HYFLAM MINI-PROJECT REPORT/HANDBOOK OCTOBER 2011 This report/handbook reflects the agreement of the HYFLAM project consortium on the experimental tasks to be included in the project. The report is based on the discussions in the kick-off meeting held at HPA Porton on 14 October 2011. Aims and Objectives The aims and objectives of the project are to identify tasks currently undertaken during experimental procedures with dangerous pathogens in a class III containment cabinet that can be performed by a robotic hand, thus providing an improvement to the efficacy of work undertaken in the cabinet and improve the safety of the operator. The tasks that were discussed include: Opening glass vials containing potentially dangerous pathogens. and posing a significant risk to the operator from injury due to broken glass and sharp edges. Handling a range of tubes and vials. Assisting an operator in cell culture and virus propagation techniques. Preparing diagnostic samples (blood, serum, other body fluids) containing dangerous pathogens for nucleic acid/polymerase chain reaction analysis Handling eggs for injection with dangerous pathogens. Benefits of using a Robot The benefits of using a robot hand were discussed. The main identified benefits are: The robot Hand would help address operator fatigue. Operator fatigue is one of the top reasons for accidents in containment laboratories. Loss of dexterity from working in a glove box cabinet means that even the simplest of tasks, such as opening vials and pipetting, takes twice as long than on an open laboratory bench. Performing dangerous procedures which pose a serious risk of injury to the operator and potential exposure to dangerous pathogens. These include opening sealed glass vials and tasks that include the use of scalpels and needles. Using gauntlets can be difficult as they are normally specified for the largest hands in the lab making them the wrong size for everyone else. The robot hand could remove the need for these entirely. The advantage of a robotic Hand would be that it could perform many different tasks. Most current robotic products are single task/high volume applications. Scenarios A number of possible scenarios were discussed and demonstrated within the laboratory. The agreed scenarios will include: Cell culture and viral propagation: A robotic hand could assist the operator by aspirating fluid from a cell monolayer (requiring care so that the cell monolayer is not disrupted), removing cells from the monolayer and seeding a new cell culture flask, pipetting cells into flasks and cell culture plates (6-, 24and 96-well plates), infecting a cell monolayer with virus and collecting infected cell culture fluid. Loading tubes for centrifugation. This is a procedure that requires precision, as an imbalanced centrifuge can lead to serious accidents. Centrifuge buckets need to be loaded with a precise amount of material and then placed inside a centrifuge bucket without spillage. Buckets than need to be sealed. Opening centrifuge buckets after centrifugation is also a task that needs care and precision so that the material within the centrifuge tube is disturbed as little as possible. Often, after centrifugation, the lids on centrifuge buckets are particularly difficult to open due to the pressures exerted during centrifugation that enforce a seal on the lid. The maximum force required for a task is thus probably opening a centrifuge bucket. Preparing a sample for diagnostic procedures at Biosafety Level 2. Diagnostic samples, including blood, serum, urine, faeces, tissue samples that may contain a dangerous pathogen need to be processed so that they can be handled at BSL2. A robotic hand would allow more samples to be processed to increase throughput and therefore efficiency. Samples are de-activated by extracting the nucleic acids (DNA/RNA) for PCR analysis. Injecting eggs for growth of dangerous pathogens. Some pathogens have very limited growth conditions outside their host and grow best in the laboratory when injected into fertilised hen’s eggs. The procedure is considered to be particularly dangerous as it involves needles to inject the egg with the pathogen and the risk of dropping and breaking the egg during handling is a considerable aerosol hazard. Using a robotic hand would considerably reduce this risk, especially if the whole procedure, from injecting the egg to harvesting the infectious material after incubation, could be hands-free. This procedure was not demonstrated during the kick-off meeting but is considered to be particularly important. Shadow have been provided with various containers/tubes used for the above tasks but will require HPA to provide pipettes and any other equipment needed for evaluation. Other scenarios that were discussed but will not be included in this project for ethical and logistical reasons include using the robotic hand to handle animals housed within containment cabinets. Sterilisation The issue of sterilisation and decontamination of the robotic hand was discussed. The Hand would need to be decontaminated at the end of every experimental procedure. Currently, formaldehyde gas is used at HPA Porton within containment cabinets but this requires high humidity and formaldehyde is potentially corrosive. Sterilisation techniques discussed were: o o o Formaldehdye Hydrogen peroxide Chlorine dioxide Whichever method is used, it must be compatible with all working parts and materials used in the Hand such that the Hand is not compromised or degraded over time. Suppliers of hydrogen peroxide disinfection systems include Steris & Bioquell Some work has been conducted at HPA Porton on the use of low temperature sterilisation methods for spacecraft and spacesuits for the European Space Agency. HPA can request that this information be used to inform this project. The issue of the cleanability of the Hand was raised. It maybe easier to keep the Hand clean if it had a glove cover (this could be disposable). Anything dried onto a hard surface is difficult to sterilise/clean e.g. blood. Removal of overt contamination could be conducted through washing the surface before sterilisation. Another approach maybe to consider a more open frame design for the Hand making it easier to clean but this is outside the scope of this project. Outcomes Demonstration that the use of a robotic hand can improve processes and safety within biological containment o Reduce operator fatigue o Be compatible with current equipment and techniques o Show improvement on current outcomes in terms of: Higher throughput/sample processing Improvement on efficacy and performance Demonstrate ease of use and manageable operator interface. Identification of additional tasks and scenarios for a robotic hand in other areas of biological containment and manipulation Next steps and Tasks Shadow and HPA to agree on scenarios to be included in the project. University of Hamburg to work on a ontology Submission of an abstract for the next EBSA meeting in April.