Confocal Microscopy live cell imaging
Why is DRAQ5™ the live cell nuclear stain of choice in confocal microscopy?
- Full spectral compatibility with most fluorescent proteins (e.g. GFP, RFP, etc.)
- No need for UV excitation cf. Hoechst / DAPI
- See the cell architecture, coincident with histone on chromatin
DRAQ5™ can be used on:
- Live cells - primary or cell line-derived
- Live tissue - e.g. drosophila embryo, brain ... see an example here
- Fixed cells/nuclei - methanol, ethanol, paraformaldehyde ....
- Permeabilised cells - by surfactant or naturally in apoptosis
- Tissue sections - frozen or paraffin-embedded
DRAQ5™ IS MULTI-PLATFORM COMPATIBLE
DRAQ5™'s excitation profile is compatible with live cell imaging platforms
- Confocal microscopes - see examples of DRAQ5™ at the manufacturers' websites: e.g. Olympus, Zeiss
- HCS Cell Imaging Systems - DRAQ5™ has been widely used in HCS platforms Opera™: PerkinElmer Evotec Technologies, ArrayScan™ VTI: Cellomics Inc, ImageXpressMICRO: Molecular Devices Corp. (see p.8 of document) IN CELL Analyser: GE Healthcare Biosciences and IN CELL Analyser: GE Healthcare BioSciences & Pathway: BD Biosciences . See more details on cell-based HCS assays in Applications>HTS/HCS
LABELLING INTACT LIVE CELLS:
- Cells can be stained directly without any fixation or permeabilization
- Just a few minutes (and no washing) cells are ready for imaging (protocols)
- Add it to the assay medium in a live cell assay (e.g. GPCR ligands)
Examples from the literature:
Foley et al. Frontiers in Bioscience 2005; 10:1302-1312
Live cell imaging of glial cells & VSMCs was required as both ethanol and PFA fixation generated unwanted artefacts. DRAQ5 was completely compatible with the fluorescein-labelled peptide.
Schjetne et al. The Journal of Immunology 2003; 171:32–36
Live cell imaging was required as they were observing the internalisation of an (Alexa 546) antibody to TLR-2 into cells transfected with GFP-tagged marker of endosomes. DRAQ5, GFP and Alexa 546 were spectrally compatible.
See a review of a paper by Martin et al (2005) for an independent view
See the comparison chart of different live cell dyes based on this paper
FIXED CELLS & NUCLEI
Fix and wash your cells then just add DRAQ5™ in the normal way
Link to the specific section for more information and examples .....
- Email: enquiry@biostatus.com
- Telephone: +44 1509 558163
- Fax: +44 1509 651061