![]() Pistorio AL, Hendry SH, Wang X (2006) A modified technique for high-resolution staining of myelin. Pinaud R, Mello CV, Velho TA, Wynne RD, Tremere LA (2008) Detection of two mRNA species at single-cell resolution by double-fluorescence in situ hybridization. Pinaud R, Jeong JK (2010) Duplex in situ hybridization in the study of gene co-regulation in the vertebrate brain. Luo P, Haines A, Dessem D (2001) Elucidation of neuronal circuitry: protocol(s) combining intracellular labeling, neuroanatomical tracing and immunocytochemical methodologies. Li L, Yun SH, Keblesh J, Trommer BL, Xiong H, Radulovic J, Tourtellotte WG (2007) Egr3, a synaptic activity regulated transcription factor that is essential for learning and memory. Li JL, Wang D, Kaneko T, Shigemoto R, Nomura S, Mizuno N (2000) The relationship between neurokinin-1 receptor and substance P in the medullary dorsal horn: a light and electron microscopic immunohistochemical study in the rat. Kluver H, Barrera E (1953) A method for the combined staining of cells and fibers in the nervous system. Hasegawa R, Takami S, Nishiyama F (2008) Immunoelectron microscopic analysis of the distribution of tyrosine kinase receptor B in olfactory axons. Gage GJ, Kipke DR, Shain W (2012) Whole animal perfusion fixation for rodents. Elsevier, Amsterdam, pp 50–121Ĭarriel V, Garzon I, Alaminos M, Campos A (2011) Evaluation of myelin sheath and collagen reorganization pattern in a model of peripheral nerve regeneration using an integrated histochemical approach. In: Bjoklund A, Hokfelt T (eds) Handbook of neuroanatomy. Keywordsījorklund A (1983) Fluorescence histochemistry of biogenic monoamines. Due to demyelination being a highly researched area, this method is very applicable in current day studies of demyelinating diseases and animal models thereof. (9) Finally, we introduce a myelin sheath stain, Luxol fast blue, with practicable protocol. (8) The methodology of hematoxylin and eosin (HE) stain, a broadly used histology stain method in both research and clinical labs, will be discussed here with detailed protocol. Two Nissl stains detailed here are cresyl violet and neutral red stains. (7) Protocols for Nissl staining, another commonly used neuron stain, and relevant photos. This section also highlights examples of new applications of this long-established method with Golgi’s stain kit in modern neuroscience research with representative images. (6) A protocol for a well-known traditional neural stain, Golgi’s stain, is presented here, with representative microimages. We will also illustrate a protocol for production of self-made siliconized slides used for plate embedding. To overcome the obstacle of extracting such a small area while ensuring it contains the desired labeling, a plate embedding method is used, which is described in this chapter. These ultrathin sections are generally 1.5 mm long and 0.5 mm wide. (5) Following histochemical staining, the stained structures need to be further observed under EM. We described detailed protocols for tissue treatment using freeze and thaw and ABC kit to reveal immunostain, two key steps in all immuno-EM. In this chapter, procedures introduced will be based on cutting 50 μm sections for EM histochemical stains and plate embedding. (4) Vibratome sectioning and application in histochemical staining for electron microscopy (EM) studies. (3) Paraffin embedding and sectioning, in which a general procedure is introduced. Examples will be based on the author’s own experiences. (2) Cryoprotection and frozen section cutting, with emphasis on the importance of cryoprotection for obtaining superior histological staining. ![]() The selectively introduced methods in this chapter are as follows: (1) Transcardial perfusion, especially on rats and mice, with a supplemental video referenced online to show perfusion procedure for adult rats. History, development, and novel application of those methods in neuroscience will be presented herein so students can fully grasp the topic of brain tissue preparation, sectioning, and staining. This chapter will summarize commonly used methods of brain tissue preparation, sectioning, and staining.
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