How do I claim compensation for FACSDiva?
Select Experiment > Compensation Setup > Calculate Compensation. When the Single Stained Setup box appears, select “Link & Save” to apply the calculated values to your experiment. Load a tube of water on to the instrument and place it in STANDBY. 16.
How do you set up a flow cytometry experiment?
Take a deep breath and begin.
- Double-check the necessary controls. Flow cytometry experiments require a large number of controls for successful interpretation.
- Count your cells.
- Make sure you have your cheat sheets.
- Arrive on time, if not a bit early.
- Annotate your data.
- Follow the protocols.
How do you use compensation beads?
In a 5 mL flow tube, add the same volume of your antibody as you use for your stain. Add one drop of positive comp beads and one drop of negative comp beads, or one drop of the all-in-one comp beads.
How many cells should I use in flow cytometry?
Cell Concentration/Cell number: For each sample, you will need between 10^5 and 10^6 cells. If you are new to flow cytometry, use the higher number of cells — to give yourself a margin for error (you always lose more cells than you expect during the staining and washing procedures).
How many events do you need for flow cytometry?
It is limited to a frequency of 1 in 100,000 beyond which only rough estimates can be obtained even after collecting 10 7 events.
What is Laser delay?
This delay represents the amount of time it takes a cell to move from one laser to the next laser in the relay (see Figure2,right image) and it allows the signals collected from that cell after excitation by all of the lasers in the interrogation point to be delivered to the electronics system all together.
How do you stain comp beads?
Incubate at 2-8°C for 15-30 minutes in the dark. Add 2 mL of Flow Cytometry Staining Buffer to each tube and centrifuge at 400-600 x g for 3-5 minutes. Decant supernatant and add 0.2-0.4 mL of Flow Cytometry Staining Buffer to each tube. Mix briefly by flicking or pulse vortexing before analysis.
How do TruCount beads work?
TruCount absolute-count tubes contain a lyophilized pellet that dissolves during sample preparation, releasing a known number of fluorescent beads. By gating the bead population during analysis, absolute cell counts can be readily determined by a simple calculation.
How do flow cytometers work?
Flow cytometers utilize lasers as light sources to produce both scattered and fluorescent light signals that are read by detectors such as photodiodes or photomultiplier tubes. These signals are converted into electronic signals that are analyzed by a computer and written to a standardized format (. fcs) data file.
What is facsdiva™ software?
BD FACSDiva™ Software provides a graphic of the cytometer’s physical optical configuration, showing the layout of detectors, filters and mirrors. This allows users to select the correct configuration for their work.
Why choose BD facsdiva™ software?
A single daily setup run using BD FACSDiva™ Software and BD FACSDiva™ CS Research Beads* supports consistency and accuracy of data acquisition. Using the Experiment Layout feature in BD FACSDiva™ Software greatly simplifies and speeds up creating a new experiment.
Can I use facsdiva software for in vitro diagnostic (IVD)?
BD FACSDiva software is for in vitro diagnostic (IVD) use when used with IVD reagents and instruments. Refer to the information supplied by the manufacturer for application-specific limitations.
Who is the owner of facsdiva?
BD FACSDiva software © Becton, Dickinson and Company. This software is the property of Becton, Dickinson and Company. Each sale of a stored unit of this software grants the purchaser a nontransferable, nonexclusive, personal license.