Bender MedSystems FlowCytomix™ FAQs

Is there a particular flow cytometer required for the use of Bender MedSystems FlowCytomix™ Kits?

FlowCytomix™ microspheres are detectable in most commonly used flow cytometers. The flow cytometer needs to be equipped with one laser (488 nm or 532 nm) capable of detecting and distinguishing fluorescence emissions at 575 nm and far red (685 nm - 690 nm).

Bender MedSystems FlowCytomix™ suitability has been tested for:

Beckman Coulter:
EPICS® XLTM / XL-MCL™*
Cytomics TM FC500*

BD FACScan™
BD FACSCalibur™
BD FACSCanto™, FACSCanto™ II
BD TM LSR I
BD TM LSR II

Guava EasyCyte Plus

MACSQuant® 

* For the most commonly used flow cytometers (e.g. Becton Dickinson or DakoCytomation) you can immediately start cytometer setup as below.

If you run a Bender MedSystems FlowCytomix™ assay on a FC500 Instrument from Beckman Coulter you must ensure that the Forward Scatter (FS) measurements are collected at 1-8°. To accomplish this you must insert the FS 1-8° Field Stop into place: remove the front cover to locate the FS 1-8° Field Stop. Slide the knob from the right (= default 1-19° position) to the left and push to lock in place.

On the XL Series systems the EPICS XL/XL-MCL FS Low Angle Collection Kit is recommended to accomplish this.

Is there a special software required?

Yes, the flow cytometer raw data files are analysed with the specifically designed FlowCytomix Pro software. 

The FlowCytomix™ Pro 2.3 (Cat. No. BMS8401FF) Software is complimentary and can be ordered at europe@ebioscience.com or downloaded at www.bendermedsystems.com/software-download

Does the Bender MedSystems FlowCytomix™ Pro software also run on Apple Macintosh?

It is possible to run FlowCytomix™ Pro software on Apple Macintosh with Mac OS X or higher. For

Does the Bender MedSystems FlowCytomix Pro software interact with the flow cytometer operating system?

The FlowCytomix™ Pro software has been virus checked with the latest version of McAfee VirusScan. The FlowCytomix™ Pro Software has been successfully operated in conjunction with all commonly used flow cytometer models. It does not interfere with the flow cytometer operating system.

In addition the FlowCytomix™ Pro Software does not need to be installed on the flow cytometer working station; data files can be transferred to any other computer.

What happens if less than 11 analytes have to be measured?

FlowCytomix™ Simplex Kits are available for this purpose. FlowCytomix™ Simplex Kits are designed for the detection of one specific cytokine. Simplex Kits can be combined to give any combination (within the human, mouse or rat range) and number of cytokines to be measured.

When is a FlowCytomix™ Basic Kit required?

A Basic Kit is required for any combination of Simplex Kits. In order to run a FlowCytomix™ assay some reagents are needed only once, even in the case of combining several Simplex Kits. These reagents are provided in a Basic Kit.

Are the assays performed in plates or in tubes?

The FlowCytomix™ assays can be either performed like a Bender MedSystems ELISA in a 96-well filter plate. In this case, a filtration manifold is required. Alternatively, the assay can be performed in tubes, which requires centrifugation of the tubes during the washing procedure.

Why is a cytometer setup required before measuring FlowCytomix™ standards and samples?

The cytometer setup is required to prepare the instrument for the Bender MedSystems bead-based assays. FlowCytomix™ setup beads are included in the Basic Kits and in the Multiplex Kits to prepare the correct instrument settings.

With every new experiment, before starting the acquisition of standards and samples, stay in SET UP MODE and adjust the settings using the highest standard (Standard 1).

What wavelength is chosen for measurement?

The maximum emission of PE is at 578 nm and the emission of the beads is in the far red region (685 - 690 nm).

Is it possible to measure with high "flow through" settings?

There are often three flow through settings: low, medium, & high.

Increasing the "flow through" rate may result in a dispersed bead population. Therefore it is recommended to start with the low "flow through". The rate can be increased if the bead population does not start to disperse.

Once the final instrument settings are saved, do not change the "flow through" rate during measurement.

Does a FlowCytomix™ Kit work on a Luminex™ instrument?

No, the FlowCytomix™ Kit can only be measured on a flow cytometer.

Is it correct to use the non linear part of the standard curve?

Yes, it is possible to use the non linear part of the standard curve for calculation of results. Dilutions of samples behave in the same way as the standard curve.

FlowCytomix™ FAQ´s regarding Combination:

How many analytes can be combined in one assay?

With FlowCytomix™ up to 20 analytes are possible.

Which analytes can be combined?

Up to 20 FlowCytomix™ Simplex Kits within one species can be combined.
FlowCytomix™ Simplex Kits with an identical bead population cannot be combined.

For detailed information please refer to Possible Combinations!

Can FlowCytomix™ Multiplex Kits be combined with each other?

All FlowCytomix™ human Multiplex Kits can be combined with BMS810FF human Th1/Th2 11plex. All FlowCytomix™ mouse Multiplex Kits can be combined with BMS820FF mouse Th1/Th2 10plex. A combination of BMS811FF human Cardiovascular Panel with BMS812FF human Adhesion Panel is possible for some analytes which do not use the same bead sets.

Can FlowCytomix™ Simplex Kits be combined with a Multiplex Kit?

Yes, FlowCytomix™ human Simplex Kits can be combined with human Multiplex Kits provided the beads are compatible as described here: Possible Combinations

Can FlowCytomix™ components from different lots be mixed?

The FlowCytomix™ bead sets, standards and conjugates are lot-specific and must be used in combination with each other. Do not mix these components from different kit lots.

FlowCytomix™ Assay Buffer, Reagent Dilution Buffer and Streptavidin-PE are not lot-specific and can therefore be exchanged between different kit lots.

Is it possible to combine FlowCytomix™ Biotin Conjugates with PE-conjugates in one conjugate mixture?

Yes. But please note, that in case you are using at least one Biotin-Conjugate in your combination a further incubation step with Streptavidin-PE is necessary. This does not interfere with the PE-conjugates.