Frequently Asked Questions Regarding Flow Cytometry
To support our customers in the use of products from our flow cytometry portfolio, we have collected the most frequently asked questions with related answers.
If your query is not included in the list below, please do not hesitate to
Q: Can Epstein-Barr Virus (EBER) PNA Probe/Fluorescein, Code Y5200, be used in flow cytometry as described in the package insert? And do you have a protocol for this?A: Yes, Epstein-Barr Virus (EBER) PNA Probe/Fluorescein, Code Y5200, can be used in flow cytometry for RUO purposes. A protocol for the usage on flow cytometry can be found here.
Q: Can I measure MESF (Molecules of Equivalent Soluble Flourochrome) with FluoroSpheres, Code K0110?
A: The beads from the FluoroSpheres, Code K0110, cannot be used for standardization of the fluorescent signal in MESF (Molecules of Equivalent Soluble Flourochrome) units but only MEF (molecules of equivalent flourochrome) values. MEF are used for beads embedded with dyes or flourochromes different from the flourochromes normally used in flow cytometry and thus have different spectral properties.
MESF are used for beads embedded with dyes or flourochromes that are spectrally matched with the flourochromes normally used in flow cytometry.
Q: Can I use another cell line than cell line1301 as control cell line in Telomere PNA Kit/FITC, Code K5327, for flow cytometry? And where can I get cell line 1301?
A: The cell line 1301 has been chosen because the good characteristics of being tetraploid and having long telomeres make it easier to gate on control cells as well as on test cells.
Other control cells can be chosen but be aware of the cell characteristics needed to be able to gate them and differentiate them from the test cells.
Cell line 1301 is available from the cell bank ECACC, with the ECACC number 01051619.
Q: Can Telomere PNA Kit/FITC, Code K5327, be used on mice cells?A: Yes, Telomere PNA Kit/FITC, Code K5327, can be used on hemapoetic cell from specimens different from human. All vertebrates have the same telomere sequence and therefore the probe from Telomere PNA Kit/FITC, Code K5327 can bind telomeres of all vertebrates.
Q: Can Dako's products for flow cytometry be mixed with flow reagents from other vendors?Yes, they can be tested in the same run together with reagents from other vendors as well as mixed in a cocktail together with flow reagents from other vendors. Be aware that the same fluorochromes from different vendors might have slightly different spectral properties and that optimal compensation settings therefore may be different.
Q: What filters should be used when using Cascade Yellow flow products from Dako?A: A proper filter will be a 530/40 nm filter.
Q: Can I use PerCP products in the same channel as RPE-Cy5?A: Yes, both PerCP and RPE-Cy5 are excited by a 488nm laser and emit light at approximately 670 nm.