Laminin immunohistochemistry in brain is dependent on method of tissue fixation

Normal adult and lesioned rat and mouse brains were fixed by formaldehyde perfusion by two methods that differ primarily in the length of the post-fixation period. Sections were subsequently immunostained using monoclonal and polyclonal antibodies to laminin. With relatively short post-fixation periods (up to 4 h), vascular basement membrane (BM)-laminin was immunostained, but intraneuronal laminin-like immunoreactivity was faint. With longer post-fixation periods (18-24 h), intraneuronal laminin-like immunoreactivity was distinct, while vascular BM-laminin immunoreactivity was reduced drastically. These findings are particularly relevant to studies examining laminin immunoreactive blood vessels in response to lesions, especially ischemic stroke. In fact, the present results suggest that the apparent neovascularization or up-regulation of vascular BM-laminin following CNS injury likely relates to differences in regional tissue fixation.