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Fig. 1. Mice underwent injection of blood or saline into the prechiasmatic cistern
and then were euthanized and perfused 2 days later (
8
) . Photomicrographs of cortical
(
a
,
c
) and hippocampal (
b
,
d
) fi brinogen staining of microthromboemboli. After SAH, animals
have microthromboemboli in both cortical and hippocampal sections in comparison
to saline animals, where there is very little fi brinogen staining (from Sabri et al. (
8
) , with
permission).
applied to the assessment of immunohistochemical staining but
there can be a concern about variability due to technical factors
related to the staining (
9
).
4. Variations in the
Measurement
of Microthrom-
boembolism
We have assessed microthromboembolism in mice 2 days after
SAH. We have no experience with the assessment of microthrom-
boembolism at other time intervals.
4.1. When to Assess
Microthrombo-
embolism After SAH?
After animal death, the brains should be directly processed. The
method employed involves fi xation in 4% formaldehyde for 48 h
but shorter fi xation time or no fi xation after perfusion and use of
frozen sections could be used in order to avoid the need for anti-
gen retrieval. Regarding whether to perfuse the animal, if they
were not perfused then it would be diffi cult to differentiate fi brino-
gen left in situ from blood clotting postmortem. On the other
hand, perfusion could potentially fl ush out emboli from some
blood vessels. A method to avoid this could be to not perfuse and
4.2. Should Animals
Be Perfused?
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