This service
includes sectioning,
ISNT labeling,
coverslipping
and labeling of
slides. As a result,
you will receive
up to 40 ISNT-labeled
sections per brain
or per tissue
block ready for
microscopic observations.
To express our
appreciation,
a set of Nissl
(or H & E)
stained sections
adjacent to those
used for ISNT
labeled sections
will also be provided
for you without
additional cost.
Procedure:
Unfixed frozen
tissue will be
cut on a cryostat
and mounted on
proteinase-resistant
microscope slides
(click to see
detail). Sections
cut through various
levels (or the
levels of your
choice) will be
processed on slides
for the detection
of neurodegeneration
with in situ nick
translation technique1.
This technique,
originally described
by Gold et al.
(1993)1, labels
the nuclei of
neurons undergoing
DNA fragmentation.
This method is
based on the ability
of DNA polymerase
I to catalyze
the polymerization
of deoxyuridines
onto the ends
of DNA fragments.
The integrated
biotins are amplified
and visualized
by the avidin-biotin-complex
methods2 (cf.
photo samples
below).
Remarks:
· A quotation
is required before
placing an order.
· The investigator
needs to provide
unfixed frozen
tissue.
· Please contact
us for more information.
References:
1. Gold R., Schmied
M., Rothe G.,
Zischler H., Breitschopf
H., Wekerle H.
and Lassmann H.
(1993) Detection
of DNA fragmentation
in apoptosis:
application of
in situ nick translation
to cell culture
systems and tissue
sections. J. Histochem.
Cytochem. 41:
1023-1030.
2. Hsu S. M.,
Raine L. and Fanger
H. (1981) Use
of avidin-biotin-peroxidase
complex (ABC)
in immunoperoxidase
techniques: a
comparison between
ABC and unlabeled
antibody (PAP)
procedures. J.
Histochem. Cytochem.
29: 577-580.
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