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Flowjo ashland5/11/2023 Summary of Changes Support for Mixed Numeric Data Types This Technical Note provides a summary of changes and guides implementors through some of the new features of the FCS 3.2 specification. It will also be posted at ISAC website upon the publication of this manuscript. The normative version of the FCS 3.2 specification can be found in the Supporting Information SI as well as at. Here, we present FCS 3.2, which is a revision addressing such needs and also incorporating a decade of suggested improvements from the cytometry community at large. The Flow Cytometry Data File Standard was published in 1984 ( 1) to facilitate interoperability of flow cytometry data analysis software with data acquired on different types of flow cytometry instruments. © 2020 International Society for Advancement of Cytometry A reference implementation of the cyclic redundancy check (CRC) calculation is provided in two programming languages since a correct CRC implementation was problematic for many vendors. Types of measurements were formalized, several keywords added, others removed, or deprecated, and various aspects of the specification were clarified. In addition, keywords explicitly specifying dyes, detectors, and analytes were added to avoid having to extract those heuristically and unreliably from measurement names. Notably, FCS 3.2 supports mixed data types to, for example, allow FCS measurements that are intrinsically integers (e.g., indices or class assignments) or measurements that are commonly captured as integers (e.g., time ticks) to be more represented as integer values, while capturing other measurements as floating-point values in the same FCS data set. These changes are incompatible with existing FCS file readers. The standard retains the overall FCS file structure and most features of previous versions, but also contains a few changes that were required to support new types of data and use cases efficiently. The unchanged goal of the standard is to provide a uniform file format that allows files created by one type of acquisition hardware and software to be analyzed by any other type. FCS 3.2 is a revision of the flow cytometry data standard based on a decade of suggested improvements from the community as well as industry needs to capture instrument conditions and measurement features more precisely.
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