A routine is an FCAP-list document which saves a data analysis method that you develop for batch analysis of listmode data acquired on samples which have been prepared with similar protocols and reagent combinations. An FCAP-list routine stores information for the classification of populations, i.e. one gate set for each target population, and it determines how the analysis results are saved and recorded. In order to create a routine you should perform the tasks you want to automate and save the recorded steps into a disk file.Similar to the routines, projects are also data analysis routines. However, in contrast to a routine which is a 'static object' that you cannot change during the execution, projects provide interactive data analysis. The projects are essentially 'stand-alone programs' written in a special programming language called FCAP-BASIC. This language has been developed for the flow cytometry data analysis and processing. The language allows you to code complex data analysis protocols with sophisticated mathematical algorithms and provide them for othe laboratories and users who are interested in applying these methods. FCAP-list provides an interpreted environment to execute these programs as projects.
Projects are complex analysis tools that may contain several routines and commands. Each project may have several input data files, and the results of previous calculations may be used in subsequent ones.
You can find several examples of the projects in the FCAP-BASIC section.
Populations and Gates
The fundamental objects created and manipulated in a routine are population-defining one- and two-dimensional gates that select and process the events of a population. On the screen, you define populations by graphical manipulation of linear or polygon gates in a set of one-dimensional histograms and two-dimensional dot plots.The association between a population and the gate set that defines it is the color of the gates. Gates of the same color define the same population of events. The gates can be placed on histograms using the subset of parameters which best defines the population.
Up to 8 populations can be defined simultaneously. Populations need not be conceived of as mutually exclusive groups.
Smart Gates
![[IMAGE]](2psmart.gif)
One of the most important benefits of using FCAP-list lies in the use of the so-called Smart Gates. In contrast to the traditional "fixed-region" gates the Smart Gates are able to offset their positions and correct day to day or experiment to experiment shifts in populations caused by instrument drift, differences in sample preparation, etc. The main advantage using Smart Gates is the ability given you to perform automatic data analyis routines on a series of slightly different samples. The use of Smart Gates eliminates the need for manual gate readjustments.
Timegating: Kinetic vs. Time
FCAP-list allows you to perform 'kinetic versus time' data analysis on listmode data files. If the data were acquired without recording the time parameter, FCAP-list can generate a 'virtual time' parameter by assuming that the cells were acquired at a costant rate. The timegating routine can handle the events acquired between any time-limits as a distinct population.
Saving Results
FCAP-list allows you to save the analysis results in three formats:
- The Report document saves statistics resulting from the data analysis and information identifying the origin of the reported results in a text file.
- Selected statistical results can be saved in spreadsheet document for further processing the data with any popular spreadsheet applications including Microsoft Excel, Claris Works, etc. With FCAP-list you can convert listmode data and histogram data into ASCII text, and save the converted files in spreadsheet format.
- FCAP-list generates excellent graphics what you can save into PICT files. The Graphic Page Output document, for example, allows you to save histograms and dual parameter distribution plots with the principal analytical results (gate and quadrant statistics) into a PICT file. See FCAP-list Graphics for a few examples.
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