The Art and Science of Flow Cytometry

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The Art and Science of Flow Cytometry

• Life in a Parallel Universe

John Daley Director Flow Cytometry Hematologic
NeoPlasia Dana-Farber Cancer Institute
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Definitions

• Science
• The classification of knowledge gained by
  observation and experimentation

T R T H
TRUTH
ART The seeking for an Ideal truth
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What makes a Good Sort

• Purity
• Recovery
• Integrity

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What makes a Good Sorter

• Reliability
• Reproducibility
• Responsiveness
• Stability

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The Other Part of the Picture

• People
• Operator
• Investigator
• Supporters
• Places Environment makes it work

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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

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When things go Right

• Keep going, dont 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

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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

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The Rituals of Flow

• Startup
• Shutdown
• Keeping control
• Sort Set up
• Active Sort
• Post Sort
• Sterilization

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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

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How we did it 1996-2007

• 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

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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

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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

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Dont 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

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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?

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How to make a sort work

• Its 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