Voltage-sensitive dye

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Voltage-sensitive dyes, also known as potentiometric dyes, are dyes which change their spectral properties in response to voltage changes. [1] They are able to provide linear measurements of firing activity of single neurons, large neuronal populations or activity of myocytes. Many physiological processes are accompanied by changes in cell membrane potential which can be detected with voltage sensitive dyes. Measurements may indicate the site of action potential origin, and measurements of action potential velocity and direction may be obtained. [2]

Contents

Potentiometric dyes are used to monitor the electrical activity inside cell organelles where it is not possible to insert an electrode, such as the mitochondria and dendritic spine. This technology is especially powerful for the study of patterns of activity in complex multicellular preparations. It also makes possible the measurement of spatial and temporal variations in membrane potential along the surface of single cells.

Types of dyes

Pioneers of voltage-sensitive dyes: A. Grinvald, L.B. Cohen, K. Kamino, B.M. Salzberg, W.N. Ross; Tokyo, 2000 Pioneers of optical recording 2000.jpg
Pioneers of voltage-sensitive dyes: A. Grinvald, L.B. Cohen, K. Kamino, B.M. Salzberg, W.N. Ross; Tokyo, 2000

Fast-response probes: These are amphiphilic membrane staining dyes which usually have a pair of hydrocarbon chains acting as membrane anchors and a hydrophilic group which aligns the chromophore perpendicular to the membrane/aqueous interface. The chromophore is believed to undergo a large electronic charge shift as a result of excitation from the ground to the excited state and this underlies the putative electrochromic mechanism for the sensitivity of these dyes to membrane potential. This molecule (dye) intercalates among the lipophilic part of biological membranes. This orientation assures that the excitation induced charge redistribution will occur parallel to the electric field within the membrane. A change in the voltage across the membrane will therefore cause a spectral shift resulting from a direct interaction between the field and the ground and excited state dipole moments.

New voltage dyes can sense voltage with high speed and sensitivity using photoinduced electron transfer (PeT) through a conjugated molecular wire. [3] [4]

Slow-response probes: These exhibit potential-dependent changes in their transmembrane distribution which are accompanied by a fluorescence change. Typical slow-response probes include cationic carbocyanines and rhodamines, and ionic oxonols.

Examples

Commonly used voltage sensitive dyes are substituted aminonaphthylethenylpyridinium (ANEP) dyes, such as di-4-ANEPPS, di-8-ANEPPS, and RH237. Depending on their chemical modifications which change their physical properties they are used for different experimental procedures. [5] They were first described in 1985 by the research group of Leslie Loew. [6] ANNINE-6plus is a voltage sensitive dye with fast response (ns response time) and high sensitivity. It has been applied to measure the action potentials of a single t-tubule of cardiomyocytes by Guixue Bu et al. [7] More recently, a series of fluorinated ANEP dyes was introduced that offer enhanced sensitivity and photostability; they are also available over a wide choice of excitation and emission wavelengths. [8] A recent computational study confirmed that the ANEP dyes are affected only by the electrostatic environment and not by specific molecular interactions. [9] Other structural scaffolds, such as xanthenes, [10] are also successfully used.

Materials

The core material for imaging brain activity with voltage-sensitive dyes are the dyes themselves. These voltage-sensitive dyes are lipophilic and preferably localized in membranes with their hydrophobic tails. They are used in applications involving fluorescence or absorption; they are fast acting and are able to provide linear measurements of changes in membrane potential. [11] Voltage sensitive dyes are supplied by many companies who offer fluorescent probes for biological applications. Potentiometric Probes, LLC specializes only in voltage sensitive dyes; they have an exclusive license to distribute the large set of fluorinated VSDs, marketed under the ElectroFluor brand.

A variety of specialized equipment may be used in conjunction with the dyes, and choices in equipment will vary according to the particularities of a preparation. Essentially, equipment will include specialized microscopes and imaging devices, and may include technical lamps or lasers. [11]

Strengths and weaknesses

Strengths of imaging brain activity with voltage-sensitive dyes include the following abilities:

Weaknesses of imaging brain activity with voltage-sensitive dyes include the following problems:

Uses

Voltage-sensitive dyes have been used to measure neural activity in several areas of the nervous system in a variety of organisms, including the squid giant axon, [19] whisker barrels of the rat somatosensory cortex, [20] [21] olfactory bulb of the salamander, [22] [23] [24] visual cortex of the cat, [25] optic tectum of the frog, [26] and the visual cortex of the rhesus monkey. [27] [28]

Many applications in cardiac electrophysiology have been published, including ex vivo mapping of electrical activity in whole hearts from various animal species, [29] [30] subcellular imaging from single cardiomyocytes, [31] and even mapping both sinus rhythms and arrhytmias in open heart in vivo pig, [18] where motion artifacts could be eliminated by dual wavelength ratio imaging of the voltage sensitive dye fluorescence.

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<span class="mw-page-title-main">Electrophysiology</span> Study of the electrical properties of biological cells and tissues.

Electrophysiology is the branch of physiology that studies the electrical properties of biological cells and tissues. It involves measurements of voltage changes or electric current or manipulations on a wide variety of scales from single ion channel proteins to whole organs like the heart. In neuroscience, it includes measurements of the electrical activity of neurons, and, in particular, action potential activity. Recordings of large-scale electric signals from the nervous system, such as electroencephalography, may also be referred to as electrophysiological recordings. They are useful for electrodiagnosis and monitoring.

<span class="mw-page-title-main">Action potential</span> Neuron communication by electric impulses

An action potential occurs when the membrane potential of a specific cell rapidly rises and falls. This depolarization then causes adjacent locations to similarly depolarize. Action potentials occur in several types of animal cells, called excitable cells, which include neurons, muscle cells, and in some plant cells. Certain endocrine cells such as pancreatic beta cells, and certain cells of the anterior pituitary gland are also excitable cells.

<span class="mw-page-title-main">Dendritic spine</span> Small protrusion on a dendrite that receives input from a single axon

A dendritic spine is a small membranous protrusion from a neuron's dendrite that typically receives input from a single axon at the synapse. Dendritic spines serve as a storage site for synaptic strength and help transmit electrical signals to the neuron's cell body. Most spines have a bulbous head, and a thin neck that connects the head of the spine to the shaft of the dendrite. The dendrites of a single neuron can contain hundreds to thousands of spines. In addition to spines providing an anatomical substrate for memory storage and synaptic transmission, they may also serve to increase the number of possible contacts between neurons. It has also been suggested that changes in the activity of neurons have a positive effect on spine morphology.

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Further reading