Flow cytometry is a powerful technique for simultaneously analyzing multiple characteristics of individual cells within a population in a relatively short period of time. The LSR II from BD is a benchtop flow cytometer that can be configured to detect up to 18 colors or parameters. The LSR II has increased performance capabilities compared to its predecessor, the LSR. This, together with a high degree of flexibility, versatility and ease of use makes the LSR II a state of the art instrument that allows you to do multiparametric analyses on single cells across a wide range of advance research applications, including clinical and basic science.
The LSRII combines up to 4 fixed, aligned, air-cooled lasers which include: Lightwave Xcyte UV laser (20 mW @ 355 nm), a Sapphire blue laser (20 mW @488 nm), a VioFlame Plus violet laser (25 mW @ 405 nm), and a HeNe laser (18 mW @ 633 nm). The revolutionary optical design uses octagon and trigon arrays for increased resolution and flexibility in a fiber optic system. Additionally, signal processing is achieved by a digital acquisition system providing more accurate fluorescent measurements. The electronics are able to digitize 10 million signals per second in 16,384 discrete levels. The LSR II is driven by BD FACSDiva™ software with features such as reusable acquisition templates, automated compensation, and offline compensation.
The BD FACSDiva™ is BD’s new flow cytometry PC-based software which was previously used with other models like the BD FACSVantage™. This software allows researches to quickly and efficiently set up, acquire, and analyze flow cytometry worksheets and includes interesting analysis tools like: Snap-To gating and batch analysis. However, there is still a lack of efficient data export tools. This really diminishes your options for handling your results for presentations and more importantly, for “journal quality” figure creation. Other really useful features that were present in the previous LSR software, BD CellQuest™ (Macintosh based) are now missing, including histogram overlay. So, if you really want to make the most out of the LSR II you will need to get another software package for the plotting and analysis of your data.
We previously used the LSR which we have now upgraded to the LSR II. The LSR II Flow Cytometer is presently used in our lab on a daily basis. Our studies involve mainly multiparametric analysis of apoptosis and immunophenotyping. For the case of apoptosis, where the biochemical events occur with stochastic kinetics, flow cytometry is a great tool that permits us to identify discrete populations of cells within either homogenous or heterogeneous cell preparations. These studies include the analysis of changes in cell morphology (cell volume, granularity, lipid asymmetry), changes in the intracellular milieu (ionic imbalance, redox potential, pH) and other biochemical parameters including the activation signal transduction pathways (kinases, second messengers and caspases). In this way the LSR II has allowed us to simultaneous analyze more than 4 different parameters and design many assays which include:
a) Simultaneous analysis of changes in intracellular glutathione and the generation of reactive oxygen species including superoxide anion and nitric oxide.
b) Simultaneous analysis of mitochondrial and plasma membrane potential together with cell viability.
c) Simultaneous study of changes in cell volume, membrane blebbing and ionic gradients of calcium, potassium and sodium.
d) Simultaneous characterization of changes in phosphatidyl-serine distribution and DNA degradation.
Additionally, other neighbor laboratories within the institute use it for a wide variety of applications including phenotypic analysis, cell cycle and proliferation studies, several functional studies like ion fluxes, changes in pH and the intracellular redox state, cell tracing, activation of signal transduction pathways, Fluorescence Resonance Energy Transfer (FRET) and gene expression.
Overall, the BD LSRII flow cytometer is as good as you can get now days. Users will be surprised with all the advantages this instrument offers that allows you to further reach your research goals.
Rodrigo Franco Cruz, PhD
Visiting Fellow
National Institute of Environmental Health Sciences, NIH
Molecular Endocrinology Group
Laboratory of Signal Transduction