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What follows is an informal compilation of the voltage-sensitive dyes that have been used in a variety of preparations. The information includes dye source, signal size, staining conditions, and pharmacological- and photo-toxicity. In a few instances several dyes have been tested. In some cases the data is presented directly, in others a reference is given, and in some cases both. You can contact the individual scientist directly for additional or more up-to-date information.

The dyes with the label WW were synthesized by Jeff Wang and Alan Waggoner, then at Amherst College. Dyes labeled RH were synthesized by Rina Hildesheim and Amiram Grinvald at the Weizmann Institute. Dyes labeled JPW were synthesized by Joe Wuskell and Les Loew at the University of Connecticut Health Center. We depend on them.

Dissolving the dyes can be tricky in that they may look dissolved but still be in small crystals. You can check this out by filtering through a Millipore filter. If the dye was not dissolved, try modest warming (e.g. 50°C). Hydrophobic dyes will require more extensive procedures involving ethanol and Pluronic F127.

Aplysia abdominal ganglion (6/99)
Clione and Helix ganglia. (9/01)
Embryonic chick and rat heart Adult frog and rat heart Embryonic chick and rat CNS (6/99)
Guinea pig and mouse submucosal and myenteric neurones.(6/99)
Guinea-pig, submucous plexus neurons in situ* (6/99)
Helix neurons, rat cortical neurons; internally injected dyes (6/99)
Rat and guinea pig, neocortex and hippocampus Human, neocortex obtained during epilepsy and tumor surgery (6/99)
Rat & mouse neocortex and olfactory bulb slices
Rat, hippocampus, piriform cortex, and spinal cord. (6/99)
Rat neocortex slice
Rat, neonatal cardiac myocytes in culture (6/99)
Turtle olfactory bulb (6/99)

 
 
Signal type: Absorption
Dye names: RH 479 (JPW 1131) RH482 (JPW 1132, NK3630)
Supplier: JPW dyes from L. Loew at U. Conn; NK dyes from Nippon Kankoh
Staining duration: 0.05-0.1 mg/ml for 30 min
Fractional change ( F/F): 0.1% to 0.5%
Pharmacology: not detectable
Phototoxicity: not significant @ exposure < 120 second
 



Signal type: Absorption
Dye names: RH479
Supplier: Amiram Grinvald, Les Loew
Concentration: 0.3-0.4 mg/ml with pluronic and a very low concentration of DMSO
Staining duration: 2 times 5 minutes with wash in between, done at low temperature (7 °C)
Fractional change ( F/F): 10-4 to 10-3
Pharmacology: small
Phototoxicity: small
Bleaching: 2 minutes
 



Signal type: Absorption
Dye names: NK2761, NK2776, NK3224, NK3225
Supplier: Nippon Kankoh Shikiso Kenkyu-sho
Concentration: 0.2 mg/ml
Staining duration: 20 minutes
Signal-to-noise ratio: 5:1 (guinea pig); 3:1 (mouse)
Fractional change ( F/F): 10-4 to 10-3
Pharmacology: negligible
Phototoxicity: negligible

#Kamino, K., Hirota, A., and Fujii, S. (1981 ) Localization of pacemaker activity in early embryonic heart monitored using voltage sensitive dyes. Nature. Apr 16;290(5807):595-7
*Momose-Sato, Y., Sato, K., Sakai, T., Hirota, A., Matsutani K., and Kamino, K. (1995) Evaluation of optimal voltage sensitive dyes for optical monitoring of embryonic neural activity. J. Membrane Biology 144: 167-176.




Signal type: Fluorescence
Dye names: Di-8-ANEPPS
Supplier: Molecular Probes
Catalogue number: D-3167
Concentration:
staining whole preparation:20µM (stock: 10.3 mM; 75% DMSO and 25% Pluronic F-127)
local staining with pipette:200µM
Staining duration:
whole preparation:10 minutes
local application:1-2 min
Signal-to-noise ratio: 5:1 (guinea pig); 3:1 (mouse)
Fractional change ( F/F): 0.1 - 0.5%
Pharmacology:
whole preparation:not tested
local application:negligible
Phototoxicity: none after 40 seconds of recording

*Neunlist M., Peters S. and Schemann M. (1999) Multisite optical recording of excitability in the enteric nervous system. Neurogastroenterol Motil  Oct;11(5):393-402



Signal type: Fluorescence
Dye names: Di-8-ANEPPS
Supplier: Molecular Probes
Catalogue number: D-3167
Concentration: 100 ug/ml
Staining duration: 10 minutes
Relative signal size: only one tested
Fractional change ( F/F):  
Pharmacology: none
Phototoxicity: significant phototoxicity (limited by restricting O2 with glucose oxidase and catalase)

*Obaid AL, Koyano T, Lindstrom J, Sakai T, Salzberg BM. (1999) Spatiotemporal patterns of activity in an intact mammalian network with single-cell resolution: optical studies of nicotinic activity in an enteric plexus. J Neurosci. Apr 15;19(8):3073-93.



