A novel family of intracellular imaging display dyes for measurement of in vitro properties
The University of Edinburgh has created a new family of fluorescein-based dyes in the visible range. They provide end users with the ability to measure intracellular pH, viscosity and temperature.
Measuring the effect of external parameters (eg. drugs) or internal cellular changes (eg. aberrant gene expression) on multiple intracellular properties is of interest to most biologists. This new family of fluorescein analogues have improved stability and can measure multiple intracellular properties. Anthofluorescein has a highly sensitive absorption and fluorescence emission in the pH range from 7 to 10 (see figure) and is proven to have the necessary sensitivity required for optical measurements of intracellular pH inside living cells.
The versatility of the analogues allows the properties of the compounds to be optimised for measurement of other intracellular properties such as temperature and viscosity as well as specific targeting of intracellular organelles. A mitochondrial tag derivative has confirmed that the dyes can be used to successfully label intracellular organelles of choice.
A diacetylated derivative of Anthofluorescein has been demonstrated to be cell permeable and to successfully label living cells. This derivative has applications for cell phenotyping (cancer cells are more acidic), pathology and confocal imaging.
The compound family could be extended to cover the entire spectrum of visible range dyes from yellow to far-red and to allow rapid labelling of bio-molecules (antibodies, proteins, DNA, etc).
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- Visible pH dependent colour change in physiological range (pH7-10)
- In cells, dyes have low extracellular back-ground, with excellent signal-to-noise ratio
- Uniquely multifunctional probes also capable of measuring temperature and viscosity and specific organelle tagging
- Simple synthetic method for the development of further multicolour fluorescence dyes from yellow to far-red
- Studies of intracellular biological systems by tagging specific organelles with a different dye
- Visual imaging of cellular and tissue samples with applications for phenotyping and pathology (eg. cancer / necrotic cells)
- Tools to measure intracellular properties such as pH, temperature and viscosity
- Bio-molecule labelling system with customised colour
Unciti-Broceta et al. "A fluorescein-derived anthocyanidin-inspired pH sensor" Tetrahedron Letters (2009) 50: 3713-3715
Han and Burgess. "Fluorescent indicators for intracellular pH" Chemical Reviews (2009) doi: 10.1021/cr900249z
Three different compounds are currently available:
- Mitochondrial tag that stains the mitochondria that can be used in co-localisation studies
- pH sensor that stains cells
- A diacetylated derivative that is cell permeable and stains all intracellular organelles
The University of Edinburgh is looking to license the fluorophores to a company to manufacture and sell.