Luminex Technology Multiplex Assays

We continue to support customers of our pre-existing (non-ProcartaPlex) portfolio of Invitrogen assays for the Luminex platform. Contact us at LuminexFAS@thermofisher.com for personalized assistance. While supplies last, selected legacy singleplex kits and multiplex panels remain available for purchase. Navigate to the following product categories, then search or filter results by the word “Novex”.

• Singleplex Luminex Technology Protein Assays
• Multiplex Luminex Technology Protein Assays

xMAP Technology

The xMAP technology applies to two bead types—magnetic and polystyrene. The magnetic Magplex bead is a superparamagnetic 6.5 micron microsphere with a magnetic core and polystyrene surface. The polystyrene Microplex bead is a 5.6 micron polystyrene microsphere. Both bead types are internally dyed with precise proportions of red and infrared fluorophores. The differing proportions of the red and infrared fluorophores result in 100 unique spectral signature microspheres which are identified by the Luminex xMAP detection systems. The conjugation of a distinct monoclonal antibody to a distinct bead allows for analysis of multiple analytes in a single well.

The polystyrene bead requires expensive, and sometimes difficult to use filter plates. This bead type also makes automating the assay more difficult. The magnetic bead provides significant advantages over the polystyrene bead because it allows for greater ease and flexibility. This ease is realized in the ability to use a flat-bottom plate with either a hand-held magnetic plate washer (cat. no. EPX-55555-00) or an automated magnetic plate washer. In addition, a filter plate/vacuum manifold system, commonly used with polystyrene beads, can also be used with magnetic beads.

Luminex Instrumentation

Luminex instrumentation is comprised of three instrument models—MAGPIX, LX200 and FLEXMAP 3D. These instruments are based on either the principles of quantitative fluorescent microscopy or fluorescent flow cytometry. The multiplex quantitation capability of an assay is dependent upon instrument model, and ranges from 50 analytes with use of the MAGPIX, to 500 analytes with the FLEXMAP 3D.  Used in combination with ProcartaPlex, the Luminex platform delivers fast and cost-effective immunoassay profiling analysis.

The Luminex System is a series of three instruments that flexible analyzer based on the principles of quantitative fluorescent microscopy or fluorescent flow cytometry that is designed to meet the needs of any size research laboratory. The multiplex system allows for the simultaneously measurement of up to 80 different analytes from a single microplate well, using very small sample volumes. The system delivers fast and cost-effective immunoassay profiling and gene expression analysis through the multiplex quantitative measurement and detection of protein, RNA, and DNA.

The Luminex System is the combination of three core xMAP technologies. The first is xMAP microspheres, a family of 100 fluorescently dyed 5.6 magnetic microspheres (beads) and 5.6 micron-sized polystyrene microspheres. Both bead types act as both the target identifier and the solid surface capture to build the assay. The second is a fluorescent imaging and flow cytometry-based instruments, the Luminex Magpix and Luminex 200/FLEXMAP 3D analyzer, respectively, which integrates key xMAP detection components such as light emitting diodes (LEDs) and a CCD camera or lasers, optics, advanced fluidics and high-speed digital signal processors. The third component is the assays that are designed around the microspheres.

MAGPIX

More recently Luminex introduced the MAGPIX system which is based on the principles of fluorescence imaging. Lasers and Photo Multiplying Tubes (PMTs) were replaced with Light Emitting Diodes (LEDs) and a CCD camera to deliver a cost effective, compact, and reliable multiplexing platform. The compact MAGPIX instrument is capable of simultaneously measuring and quantitating up to 50 analytes in a single microtiter well when used with magnetic micropsheres (beads) (described above). The MAGPIX instrument is ideal for low to medium-throughput labs for the quantitative analysis of proteins, RNA, and DNA in a variety of sample matrices.

Luminex 200 and FLEXMAP 3D

The Luminex 200 and FLEXMAP 3D reader combines two lasers, fluidics, and real-time digital signal processing to distinguish up to 80 different sets of color-coded magnetic microsphere beads, each bearing a different assay. The Luminex reader is an essential tool that performs the key functions of this multiplex technology:

Fluidics

The reader detects individual beads by flow cytometry. The fluidics system of the reader aligns the beads into single file as they enter a stream of sheath fluid and then enter a flow cell. Once the beads are in single file within the flow cell, each bead is individually interrogated for bead color (analyte) and assay signal strength (PE fluorescence intensity).

Lasers

The reader uses a 532 nm green laser ("assay" laser) is used to excite the PE dye of the assay (streptavidin-PE). The 635 nm solid state laser (red "classify" laser) is used to excite the dyes inside the beads to determine their "color" or "region" and is also used for doublet discrimination by light scatter.

Detectors

The reader has four detectors, one for each of the optical paths shown in the figure below. Detectors are used to measure the fluorescence of the assay, to make bead determination (1-100) and the last to discriminate between single and aggregate beads.

Hand-Held magnetic plate washer for magnetic bead assays

We have developed a proprietary hand-held magnetic plate washer (sold separately) to simplify the bead wash process.