Which Voltage Sensitive Dye?
- 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.
|
|
- Rat neocortex slice (10/99)
Jian Young Wu (wuj@giccs.georgetown.edu)
| Signal type: |
Absorption |
| Dye name: |
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 et al.(1999), Propagation activation
during oscillation. J.Neuroscience 19:5005-5015
- Rat & mouse neocortex and
olfactory bulb slices (10/99)
-
Asaf Keller (akeller@umaryland.edu)
| Signal type: |
Absorption |
| Dye name: |
RH-155 |
| Supplier: |
Molecular Probes |
| Concentration: |
100 ćM |
| Staining duration: |
20 to 60 min. |
| Fractional change
(F/F): |
up to 2x10-2 |
| Pharmacology: |
negligible |
| Phototoxicity: |
negligible |
- Guinea pig and mouse submucosal
and myenteric neurones.*(6/99)
-
Michel Neunlist ( mneunlis@win.tiho-hannover.de )
-
Michael Schemann ( mschem@win.tiho-hannover.de )
| Signal type: |
Fluorescence |
| Dye name: |
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: |
| 10 minutes (whole preparation) |
| 1-2 min (local application) |
|
| 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, Peters, and Schemann: Multisite optical recording
of excitability in the enteric nervous system. Neurogastroenterol.
Mot. 1999. (in press)
- Rat, hippocampus, piriform
cortex, and spinal cord (6/99)
-
Meyer Jackson* ( mjackson@macc.wisc.edu )
| Signal type: |
Fluorescence |
| Dye name: |
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 |
- Rat, neonatal cardiac myocytes
in culture* (6/99)
-
Brian Salzberg ( bmsalzbe@mail.med.upenn.edu )
*Rohr and Salzberg (1994) Biophysical Journal 67:1301-1305
["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."
- Guinea-pig, submucous plexus
neurons in situ* (6/99)
-
Brian Salzberg ( bmsalzbe@mail.med.upenn.edu )
Ana Lia Obaid (
obaid@mail.med.upenn.edu )
| Signal type: |
Fluorescence |
| Dye name: |
di-8-ANEPPS |
| Supplier: |
Molecular Probes |
| Catalogue number: |
D-3167 |
| Concentration: |
100 ug/ml |
| Staining duration: |
> 5 minutes (dye left in bath) |
| 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 et al. (1999) J. Neuroscience 19: 3073-3093.
- Rat and guinea pig, neocortex
and hippocampus Human, neocortex obtained during epilepsy
and tumor surgery (6/99)
-
Erwin Speckmann ( speckma@uni-muenster.de )
| Signal type: |
Fluorescence |
| Dye name: |
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 |
- 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. J Neurosci 15: 1392-1405
- Embryonic chick and rat heart§ Adult frog and rat heart
Embryonic chick and rat CNS* (6/99)
-
Kohtaro Kamino ( kamino.phy2@med.tmd.ac.jp )
| 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.
* 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.
- Aplysia abdominal ganglion. (6/99)
-
Larry Cohen ( lawrence.cohen@yale.edu )
| Signal type: |
Absorption |
| Dye names: |
RH155 (NK3041) and RH482 (NK3630) |
| Supplier: |
Nippon Kankoh Shikiso Kenkyu-sho |
| Concentration: |
0.1-0.2 mg/ml |
| Staining duration: |
10 minutes |
| Fractional change
(F/F): |
10-4 to 10-3 |
| Pharmacology: |
needs watching |
| Phototoxicity: |
small |
- Turtle olfactory bulb. (6/99)
-
Larry Cohen ( lawrence.cohen@yale.edu )
| Signal type: |
Fluorescence |
| Dye name: |
RH414 |
| Supplier: |
Molecular Probes |
| Catalogue number: |
T-1111 |
| Concentration: |
0.05-0.2 mg/ml |
| Staining duration: |
60 minutes |
| 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. J.
Neurosci. 20(2):749-762.
