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
T
R
TRUTH
T
H
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 :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
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