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inverted CommasI can’t recommend FloCyte enough to people. I wish everyone who prepares flow samples – even if they don’t run them – would take FloCyte’s courses

 

 

Data Analysis for Flow Cytometry

Our Data Analysis course will benefit you and your users no matter what software program you choose for analysis.  Our aim is to help you understand how data is generated, presented, and analyzed in the most P1050087accurate, appropriate way.  We can couple this training lecture day with individual or small group software training and would be happy to talk to you about how we can best benefit your flow education!  Join us….

I. In the Beginning…

A.  30,000 foot view of day

B.   History of data in flow

1. Early limitations to generation, storage, analysis and display

2. Standardization

3. Evolution and Expansion of formats

C.   Breaking down the barriers

1. Disk space and the cloud

2. Processing power and the quantum leap

3. Comprehension and acceptance

D.   The fundamental question

1. What are we asking of our data?

2. How do we get it to say what we want it to say?

II. Input

A.  Controls, controls, controls!

1. Compensation controls

2. Gating controls

3. Experimental controls

B.  Instrument setup and characterization

1. The importance of instrument QC

2. Methods of QC

3. Post hoc troubleshooting

C.  Sample quality and relevance

1. A word about sample quality

2. Troubleshooting GIGO

D.  Runtime decisions (Software setup)

1. Parameters to include

2. Pitfalls to avoid

E.  Big Data! (How much do I record?)

1. Counting statistics and resolution

2. Balancing resolution and processing power

III. Output

A. The illusion of image

1. What’s the plot?

2. Which one do I use (why, and why not)?

3. How do I make it pretty (and understandable)?

4. Can you take me higher (dimensionality)?

B. The reality of numbers

1.Lies, damned lies, and….

2. Common numerical extractions and their uses

3. Advanced statistical tests

4. Turning numbers back into graphics

C. The needs of the people and plausible deniability

1. Data analysis for discovery

2. Data analysis for publication

3. Data analysis for presentation

4. When enough is enough

D. Bringing it all together

1.  Making the final report

2.  A brief tour of packages

IV. What lies between

A. The application of compensation

1. Theory of spectral overlap

2. Practice of inverted matrix generation

3. Troubleshooting compensation

B. The definition of gating

1. What is a gate versus a region versus a polygon versus…

2. Hierarchy or Boolean

3. Controls revisited

4. The power of backgating

C. The conceptual paradigms of gating approaches

1. Top down…

2. … or  bottom up?

3. Historical perspectives

D. The return of the big question

1. Does my strategy answer my fundamental experimental question?

V. Curtain call

A. Acquisition SW versus Analysis SW

1.  What’s the difference?

2.  Why don’t I do both?

B. Who are the players?

1.  A non-comprehensive list of flow software

2.  A brief discussion of choice

C. What does the future hold?

1. Hardware enabled, and hardware limited

2. Automation

Questions and answers

Software Training options

This course can be coupled with software training on FlowJo or FCS Express either a second day, or by adding these software labs between lectures

Introduction to the software program

Compensation / Five Color Panel

Data Analysis / Eight Color Panel / Gating, FMO’s etc

Batch Analysis / Templates / Statistics