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1. #Flowjo reference how to#
2. #Flowjo reference manual#
3. #Flowjo reference software#
4. #Flowjo reference series#

#Flowjo reference software#

This includes technologies deployed as software installations on VMs within VA-controlled cloud environments (e.g. The TRM decisions in this entry only apply to technologies and versions owned, operated, managed, patched, and version-controlled by VA. This technology stores data using a proprietary database. It is used for testing Deoxyribonucleic Acid (DNA)/Cell Cycle, Kinetics, and Proliferation, and works with most Fluorescence Activated Cell Sorting (FACS) machines and data hosting services (Laboratory Information Management Systems). The program allows users to organize and manage experiments and produce reports. More information on the proper use of the TRM can be found on theįlowJo is a software package used for viewing and analyzing flow cytometry data.

#Flowjo reference series#

Examine these two series of pages to see how easy it is to calibrate your samples with FlowJo!ĭownload a Calibration Workspace with Demo Data to try out this platform.Technologies must be operated and maintained in accordance with Federal and Department security and Both use reference samples one uses a bead set, the other, the expression of CD4 on lymphocytes as a defined set point. In addition, you can see two examples of creating Calibration Standards.

#Flowjo reference how to#

To see how to use either method to create a new Calibration Standard, click on this link. It asks you to define which parameter you want to calibrate, and then which method to use. When you select "Calibrate." under the Workspace menu, you will be presented with the " Define New Calibration Standard" dialog window. For more information on derived parameters, click here. They are accessible as Derived parameters thus, if you want to apply a Calibration Standard to another sample (after defining it), select its name under the "Derived Parameters" menu item. Calibration standards are saved with the workspace, permanently. FlowJo then performs a linear (or logarithmic) least squares regression to determine the best calibration value, and creates the calibrated parameterĪ "Calibration standard" is FlowJo's internal representation of the conversion of fluorescence to absolute units. When you use a reference, FlowJo automatically finds all of the peaks in the fluorescence, and asks you to enter any or all of the calibration values for those peaks. Use the reference when you have collected data on a standard which you can use to convert to your units (for example, a commercial bead set where each bead has a defined absolute intensity or, a sample stained with an antibody that has a known binding capacity to the cells).

#Flowjo reference manual#

Use the manual mode when you want to enter the exact conversion values yourself. There are two methods by which you can calibrate a parameter: manual or reference. Finally, you can even enter a gate based on those units: you could, for example, specify a gate on all cells with more than 5,000 molecules of an antigen. In addition, any statistics you calculate (medians, CVs, etc.) will be expressed in units of absolute molecules. When you graph your data, select the calibrated parameter on the axis of choice-the scale values on the axis are now drawn in your calibrated units! For example, if you calibrate to absolute number of molecules per cell, then you can display these numbers on the axis. You may have as many different calibration standards in a workspace as you want in fact, you can even apply multiple calibration standards to the same sample (a new parameter will be added for each calibration that you apply). You can apply the calibration standard to any other sample in your collection (and you can apply the calibration to an entire group, if you wish). Once you have created a calibrated parameter, it is added to your sample as if it were just another parameter. Most commonly, this is used to convert the scaling into absolute number of molecules (given a standard that converts between the fluorescence intensity collected on your instrument and absolute numbers of fluorophores). This gives you the ability to express any parameter's values (graphical display, any statistical calculations) in the units of your choice. FlowJo has a unique platform that allows you to calibrate any collected parameter.