Helix neurons*, rat cortical neurons; internally injected dyes (6/99)
Dejan Zecevic · dejan.zecevic@yale.edu
Absorption dyes
  30 dyes were tested (twenty pyrazolone-oxonols molecules, five merocyanine dyes, three barbituric-acid oxonol dyes, and 2 styryl dyes; see legend to Fig.3). The best results were obtained with two positively charged pyrazolone-oxonol dyes (JPW1177 and JPW1245), and two negatively charged merocyanine dyes (WW375 and JPW1124). The relative signal size for the best four: 1.
Signal type:Fluorescence
Dye name:RH461
Relative signal size:0.5
Dye name:RH437
Relative signal size:0.5
Dye name:JPW1063
Relative signal size:1
 
Signal type:Fluorescence
Dye name:JPW1114
Relative signal size:30
Supplier:Molecular Probes
Catalogue number:D-6923
Concentration:3 mg/ml in electrode
Staining duration:60 minutes
Pharmacology:none if careful
Phototoxicity:small in Helix, larger in cortical neurons
Dye name:JPW3028
Relative signal size:35

*Antic S, Zecevic D (1995) Optical signals from neurons with internally applied voltage-sensitive dyes. Journal of Neuroscience Feb;15(2):1392-405.



Signal type: Fluorescence
Dye names: RH795
Supplier: Mo Bi Tec, Wagenstieg 5, 37077 Göttingen
Catalogue number: R-649
Concentration: 12.5 ug/ml
Staining duration: 60 minutes
Signal-to-noise ratio: 60 min (humans), 120 minutes (animals)
Fractional change ( F/F):  
Pharmacology:  
Phototoxicity: negligible





Signal type: Absorption
Dye names: RH-155
Supplier: Molecular Probes
Concentration: 100 æM
Staining duration: 20 to 60 min.
Fractional change ( F/F): up to 2x10-2
Pharmacology: negligible
Phototoxicity: negligible



 
Signal type: Fluorescence
Dye names: RH414
Supplier: Molecular Probes
Catalogue number: T-1111
Concentration: Usually 200 uM, occasionally 50 uM
Staining duration: 10 minutes to an hour
Relative signal size: only one tested
Fractional change ( F/F): 0.1 - 0.5%
Pharmacology: none
Phototoxicity: severe unless 1 mM sulfate is present





Signal type: Absorption
Dye names: RH 479 (JPW 1131) RH482 (JPW 1132, NK3630)
Supplier: JPW dyes from L. Loew at U. Conn; NK dyes from Nippon Kankoh
Staining duration: 0.05-0.1 mg/ml for 30 min
Fractional change ( F/F): 0.1% to 0.5%
Pharmacology: not detectable
Phototoxicity: not significant @ exposure < 120 second

* Wu JY, Guan L, Tsau Y. (1999), Propagation activation during oscillation. J.Neuroscience  Jun 15;19(12):5005-15.



* Rohr S, Salzberg BM. (1994) Multiple Site Optical Recording of Transmembrane Voltage (MSORTV) in Patterned Growth Heart Cell Cultures: Assessing Electrical Behavior, with Microsecond Resolution, on a Cellular and Subcellular Scale. Biophysical Journal 67(3):1301-15.



Signal type: Fluorescence
Dye names: RH414
Supplier: Molecular Probes
Catalogue number: T-1111
Concentration: 0.05-0.2 mg/ml
Staining duration: 60 minutes
Relative signal size:  
Fractional change ( F/F): 10-3 to 10-2
Pharmacology: Pharmacology not detected
Phototoxicity: small

For results with additional dyes on the turtle see:
Ying-wan Lam, Lawrence B. Cohen, Matt Wachowiak, and Michal R. Zochowski, (2000), Odors elicit three different oscillations in the turtle olfactory bulb. Journal of Neuroscience,  Jan 15;20(2):749-62.



 
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