Nervous system

Roulois, Aude

doi:10.1002/9781118789568.ch8

Cited by 3 publications

(1 citation statement)

References 57 publications

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Central nervous system (CNS) tissues are very sensitive to handling, fixation, and processing artifacts which can affect downstream histopathologic analysis. 1,2,7,9,12,16 These artifacts include the presence of contracted, intensely basophilic neurons (known as dark neuron artifact); artifactual neuronal and white matter vacuolation; clear halos surrounding oligodendrocytes; perivascular cleft formation; and retraction spaces. It is important to reduce artifacts in diagnostic and experimental studies because they may be misinterpreted as antemortem lesions 11 and they may obscure or complicate the diagnosis of other lesions (eg, if artifactual vacuolation is present, the pathologist may not appreciate subtle vacuolation occurring secondary to neurological disease).

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confidence: 99%

Perfusion with 10% neutral-buffered formalin is equivalent to 4% paraformaldehyde for histopathology and immunohistochemistry in a mouse model of experimental autoimmune encephalomyelitis

et al. 2022

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Intravascular (IV) perfusion of tissue fixative is commonly used in the field of neuroscience as the central nervous system tissues are exquisitely sensitive to handling and fixation artifacts which can affect downstream microscopic analysis. Both 10% neutral-buffered formalin (NBF) and 4% paraformaldehyde (PFA) are used, although IV perfusion with PFA is most commonly referenced. The study objective was to compare the severity of handling and fixation artifacts, semiquantitative scores of inflammatory and neurodegenerative changes, and quantitative immunohistochemistry following terminal IV perfusion of mice with either 10% NBF or 4% PFA in a model of experimental autoimmune encephalitis (EAE). The study included 24 mice; 12 were control animals not immunized and an additional 12 were immunized with PLP139–151 subcutaneously, harvested at day 20, and fixed in the same fashion. Equal numbers (4 per group) were perfused with 10% NBF or 4% PFA, and 4 were immersion-fixed in 10% NBF. NBF-perfused mice had less severe dark neuron artifact than PFA-perfused mice ( P < .001). Immersion-fixed animals had significantly higher scores for oligodendrocyte halos, dark neuron artifact, and perivascular clefts than perfusion-fixed animals. Histopathology scores in EAE mice for inflammation, demyelination, and necrosis did not differ among fixation methods. Also, no significant differences in quantitative immunohistochemistry for CD3 and Iba-1 were observed in immunized animals regardless of the method of fixation. These findings indicate that IV perfusion of mice with 10% NBF and 4% PFA are similar and adequate fixation techniques in this model.

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confidence: 99%