Index Sorting Resources

Index Sorting

Index sorting allows researchers to review the complete phenotype of every single event sorted into a plate and associate that event with all sorted and non-sorted events.

FCS Express allows for the analysis and reporting of index sorting data using Heat Maps along with the standard plot offerings.

*The High Content add-on must be enabled for your license to use heat maps for index sorting data analysis. If you have an active FCS Express 7 subscription and High Content is not enabled, please email us at support@denovosoftware.com.*

FCS Express currently supports index sorting data files from the following instruments (please click links for full support documentation):

Combining Heat Maps and Color Dot Plots to Analyze Index Sort Data

For index sort data, Heat maps will display the number of events (usually one) and parameter-based phenotypic data for all sorted events.

Wells may be gated individually, or as groups, with the well gates backgated onto other plots to view the individually sorted cells in the context of the entire acquired/sorted samples.

All heat map functionality is available for use with index sorting data. FCS Express offers a number of features, including Emphasized Well Gates, to properly analyze Index Sort Data.

Standard Well Gate versus Emphasized Well Gate on Influx Index Sorting Data

Customizing Heat Maps for Index Sort Data

Heat Map Overlays allow a user to view multiple statistics and parameters on a single heat map by adding each statistic as a new overlay.

Additionally, users may add a radius statistic, which changes the size of the well to reflect the selected parameter.

The image below on the left shows the GFP Mean in the lower half and the PE-Cy7 Mean in the upper half. The image below on the right shows GFP Mean as the statistic being used to color each well in the heat map, while the radius of the well is based on the PE-Cy7 Mean.

Instrumentation Examples

Influx Index Sorting

Influx Index Sorting Data Files contain fluorescence information on both the sorted and unsorted events.

A default heat map can be created to display statistics related to a single parameter. A custom heat map can be created to display multiple parameters related to the index sort data.

Well gates can be created and applied to view index sorting data on a color dot plot. Individual well gates can be created to isolate a single sorted event. In addition, a well gate may be created on a group of wells to isolate multiple sorted events.

Heat Map with Radius Parameter

Index Sorted and Non Sorted Events	All Index Sorted Events	Single Index Sorted Event

Diva Index Sorting

Diva Index Sorting Data Files contain fluorescence information only on the indexed sort events.

A "master" .fcs file contains information on all of the events collected during a sort.

These two files may be used individually or in conjunction via plot overlays to help visualize index sorting data acquired with Diva.

A default heat map can be created to display statistics related to a single parameter. A custom heat map can be created to display multiple parameters related to the index sort data.

Well gates can be created and applied to view index sorting data on a color dot plot.

Individual well gates can be created to isolate a single sorted event. In addition, a well gate may be created on a group of wells to isolate multiple sorted events.

Heat Map with Radius Parameter

Index Sorted Data File	"Master" Data File	Index Sorted and "Master" File Overlay

Summit Software Sorting Data

Summit Software Sorting Data Files contain fluorescence information on both the sorted and unsorted events.

A default heat map can be created to display statistics related to a single parameter. A custom heat map can be created to display multiple parameters related to the index sort data.

Well gates can be created and applied to view index sorting data on a color dot plot.

Individual well gates can be created to isolate a single sorted event. In addition, a well gate may be created on a group of wells to isolate multiple sorted events.

Heat Map with Overlay

Index Sorted and Non Sorted Events	All Index Sorted Events	Single Index Sorted Event

Chorus Index Sorting

Chorus Index Sorting Data Files only allow for analysis of indexed sorted events.

A default heat map can be created to display statistics related to a single parameter. A custom heat map can be created to display multiple parameters related to the index sort data.

Well gates can be created and applied to view index sorting data on a color dot plot.

Individual well gates can be created to isolate a single sorted event. In addition, a well gate may be created on a group of wells to isolate multiple sorted events.

Heat Map with Overlay

Index Sorted Events		Index Sorted Events with Well Gates

Index Sorting

Instrumentation Examples

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