From gauthier@biomail.ucsd.edu Thu May 29 22:30:26 2003
Date: Thu, 29 May 2003 22:30:11 -0700 (PDT)
From: Jeffrey Gauthier <gauthier@biomail.ucsd.edu>
To: Jeffrey Gauthier <gauthier@biomail.ucsd.edu>
Subject: Calcium Dyes

Two strategies:

1)  In rats, cause the ultraviolet cones to adapt (using a
frequency outside the dye's excitation frequency so that the dye doesn't
bleach).

2)  In guinea pigs, use only that half of the retina which has M cones.


Comparison:
Doc Ophthalmol 2003 May;106(3):243-9
The ERG of guinea pig (Cavis porcellus): comparison with I-type monkey and
E-type rat.


Guineau pig eyes:

Neuron 1994 Nov;13(5):1159-66 
Two different visual pigments in one retinal cone cell.

The retina of the mouse, rabbit, and guinea pig is divided into a superior
area dominated by green-sensitive (M) cones and an inferior area in which
cones possess practically only short wavelength-sensitive
(S) photopigments. The present study shows that the transitional zone
between these retinal areas is populated by cones labeled by both the M
and S cone photopigment-specific antibodies


Invest Ophthalmol Vis Sci 2002 May;43(5):1662-5
Visual pigment coexpression in Guinea pig cones: a microspectrophotometric
study.


Why rats are a problem:

Nature 1991 Oct 17;353(6345):655-6
Retinal receptors in rodents maximally sensitive to ultraviolet light.

J Exp Biol 2001 Jul;204(Pt 14):2439-46
Cone-based vision of rats for ultraviolet and visible lights.

Invest Ophthalmol Vis Sci 1995 Apr;36(5):851-63
Ultraviolet and green light cause different types of damage in rat retina.



Fura-2-AM
Excitation = 355 nm
Emission = 495 nm
Sigma Product Number: 47989



Alternative Dyes:


Quin-2
Excitation = 332 nm
Emission = 493 nm 
Sigma Product Number: 08520
R.Y. Tsien, Biochemistry 19, 2396 (1980)
R.Y. Tsien, et al., J. Cell Biol. 94, 325 (1982)


Thio-H-AM
Excitation = 322 nm
Emission = 500
Sigma Product Number: 29218
Soluble in DMSO
Nonpolar AM-esters passively cross the plasma membrane and accumulate in
the cytoplasm of living cells. After cleavage of the protecting groups by
intracellular esterases, the polar fluorophore remains trapped within the
cell.

Also Thio-H (without AM)


Indo 1
Excitation = 330 nm
Emission = 398
Sigma Product Number: 57180
unusually large shift in fluorescence emission from 480 nm to 400 nm
G. Grynkiewicz, et al., J. Biol. Chem. 260, 3440 (1985)
K.G. Lazzari, et al., J. Biol. Chem. 261, 9710 (1986)
M.K. Bijsterbosch, et al., Biochem. Biophys. Res. Comm. 137, 500 (1986)

(also Indo 1-AM)


Possibly more at

http://www.sigmaaldrich.com/Brands/Fluka___Riedel_Home/Analytical/Fluorescent_Probes/Chelators_and_Ion_Probes/Calcium_Indicators.html