|
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.
|
