Flow Cytometry

Data Analysis in a Flash

Joyce Slusser, PhD

KCAS Bioanalytical& Biomarker Services
Shawnee Kansas USA
For Beckman Coulter Life Sciences

FLOW-2139WEB12.16
Background KCAS:
Contract Research Organization
Operate in a GLP environment
Crystal City guidelines dont cover flow cytometry
but are applied to biomarker studies in our facility
Study samples from across the globe
Stability of markers needed for client studies
Logistics, holidays, and inevitable delays
Sample collection and shipping:
Evaluate EDTA, Heparin, ACD and BCT Streck
4hr, 24hr, 48hr, 96hr, 144hr, 192hr, 240hr (10days)
 Sample collection and shipping:
Evaluate EDTA, Heparin, ACD and BCT Streck
4hr, 24hr, 48hr, 96hr, 144hr, 192hr, 240hr (10days)

Sample processing and staining:

Utilized DuraClone IM panels Beckman Coulter
allowed us to eliminate staining inconsistency
Tube Name FITC PE ECD PC5.5 PC7 APC APC-AF700 APC-AF750 PB KrO

TBNK CD16 CD56 CD19 OPEN CD14 CD4 CD8 CD3 OPEN CD45
T subsets CD45RA CD197/CCR7 CD28 CD279/PD1 CD27 CD4 CD8 CD3 CD57 CD45
TCR TCR TCR HLA-DR TCR V1 CD4 CD8 CD3 TCR V2 CD45
B cells IgD CD21 CD19 OPEN CD27 CD24 OPEN CD38 IgM CD45
DCs CD16 LINEAGE OPEN CD1c CD11c Clec9A CD123 OPEN HLA-DR CD45

DuraClone IM panels for Research Use Only, Not for Diagnostic Purposes.
 Goals of todays presentation
Large files easily manipulated
Link gates across study samples
Generate reports while retaining the raw data in the
same file
Higher throughput analysis with statistics reporting
on board
Maximize data output utilizing composites
 Goals of todays presentation
Large files easily manipulated
Link gates across study samples
Generate reports while retaining the raw data in the
same file
Higher throughput analysis with statistics reporting
on board
Maximize data output utilizing composites

Evaluation of sample/biomarker stability:

Launch Kaluza 1.5a software to demonstrate template analysis
Kaluza Analysis Software is for research only and should not be used for diagnostic purposes.
 Summary of Tools Presented
Generate a template
Ease of labeling
Zooming into data
Linking data sets
Linking gates
Using the information tool to create statistics
Creating tables and graphs in Kaluza
Creating reports
Exporting data to excel
Data Summary Examples
COLLECTED IN EDTA

BEST REPORTING:
OVERALL INTEGRITY 48HR
CD25+ 96HR
LYMPH DISTRIBUTION 144HR
COLLECTED IN HEPARIN

BEST REPORTING:
OVERALL INTEGRITY 24HR
CD25+ 192HR
LYMPH DISTRIBUTION 240HR
COLLECTED IN BCT

BEST REPORTING:
OVERALL INTEGRITY 96HR
CD25+ 312HR
LYMPH DISTRIBUTION 360HR
COLLECTED IN ACD

BEST REPORTING:
OVERALL INTEGRITY 96HR
CD25+ 48HR
LYMPH DISTRIBUTION 360HR
 Conclusions
Toolbars in Kaluza are easy to access and use
Kaluza templates allow for reproducible analysis
Kaluza allows manipulation of large data files and
data sets
Merging files
Making composites
Linking gates across data sets
Incorporation of tables into the report format allow
for quick and easy data reprocessing
No need to reprocess and export data
Data review is straightforward
Kaluza Analysis Software is for research only and should not be used for diagnostic purposes.
 Questions?
Presenter: Joyce Slusser
joyce.slusser@kcasbio.com
http://kcasbio.com/