[Protocol for the combination of neurohistological techniques on vibratome obtained sections].
BACKGROUND
The histological study of the nervous system requires the use of special techniques. Currently, no methods are available to visualize simultaneously all the cellular constituents of nervous tissue.


OBJECTIVE
A protocol was adapted for staining nervous tissue by modification of a formerly difficult procedure.


METHODS
Slices of brain and spinal cord, 4 mm thick, were taken from adult mice, previously fixed by intracardiac perfusion with 4% paraformaldehyde. Vibratome sections were obtained with thickness of 15-25 µm. These were mounted on glass slides prepared with gelatin as an adhesive. The preparations were subjected to staining protocol Luxol Fast Blue-PAS-hematoxylin (LPH) combined with silver staining method (LPH-Holmes).


RESULTS
LPH technique yielded an excellent differentiation of gray and white matter in all regions of the nervous system. A panoramic view of the gray matter was colored pink in contrast to the myelinated nerve fibers and tracts which were light blue. The combination LPH-Holmes retained the staining characteristics but significantly improved the demarcation of axons and tracts.


CONCLUSIONS
A protocol was standardized for the LPH and LPH-Holmes nervous tissue stains applied in combination to tissue slices obtained with a vibratome. The method was shorter, less wasteful and less expensive than the original and also better preserved the integrity of nervous tissue.
Lukas
L
Tamayo-Orrego
Orlando
O
Torres-Fernández
D000818:Animals; D001921:Brain / Q000648:ultrastructure*; D006416:Hematoxylin; D007211:Indoles; D051379:Mice; D008867:Microtomy / Q000379:methods*; D009186:Myelin Sheath / Q000648:ultrastructure; D016612:Paraffin Embedding; D010503:Periodic Acid-Schiff Reaction; D016622:Silver Staining; D013048:Specimen Handling; D013116:Spinal Cord / Q000648:ultrastructure*; D013194:Staining and Labeling / Q000379:methods* / Q000592:standards
D007211:Indoles; C018588:Luxol Fast Blue MBS; D006416:Hematoxylin