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The Jump Step Histogram Simulator is a Microsoft Excel Spreadsheet that simulates jump step histograms for diffusing particles, including the effect of localization precision. A maximum of three distinct molecular species are allowed in the population, including additional features of jiggle, linear or random drift, and rotational motion of large particles. System Requirements: Microsoft Excel ver. 16.76 (23081101) or later Tested on the above software version; no non-standard hardware required. Installation: Copy/download file to computer and open with Microsoft Excel. Typically takes a few minutes. Demo: Enable ‘Calculation’ within Excel to run a simulation. The normal output is a histogram of jump step sizes and R2/t values for the input parameters. Run time is a few minutes. Instructions: A description of various features and how to input parameters and run the simulation are explained within the Excel file.