy identification and backup of superior antibody candidates. Moreover, extremely large numbers of antibody-producing cells could be analysed simultaneously. However high-throughput systems are often complex and, consequently, they are difficult to install, run and maintain. As exemplified in this chapter, installation and testing of a high-throughput antibody screening system was far from trivial.

After assembly and installation of the robotic antibody screening system at DCU, testing revealed a substantial number of hardware and software faults. Hardware problems included the insufficient length of the Tecan robotic arm and colony picker pins, the defective Y-axis motor of the robotic liquid handling arm and the misaligned robotic finger gripper. Although these, and other hardware issues, caused a number of significant difficulties, once they were identified they were readily resolved. In contrast, software problems were extremely difficult to troubleshoot and took far more time and effort to resolve. Major software problems occurred with the StoreX incubator drivers, Tecan EVOware®_{package and the colony picker drivers. In addition, corruption of the}

EVOware®_{database forced a complete PC reinstallation of the Windows}® _{XP OS. Due}

to time constraints, no further work was performed on the robotic antibody screening system. However, all outstanding software and hardware issues were satisfactorily resolved.

Chapter 4

Generation of avian anti-sialic acid