With all of the cells in organisms, and cells just wandering around in the environment, scientists always seek new ways to sort them. The results unveil basic and applied value. For example, CFD Research Corporation in Huntsville, Ala., is using ALine’s (Redondo Beach, Calif.) flowcell—a laminated system of micrometer-size channels—to sort particles and cells related to biodefense. In fact, various research needs consistently stimulate new tools for tracking and separating cells, often with some form of flow cytometry.
Accurate results from flow cytometry also depend on the software that runs the experiments and gathers data. As the winner of the 2007 Life Science Industry Award for the best flow cytometry-based, cell biology instruments, BD Biosciences in San Jose, Calif., also offers software that supports its devices. In June, this company announced its BD FACSDiva 6.0. This package automates setup for a variety of BD Biosciences flow cytometry systems, including the BD FACSCanto, BD FACSCanto II, BD LSR II and BD FACSAria. For this new software package, BD Biosciences points out several benefits: simplification of multi-color experiments, reproducible data, and the ability to identify performance issues in the device.
As another example, Invitrogen and ScienceXperts in Palo Alto, Calif., teamed up earlier this year to launch CytoGenie. This software helps scientists with the design of flow cytometry experiments, including reagent selection, and it provides a direct link to Invitrogen’s selection of fluorescent dyes. CytoGenie also includes specific protocols for multi-color experiments. Moreover, this package is freely available at: www.invitrogen.com/flowcytometry.
Sometimes the software just needs to help researchers communicate with an instrument. For that, Beckman Coulter recently released a device driver for its Data Innovations Instrument Manager 8.05. This driver controls the information to and from the FC 550 Flow Cytometer.
Part of the excitement in this field comes from combining flow cytometry with other measurements that track cell biology. Earlier this year, for instance, Beckman Coulter in Fullerton, Calif., introduced two new reagents for signal transduction, Phospho-MAPKAPK-2 and Phospho-STAT3 kinase antibodies. These single-color reagents for flow cytometry allow scientists to measure kinase activation in a cell’s cytoplasm, and do it cell by cell. Beckman Coulter points out that these reagents can be used to study to surface-to-nucleus, signal transduction pathways. As examples, Phospho-MAPKAPK2 helps researchers study the mechanism behind inflammation, and Phospho-STAT3 plays a role in apoptosis. Moreover, these reagents even work in complex samples, including blood and bone marrow.
Companies also develop new approaches to sorting. For example, Invitrogen’s (Carlsbad, Calif.) new Dynabeads FlowComp kits can be used in magnetic cell isolation. These polymer-coated beads participate in the isolation of cells, but do not end up in the final product, which just consists of cells in suspension. According to Invitrogen literature, this offers several advantages: minimal interaction with the cells during the process, no need for passing cells through dense columns, and no immunogens remain in the isolated cells. Moreover, Invitrogen ensures that the isolated cells remain functional and ready for downstream assays. In fact, company data show that more than 80% of the cells come out alive, as opposed to less than 50% for some column-based isolation techniques.
Other combination technologies involve the detection instruments. Amnis Corporation in Seattle, Wash., for instance, makes the ImageStream, which provides multispectral-imaging flow cytometry. This system offers many capabilities, from measuring the quantity and location of molecules on cells to virtual cell sorting. In August, Amnis announced two upgrades for this system. First, a multilaser upgrade brought on the potential to add 405 and 658 nanometers lasers—additions to the original 488nm laser already on board in this instrument. This laser upgrade expands the range of dyes that can be used, and it also provides chromatic correction of the dyes excited by the 405nm laser. In addition, an expanded depth of field (EDF) upgrade came out in beta testing. Amnis notes that this increases the device’s depth of field by 10-fold.
The combination of new reagents, labels, and visualization tools gives scientists a better look at more-natural cells after sorting.
About the Author
May is a publishing consultant for science and technology based in Minnesota.
This article was published in Drug Discovery & Development magazine: Vol. 10, No. 11, November, 2007, pp. 14.
Filed Under: Drug Discovery