Table 2.
Panel of stains useful in the interpretation of HS in epilepsy; those shown in bold are used in the 2013 ILAE classification of HS 9
| Histological preparation | Application | Methods/technical tips, limitation and pitfalls |
|---|---|---|
| Luxol fast blue/cresyl violet | Assessment of neuronal loss; these stains may be used interchangeably. NeuN is preferable for assessment of GCD. Neurofilament can highlight additional neuronal hypertrophy in CA4. Synaptophysin has also been used in the assessment of MFS, but it is not specific for mossy fibre pathway |
NeuN is fixation sensitive and gives less consistent staining in post-mortem tissues* Thicker sections (>10 μm ) also recommended for assessment of cyto-architecture with NeuN |
| NeuN | ||
| Synaptophysin | ||
| Map2 | ||
| Neurofilament | ||
| GFAP | Assessment of gliosis, patterns and distribution | Over-interpretation of endplate gliosis as sclerosis |
| GFAP-delta | ||
| CD34 | ||
| Timm method 16 | Assessment of mossy fibre re-organization | Timm stain: requires fixation of hippocampal slice from fresh specimen in buffered 1.2% sodium sulphide solution |
| Dynorphin ZnT3 |
Dynorphin: Thicker sections (>10 μm) recommended for better visualization of MFS | |
| Parvalbumin | Assessment of interneuronal groups. Antibody clones used in illustrations in current paper: calbindin D-28K (1:10 000, Swant, Switzerland ) Calretinin (polyclonal, 1:2000; Sigma, Saint Louis, MO, USA) NPY ( 1:4000, Sigma) Parvalbumin, (1:300 Swant, Switzerland) |
Parvalbumin can be fixation sensitive and give less consistent staining in post-mortem tissues* |
| Calbindin | ||
| Calretinin | ||
| Neuropeptide Y (NPY) |