Voltage dependent dye

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A voltage-dependent dye (also potentiometric dye , voltage-sensitive dye ) is a dye that changes its color when a voltage is applied .

properties

Voltage-dependent dyes are used, among other things, in electrophysiology and neurobiology in order to be able to microscopically follow changes in membrane potential such as action potentials , e.g. B. in neurons and myocytes . This shows the origin, the direction and the speed of propagation of the action potential. The change in color is evident in the event of a voltage change through a change in the extinction coefficient or through a shift in the maximum in the absorption spectrum or, in the case of fluorescent dyes, through a shift in the maximum in the emission spectrum . The exact mechanism of the change in color or increase in color of the various types of dye is still unknown. In the case of voltage-dependent merocyanine dyes, there is no shift in wavelength, but only an increase in fluorescence. Therefore, various mechanisms have been discussed, e.g. B. the dislocation of the dye between the membrane and the aqueous environment, a change in the dielectricity of the surrounding biomembrane or dimerizations and their dissociation. In contrast, there is a shift in the spectral maxima with ANEP dyes.

The color change can be followed by a decrease in the extinction coefficient at the original wavelength or by an increase in the extinction coefficient at the wavelength of the changed color. Since voltage-dependent dyes are less precise in determining the change in voltage compared to electrodes , they are mostly used where electrodes cannot be inserted, e.g. B. in mitochondria . Alternatively, voltage-dependent reporter proteins are used, e.g. B. VSFP or PROPS.

Compared to electrodes, many neurons can be observed in parallel, including the direction and speed of the potentials. The process is partially reversible; after observation, the cells can be rinsed with culture medium to remove the dye. The reproducibility , the signal-to-noise ratio and the sensitivity are comparatively lower. The diffusion through connective tissue is less than through other tissues. Colorants can have undesirable pharmacological effects, e.g. B. an increase in photosensitivity . Serum components in the culture medium can reduce the fluorescence, which is why it is usually stained in an isotonic buffer.

Types

  • Merocyanine 540

  • DiBAC 4

  • ANNINE-6

  • ANNINE-6plus

  • DI-4-ANEPPS

  • DI-8-ANEPPS

  • RH237

Slow-response probes change their distribution coefficient after a change in voltage and are only then stored in the cell membrane, e.g. B. some cationic cyanines (e.g. Merocyanin 540), rhodamines , and ionic oxonols (e.g. DiBAC 4 ).

Fast-response probes (in German, fast-reacting probes') are generally amphiphilic dyes having aliphatic side chains in the cell membrane store and a hydrophilic radical with the fluorophore z. B. ANNINE-6, ANNINE-6plus. Voltage dependent dyes are often aminonaphthylethenylpyridine dyes, e.g. B. di-4-ANEPPS (1- (3-sulfonatopropyl) -4- [β- [2- (di-n-butylamino) -6-naphthyl] vinyl] pyridinium betaine), di-8-ANEPPS and RH237.

history

The voltage-dependent dye merocyanine 540 was first published in 1976 by Guy Salama and Martin Morad. The ANEP dyes were described by Leslie Loew's group from 1985 onwards.

literature

  • Leslie M. Loew: Potentiometric dyes: Imaging electrical activity of cell membranes. In: Pure Appl. Chem. (1996), Vol. 68, No. 7, pp. 1405-1409.

