Aug 03, 2020
FCMPASS - Importing fcs files
- 1Translataional Nanobiology Section, Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of Health
- Translational Nanobiology Section
Protocol Citation: Joshua A Welsh, Jennifer Jones 2020. FCMPASS - Importing fcs files . protocols.io https://dx.doi.org/10.17504/protocols.io.bhvxj67n
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 25, 2020
Last Modified: June 29, 2023
Protocol Integer ID: 38551
Keywords: fcmpass, flow cytometry, calibration, EVS,
Disclaimer
This protocol summarizes key steps for a specific type of assay, which is one of a collection of assays used for EV analysis in the NCI Translational Nanobiology Section at the time of submission of this protocol. Appropriate use of this protocol requires careful, cohesive integration with other methods for EV production, isolation, and characterization. By using the FCMPASS software you agree to the following terms and conditions.
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Abstract
This protocol outlines the steps required to import fcs files for calibration using the FCMPASS software. This is one of a number of protocols in the pipeline for performing small particle calibration using the fcmpass software package.
Materials
FCMPASS software can be accessed at https://nanopass.ccr.cancer.gov.
Once a dataset has been created click the ‘Begin Calibration’ button.
Import fcs files by selecting the ‘+’ icon next to the ‘Files to calibrate’ table.
In the new window navigate to the folder containing the fcs files you wish to calibrate and select ‘OK’.
The fcs files and related metadata will now be imported.
If the folder contains fcs files that you do not wish to be calibrated, select them and click the ‘–‘icon. The metadata related to the remaining files will then be reprocessed.
The parameters e.g. SSC-H, SSC-A that are available in further steps of the software are those that are common to all the loaded fcs files. If files that are loaded do not have any common parameter names a selection will not be available in these steps.
Under the ‘Sample Type’ column all loaded files by default are listed as ‘Sample’. For the relevant files these can be adjusted to ‘SSC Calibration’ or ‘FL Calibration’ depending on what the sample was used for.
Once completed select ‘Next’.