How do compensation beads work?

How do compensation beads work?

Compensation beads are small particles (typically polystyrene) that are pre-coated with antibodies recognizing species-specific antibody light chains. Special care must be taken to ensure that the compensation beads you choose will bind to the species in which your fluorochrome- conjugated antibody was raised.

What is a bead in flow cytometry?

Compensation beads capture species-specific antibodies conjugated to fluorophores and other types of reagents. The purpose of these beads is to set voltages and gating parameters for obtaining accurate fluorescence signal.

How do you use ultra comp beads?

Experimental procedure

  1. Label a tube for each fluorochrome that will be used in the experiment.
  2. Mix beads by vigorously inverting at least 10 times or pulse-vortexing.
  3. Label each tube and pulse vortex 10 times.
  4. Add 1 drop of UltraComp eBeads to each tube.
  5. Add 1 test or less of antibody conjugate to each tube and mix.

Why are beads used in flow cytometry?

These beads provide: Consistent, accurate, and simple-to-use reagents for setting flow cytometry compensation when using green fluorescent protein (GFP). Ease-of-use with a single drop containing both negative and positive beads.

What is compensation flow?

UNIT 1.14. Compensation in Flow Cytometry. The term “compensation,” as it applies to flow cytometric analysis, refers to the process of correcting for fluorescence spillover, i.e., removing the signal of any given fluorochrome from all detectors except the one devoted to measuring that dye.

How do you use AccuCheck counting beads?

Mix and incubate in the dark for 10minutes at room temperature. Add 1 ml of deionized water (kept at room temperature), incubate for 5 minutes and vortex*. A. 4 Immediately prior to use, carefully mix AccuCheck Counting Beads for 30-45 seconds manually (do not vortex).

What are CS beads?

BD CS Beads are a suspension of fluorospheres with uniform and stable size and fluorescence intensity. The beads are used for instrument quality control (QC) to characterize, track, and report performance measurements of supported flow cytometers. The cytometer’s software displays current bead data in plots.

How do counting beads work flow cytometry?

The formula has you divide the number of cells in the region you want to count by the number of beads analyzed. This value is then multiplied by the number of beads you added.

How is flow compensation calculated?

Altogether, the flow compensated can be calculated with actual temperature, actual pressure, and actual molecular weight, etc. Molecular weight can be known from Lab analysis or online analyzer. Pressure values shall be in Bar(a), Temp shall be in K.