An optimized method for simultaneous demonstration of neurons and myelinated fiber tracts for delineation of individual trunco- and palliothalamic nuclei in the mammalian brain

Abstract

The truncothalamic complex has long been considered to be a nuclear group with "non-specific" projections. More recently, it is suggested that these thalamic nuclei play an important role in regulating distinct basal ganglia circuits. To further analyze the exact biological function of individual nuclei of the truncothalamic complex a simple and reliable technique for an exact delineation of distinct nuclei is desirable. Therefore, we evaluated and optimized several potential procedures for a combined visualization of neurons and myelinated fibers. Fiber staining with gold toning or immunocytochemical visualization of myelin basic protein shows high contrast and precision but precludes sufficient demonstration of neuronal cell bodies. When the most common technique for the simultaneous visualization of both structures, the Kluver-Barrera procedure, is used, demonstration of myelinated fibers is restricted when the technique is applied to cryostat or vibratome sections. In the present report this limitation was abolished. The final protocol includes lipid extraction prior to the incubation with Luxol Fast Blue and uses carefully characterized staining conditions for Luxol Fast Blue and cresyl violet rendering microscopically controlled differentiation steps unnecessary. The optimized Kluver-Barrera technique results in high precision localization of individual axons and cell bodies and thus permits an exact and simple delineation of individual nuclei in the vertebrate thalamus.

Therapeutic Benefit of Human Umbilical Cord Derived Mesenchymal Stromal Cells in Intracerebral Hemorrhage Rat: Implications of Anti-inflammation and Angiogenesis

“…Luxol fast blue/cresyl violet (Sigma) staining and the Fluoro-Jade B (FJB, Sigma) staining were performed according to published protocols [28,29]. Cells permeable to FJB were marked for cell death.…”

Role of Nrf2 in protection against intracerebral hemorrhage injury in mice

“…Luxol fast blue/Cresyl Violet, and Fluoro-Jade B (FJB) staining were performed according to published protocols [22,23]. Cells permeable to FJB are marked as degenerating neurons.…”

The development of an improved preclinical mouse model of intracerebral hemorrhage using double infusion of autologous whole blood

“…Mice (n = 6/group) were euthanized, and their brains were harvested, and rapidly frozen at −80°C. Frozen brain sections taken at 500 µm intervals by cryostat were stained with Luxol fast blue/Cresyl violet according to a published protocol (Geisler, et al, 2002). The areas of ipsilateral hemisphere and contralateral hemisphere were analyzed with advanced SPOT image software (Diagnostic Instruments Inc., Sterling Heights, MI).…”