I am seeing high background outside of my neuronal cells when using membrane potential indicators. What can I do to reduce background?
If you use our FluoVolt Membrane Potential Kit (Cat. No. F10488), the kit provides a background suppressor to reduce this problem. For other indicators, consider the use of BackDrop Background Suppressor (Cat no. R37603, B10511, and B10512).
What is the difference between fast and slow-response membrane potential probes?
Molecules that change their structure in response to the surrounding electric field can function as fast-response probes for the detection of transient (millisecond) potential changes. Slow-response dyes function by entering depolarized cells and binding to proteins or membranes. Increased depolarization results in additional dye influx and an increase in fluorescence, while hyperpolarization is indicated by a decrease in fluorescence. Fast-response probes are commonly used to image electrical activity from intact heart tissues or measure membrane potential changes in response to pharmacological stimuli. Slow-responding probes are often used to explore mitochondrial function and cell viability.
'The submucous plexus of the guinea pig intestine is a quasi-two-dimensional mammalian neural network that is particularly amenable to study using multiple site optical recording of transmembrane voltage (MSORTV) [Biol. Bull. 183 (1992) 344; J. Neurosci. 19 (1999) 3073]. For several years the potentiometric dye of choice for monitoring the ... More
Cholesterol-enriched lipid domains can be visualized by di-4-ANEPPDHQ with linear and nonlinear optics.
Authors:Jin L, Millard AC, Wuskell JP, Clark HA, Loew LM
Journal:Biophys J
PubMed ID:15879475
We present a membrane-staining dye, di-4-ANEPPDHQ, which differentiates liquid-ordered phases from liquid-disordered phases coexisting in model membranes under both linear and nonlinear microscopies. The dye's fluorescence emission spectrum is blue-shifted 60 nm in liquid-ordered phases compared with liquid-disordered phases, and shows strong second harmonic generation in the liquid-disordered phase compared ... More
Spectral characterization of the voltage-sensitive dye di-4-ANEPPDHQ applied to probing live primary and immortalized neurons.
Authors:Wang Y, Jing G, Perry S, Bartoli F, Tatic-Lucic S,
Journal:Opt Express
PubMed ID:19158915
Spectral properties of a recently developed voltage-sensitive dye, di-4-ANEPPDHQ, were characterized as the dye was dissolved in the solvent dimethyl sulfoxide as the stock solution, in Hank's buffered salt solution as the staining solution, and bound to the plasma membrane of primary rat hippocampal neurons and immortalized mouse hypothalamic neurons ... More
Laurdan and di-4-ANEPPDHQ do not respond to membrane-inserted peptides and are good probes for lipid packing.
Authors:Dinic J, Biverståhl H, Mäler L, Parmryd I,
Journal:Biochim Biophys Acta
PubMed ID:20937246
Laurdan and di-4-ANEPPDHQ are used as probes for membrane order, with a blue shift in emission for membranes in liquid-ordered (lo) phase relative to membranes in liquid-disordered (ld) phase. Their use as membrane order probes requires that their spectral shifts are unaffected by membrane proteins, which we have examined by ... More
Characterization and application of a new optical probe for membrane lipid domains.
Authors:Jin L, Millard AC, Wuskell JP, Dong X, Wu D, Clark HA, Loew LM
Journal:Biophys J
PubMed ID:16415047
In this article, we characterize the fluorescence of an environmentally sensitive probe for lipid membranes, di-4-ANEPPDHQ. In large unilamellar lipid vesicles (LUVs), its emission spectrum shifts up to 30 nm to the blue with increasing cholesterol concentration. Independently, it displays a comparable blue shift in liquid-ordered relative to liquid-disordered phases. ... More