FlowJo Enterprise brings significantly enhanced QC to data transfer, assessment of sample quality, and data analysis.
1. Data Transfer
Two different file fidelity checks and re-run and checked against a manifest when data is uploaded to the FlowJo Server to ensure faithful transfer. For more information, please see the Data Management page.
2. Sample Quality
The quality of the sample acquisition as determined by data available in the FCS file can be determined and automated during pre-processing of data files on the server. The sample is examined in its entirety for event rate consistency and on a parameter-by-parameter basis to examine the slope of parameters v. time. Importantly, spreadsheet reports can be generated to examine sample quality over time either automatically, or can be invoked on a particular data set of interest.
Figure 1. Example visualization of parameter medians v. time.
3. Data Analysis
Flow cytometric analysis depends largely on gating (subsetting of cell populations); regardless of whether gates are created by algorithms or human experts their quality and consistency are difficult to assess.
Figure 2. Representative SAMSpectral, K-Means, and Manual Gating/Clustering results.
The FlowDx platform provides the means through a simple GUI interface for a user to select any number of populations, any number of gating results and calculate one or more comparison statistics to examine the accuracy, consistency, and expertise assumption of flow cytometry analysis.
High-throughput facilities, particularly clinical operations and CROs, must employ either multiple technicians or use algorithms to process large numbers of data files daily. In order to examine and quantify technician consistency and algorithm accuracy, and funded as part of a SBIR research grant, we partnered with an immunology laboratory at Harvard to develop and demonstrate a suite of statistical methods to evaluate the quality of the gating. This development partnership allowed us to develop the FlowDx platform, which provides a simple GUI interface for a user to select any number of populations and any number of gating results and calculate one or more statistics on each. The embedded statistics are F-measure, the Davies-Bouldin score, and the Match Ratio, another product of this work. One of the advantages to this platform is that the gate generators can be a heterogeneous mixture of humans and algorithms and that results themselves can be consequently gated to analyze the analyzers.
To learn more about FlowDx, please contact us and visit the project page here.