Biostatus Ltd Intracellular Dyes, Fluorescence Molecular Imaging Systems
Biostatus Limited was founded in 2001 as a spin-out company from UK academic research. Based in Shepshed, UK, it sells and distributes its products worldwide directly and through carefully selected distributors. Its customers include the pharmaceutical companies, CRO’s, Biotechs and the biomedical research community. Its expertise lies in the development of novel technologies offering solutions for high content screening, microscopy and flow cytometry in the drug discovery and development process. This is backed by intensive technical support. Its core products simplify sample preparation and improve sample analysis in fluorescence based imaging and cytometry, offering higher resolution and more information.
Intracellular Imaging Dyes
DRAQ5 and CyTRAK Orange intracellular imaging dyes permit the rapid and simple “painting” of live and fixed cells. The staining intensity differentiates between the nucleus and cytoplasm allowing image analysis software to segment these for better measurements of drug-related perturbations of cell signalling events such as translocations and phosphorylations, while offering morphometric data that can reflect compound toxicity. They can be easilycombined with fluorescent protein tags such as GFP and permit simultaneous image acquisition. Their ability to enter live cells means that it is possible to study samples without loss of important information due to lysis reagents, fixatives and the excessive washing steps these often entail.
Flow Cytometry Gating
The live cell permeance of DRAQ5 and CyTRAK Orange, along with their high specificity for DNA and excellent spectral properties make them ideal for the analysis of blood and bone marrow using flow cytometry gating strategies, for example in the study of hematotoxicity.
Nucleated cells are then easily gated without the need for time consuming and damaging lysis steps to remove red blood cells, that can also cause unwanted cell aggregation. Analysis can be performed with the minimum of washing steps. The stoichiometric nature of the binding by DRAQ5 provides DNA content measurements for cell cycle analysis and aneuploidy where this is important.
DRAQ5 has become a reagent of choice in microscopy, high content screening and flow cytometry due to its excellent spectral properties. It excites optimally in the red region where there is less problem with biofluorescence and emits in the far-red. DRAQ5 spectra are well separated from all the commonly used fluorescent proteins and fluorescent labels and functional dyes that are used in cell based assays, immunophenotyping, toxicity and cell health measurements. Remarkably, DRAQ5 can be excited by a blue laser and this can allow 4 or 5 colour experiments on simple flow cytometers. Its excellent photo- and chemical-stability mean it can be used in combined techniques such as IF with FISH.
Bone Marrow Differential
The bone marrow differential is an emerging technique to study the toxic effect of drugs on the cellular make-up of the bone marrow compartment. This can also be applied to peripheral blood and lymph node biopsy. DRAQ5 has been demonstrated to simplify and improve the performance of these assays. The use of this live cell far-red DNA dye reduces the complexity of the assay, by removing lysis and washing steps and avoiding the unwanted lysis of late erythroblast precursors which otherwise skews the M:E ratio. The far-red spectral properties make it easy to get many more parameters from a simple one or two laser flow Cytometer.
Fluorescence Molecular Imaging Systems
DRAQ5, DRAQ7 and CyTRAK Orange are compatible with virtually all modern fluorescence molecular imaging systems and flow cytometry systems. Full spectral properties are detailed on the Biostatus website and are exemplified in a wide range of applications across different cell types, tissues and organisms for analytical techniques used in drug discovery and development.
Live Object Immobilization
CyGEL is a thermo-reversible mountant that enables live object immobilization: of non-adherent living cells, tissues and organisms for high-resolution imaging. This has proved a challenge to drug discovery when using hemotopoietic and other suspension cells such as cancer cells and parasites and with model organisms such as zebrafish and C elegans. This hydrogel makes it easy to both immobilize for imaging and injection and then gently recover the live objects for further orthogonal analysis or growth. It is optically clear and inert with little auto-flourescence and a refractive index equivalent to water. Development for HCS applications are currently underway and collaborations are being sought.