Peter S,

True Blue has been used extensively by neurochemists to label retrogradely
filled HRP-containing neurons.  In these experiments, ammonium molybdate is
used to stabilize the signal.  One commonly cited reference for this technique
is Olucha, J Neuroscience Methods 13, 131-138 (1985).  It works quite well.
Admittedly, I only tried the method once to develop an HRP-mediated
immunohistochemical signal on frozen sections.  In this experiment, the True
Blue substrate was at least 4-fold more sensitive than the DAB substrate.  I
next tried the technique on whole mount embryos, and that experiment was an
abysmal failure.  So I haven't pursued it further.  My results comparing True
Blue and DAB were very preliminary.  For instance, I hadn't titered my
antibody
out far enough to get a true picture of True Blue's sensitivity.  Nor had I
tried it with a variety of antibodies.  However, I've often wondered why it
isn't used more extensively in the histochemical community.  I wondered about
the long-term stability of the signal.  After three months, the signal is
still
robust.  Does anyone know why this substrate isn't used more extensively?

Karen D. Larison

Date:          Wed, 30 Dec 1998 08:21:19 +0000
From:          Peter =?iso-8859-1?Q?Siesj=F6?= <peter.siesjo@neurokir.lu.se>
Subject:       Trues Blue
To:            histonet@pathology.swmed.edu

Hello out there,
Has anyone successfully stained something with True Blue. In our seek for a
sensitive stain for cytokine detection in frozen sections we tried it. We
were not able to get a  stain for more than a couple of seconds, then it
disappeared despite different tricks recommended by the manufacturer. Or
should it be renamed False Blue?
Peter S

Peter Siesjo, M.D., Ph.D.
The Queen's Medical Center
Center for the Study of Neurological Disease
1356 Lusitana Street, U.T. 8th Floor
Honolulu, HI 96813, USA
Phone: (808) 537-7924
Phone: (home) (808) 924 1653
Fax:   (808) 537-7899
Email:peter.siesjo@neurokir.lu.se