The Art and Science of Flow Cytometry Life in a Parallel Universe John Daley Director Flow Cytometry Hematologic NeoPlasia Dana-Farber Cancer Institute

Definitions Science The classification of knowledge gained by observation and experimentation TRUTH TRTHTRTH ART The seeking for an Ideal truth

What makes a Good Sort Purity Recovery Integrity

What makes a Good Sorter Reliability Reproducibility Responsiveness Stability

The Other Part of the Picture People Operator: Investigator: Supporters: Places: Environment makes it work

Critical Mass Multi application exposure Large user base generates support and application development Equilibrium dynamics coexist between instrument, operators, support, and users Interaction dynamics between Experiment, cells, instrument, researchers, and operators Active Focus on priority at hand: Sort in Progress Not too Big not too small

When things go Right Keep going, don’t look back If it works: Duplicate, duplicate, duplicate Be on the lookout for hidden hisses and the sounds of sparks Stay calm Keep a mindful eye on the relevant features Trust your senses! Be humble

When things go not so Right Stay calm Think what are the components of the issue Try the easiest thing first Remember past experiences Employ back up system if possible Start from beginning Replace the o ring Reboot the computer

The Rituals of Flow Startup Shutdown Keeping control Sort Set up Active Sort Post Sort Sterilization

When the Sort works well Everybody is happy Still need to follow cells down the data path The Proof is in the Pudding Faith in the System, Operator and Protocol Creates confidence for the next experiment Proves Preparation was worth it

How we did it : Open door policy as much as possible Educate ourselves and users about the science and technology Segregate analysis and sorting Maintain as sterile as an environment as possible Tried to keep equipment as up to date as possible Constant form is maintained by constant growth Created a long range Plan Created a concept of supported self service system Always had a back up instrument system in place Always placed experiments on appropriate instruments Learned from our mistakes in trusting others

Time for Expansion 1999: need for high speed and space reorganization Room consolidation : infrastructure Due diligence for instrument best suited for our needs: Critical mass established: need created a confidence that more powerful equipment would be utilized and self supported

Facility Structure Organize User priority Create a common information system: web page/ scheduler Expand tentacles of facility via extended analysis Creating an internal accounting system that could serve as a check in rectifying discrepancies between vendors and administration

Don’t be an Operator Be a Scientist Educate Information resource Never say No (most of the time) Try not to get taken advantage of Mutual Respect is key Above all Maintain a sense of Humor

The Good the Bad and the Ugly When private flow went core When operators were considered paying employees When the bottom line of cost recovery creates an atmosphere of greed Is flow a business or a science?

How to make a sort work It’s in the set up What's needed: operator decides, let user be part of the design, operator maintain transparency Always do a final pre check right before liftoff Transfer a sense of calm to the responsible parties Be part of the sort