Individual evidence

  1. ↑ LD Liao, V. Tsytsarev, I. Delgado-Martínez, ML Li, R. Erzurumlu, A. Vipin, J. Orellana, YR Lin, HY Lai, YY Chen, NV Thakor: Neurovascular coupling: in vivo optical techniques for functional brain imaging. In: Biomedical engineering online. Volume 12, 2013, p. 38, doi : 10.1186 / 1475-925X-12-38 . PMID 23631798 . PMC 3655834 (free full text).
  2. ↑ a b c d e f Baker BJ, Kosmidis EK, Vucinic D, et al. : Imaging brain activity with voltage- and calcium-sensitive dyes . In: Cell. Mol. Neurobiol. . 25, No. 2, March 2005, pp. 245-82. doi : 10.1007 / s10571-005-3059-6 . PMID 16050036 .
  3. ^ Cohen, Lawrence B and Salzberg, Brian M, Optical Measurement of Membrane Potential . In: Reviews of Physiology, Biochemistry and Pharmacology , vol. 83, pp. 35-88, 1978; doi : 10.1007 / 3-540-08907-1_2
  4. ^ A b c G. Salama, BR Choi: Images of Action Potential Propagation in Heart. In: News in physiological sciences: an international journal of physiology produced jointly by the International Union of Physiological Sciences and the American Physiological Society. Volume 15, February 2000, pp. 33-41, PMID 11390873 . PDF .
  5. ↑ David Robinson, Besley, Nicholas A .; O'Shea, Paul; Hirst, Jonathan D .: Di-8-ANEPPS Emission Spectra in Phospholipid / Cholesterol Membranes: A Theoretical Study . In: Journal of Physical Chemistry B . 115, No. 14, April 14, 2011, pp. 4160-4167. doi : 10.1021 / jp1111372 .
  6. ↑ WY Kao, CE Davis, YI Kim, JM Beach: Fluorescence emission spectral shift measurements of membrane potential in single cells. In: Biophysical Journal. Volume 81, Number 2, August 2001, pp. 1163-1170, doi : 10.1016 / S0006-3495 (01) 75773-6 . PMID 11463657 . PMC 1301585 (free full text).
  7. ↑ H. Mutoh, A. Perron, W. Akemann, Y. Iwamoto, T. Knöpfel: Optogenetic monitoring of membrane potentials. In: Experimental Physiology . Volume 96, number 1, January 2011, pp. 13-18, doi : 10.1113 / expphysiol.2010.053942 . PMID 20851856 . PDF .
  8. ↑ H. Mutoh, W. Akemann, T. Knöpfel: Genetically engineered fluorescent voltage reporters. In: ACS Chemical Neuroscience . Volume 3, Number 8, August 2012, pp. 585-592, doi : 10.1021 / cn300041b . PMID 22896802 . PMC 3419450 (free full text).
  9. ↑ JM Kralj, DR Hochbaum, AD Douglass, AE Cohen: Electrical spiking in Escherichia coli probed with a fluorescent voltage-indicating protein. In: Science. Volume 333, Number 6040, July 2011, pp. 345-348, doi : 10.1126 / science.1204763 . PMID 21764748 .
  10. ^ A b G. Salama, M. Morad: Merocyanine 540 as an optical probe of transmembrane electrical activity in the heart. In: Science. Volume 191, Number 4226, February 1976, pp. 485-487, PMID 1082169 .
  11. ^ B. Kuhn, P. Fromherz, W. Denk: High sensitivity of Stark-shift voltage-sensing dyes by one- or two-photon excitation near the red spectral edge. In: Biophysical Journal. Volume 87, Number 1, July 2004, pp. 631-639, doi : 10.1529 / biophysj.104.040477 . PMID 15240496 . PMC 1304385 (free full text).
  12. ↑ P. Fromherz, G. Hubener, B. Kuhn, MJ Hinner: ANNINE-6plus, a voltage-sensitive dye with good solubility, strong membrane binding and high sensitivity. In: European biophysics journal: EBJ. Volume 37, Number 4, April 2008, pp. 509-514, doi : 10.1007 / s00249-007-0210-y . PMID 17687549 . PMC 2755735 (free full text).
  13. ↑ Bu G., et al .: Uniform action potential repolarization within the sarcolemma of in situ ventricular cardiomyocytes. . In: Biophysical Journal . 96, No. 6, March 2009, pp. 2532-2546. bibcode : 2009BpJ .... 96.2532B . doi : 10.1016 / j.bpj.2008.12.3896 . PMID 19289075 . PMC 2907679 (free full text).
  14. ↑ E. Fluhler, VG Burnham, LM Loew: Spectra, membrane binding, and potentiometric responses of new charge shift probes. In: Biochemistry. Volume 24, Number 21, October 1985, pp. 5749-5755, PMID 4084490 .
  15. ^ A. Bullen, P. Saggau: High-speed, random-access fluorescence microscopy: II. Fast quantitative measurements with voltage-sensitive dyes. In: Biophysical Journal. Volume 76, Number 4, April 1999, pp. 2272-2287, doi : 10.1016 / S0006-3495 (99) 77383-2 . PMID 10096922 . PMC 1300200 (free full text).
  16. ↑ Fluhler E, Burnham VG, Loew LM: Spectra, membrane binding, and potentiometric responses of new charge shift probes . In: Biochemistry . 24, No. 21, October 1985, pp. 5749-55. doi : 10.1021 / bi00342a010 . PMID 4084490 .