Data Sheet for
CEN's
Cytometry Control
Product Number 1006
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CEN'sCytometry ControlProduct Number 1006 |
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Chicken Erythrocyte Nuclei (2.0 mL) | |
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BioSure® Chicken Erythrocyte Nuclei (CEN's) are an internal biological reference material that can function as a performance standard when calibrating and troubleshooting flow cytometers. This control can be used as a reference for all types of cells. CEN's are isolated from fresh chicken blood and fixed in ethanol-PBS. The procedure yields single nuclei, doublets, triplets and a few larger aggregates. DNA linear scale histograms of propidium iodide (PI) stained nuclei exhibit multiple peaks; single nuclei constitute peak one (nearest the ordinate) while the next three peaks to the right have mean channel values which are approximately 2, 3, and 4 times greater than the single nuclei peak. A properly aligned flow cytometer will also have a CV for the single nuclei peak of less than or equal to 3%. The published genome size is 2.5 picograms (1).
BioSure® CEN's are supplied as an unstained suspension in 2.0 milliliter volumes. The nuclei count per milliliter is at least 2.0 X 10 to the seventh.
CEN's are stable at 2 - 8 degrees C until the expiration date denoted on each vial label.
To 48 milliliters of calcium and magnesium free Dulbecco's PBS, add 2.5 mg of propidium iodide (PI) and 0.3 mL of Nonidet P40 (NP40). Mix the solution and Q.S. to 50 mL. Staining solution is stable at 2 - 8 degrees C for at least 30 days when stored protected from light in an amber bottle.
The recommended working solution is a 1:10 dilution of the stock CEN's in the PI stain prepared previously. This provides an adequate density of nuclei and sufficient working solution for several hours. 1) Gently mix the CEN stock for 5 - 10 seconds to suspend the nuclei, and add one or two drops to a test tube (13 X 75 mm). 2) Add 1.0 mL of PI stain, mix for 5 - 10 seconds, and incubate at room temperature (22 - 25 degrees C) for 10 minutes protected from light. 3) Mix sample 5 - 10 seconds and evaluate. Samples are typically stable at 2 - 8 degrees C for 4 hours when protected from light. |
Typical CEN graph |
NOTE: The flow cell of some cytometers is susceptible to clogging. Filtering the stained preparation through a 30 - 50 mesh nylon screen will eliminate large nuclear aggregates.
1) Gregory, T.R.; Animal Genome Size Database: 2001-2005, available at http://www.genomesize.com.
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Chicken Erythrocyte Nuclei (2.0 mL) |
Click here for the BioSure® CEN's Alignment and Linearity procedure
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