Vibratome

Evers P, Uylings HB. Effects of microwave pretreatment on immunocytochemical staining of vibratome sections and tissue blocks of human cerebral cortex stored in formaldehyde fixative for long periods. /. Neurosci. Methods 1994 55 163-172. 

Preembedding hybridization where permeabilized cells or vibratome sections are hybridized, and then postfixed and embedded as described in Section 3.1.2. 

The jar is allowed to cool for 15 min, and then the tissue blocks are placed in TBS (pH 7.6). Thick sections (95 xm) are cut on a vibratome, collected in plastic vials containing TBS, washed several times in TBS for 1 hr, and immunostained according to standard procedures given in this volume. These thick sections allow staining of whole neurons, including neuronal processes, to distinguish different morphological types. 

ANTIGEN RETRIEVAL IN NEURONAL TISSUE SLICES BEFORE VIBRATOME SECTIONING... 

Fig. 13.2. Preferred association of LCM with tumor cells in vivo. Vibratome sections of brain of rats bearing 9L tumor that have been injected i.v. with diO-LCM, and sacrificed 2 min later. Sections in panels A and C were stained with TR-conjugated WGA. Specimens in panels B and C were observed by fluorescence microscopy using fluorescein optics. The scale bar represents 100 pm. (Taken from ref. 531.)...

Fig. 13.3. This figure demonstrates the distribution of fluorescently tagged LCM in brain parenchyma analyzed by confocal laser scanning microscopy. Rats bearing 9L tumors were administered diO-LCM and sacrificed 2 minutes later. Vibratome sections were counterstained with TR-WGA which binds to tumor cells and distinguishes the tumor area from the surrounding normal tissue. Comparison of the area stained with TR-WGA (tumor cells) (B) and that stained with diO (A) indicates that LCM were associated with a large portion of the tumor. (Taken from ref. 531.)...

Cutting thick, frozen sections using a cryostat (20—40 xm thick sections) or using a vibratome to cut 50 jrm sections of unfrozen tissue (remove the tendons to facilitate sectioning) gives good results. Muscle fibers can also be teased directly on slides to obtain individual fibers. 

Male Wistar rats (280—320 g of weight) were deeply anaesthetized with halo-thane and subsequendy decapitated. Brain slices were prepared according to a previously described procedure (Geracitano et al., 2003). Briefly, the brain was rapidly removed and corticostriatal coronal slices (300 /nn) were cut from a tissue block with the use of a vibratome (at 20-25 °C) in artificial cerebrospinal fluid solution (ACSF). 

Fixed tissue fragments are usually chopped with a semi-automatic tissue sectioning system (e.g., the vibratome) or with a razor blade into sections of about 20-50 pm. The sections are stained and washed... 

Fig. 59. 5 -Nucleotidase immunohistochemical staining of rat cerebellum. A. Immunofluorescence. B. PAP-method. Enzyme activity is predominantly found within the molecular layer on Bergmann glial fibers (long arrows). Purkinje cells are surrounded by fine rims of reaction product (small arrows). Within the granular layer 5 -nucleotidase activity is diffusely scattered between granule cells (arrow heads). Vibratome sections. C. Longitudinally sectioned Bergmann glia cell processes (B) of the molecular layer of rat cerebellum. Fine DAB reaction product is located on adjacent membranes of these processes (arrows). Bars in A,B = 50 /tm, in C = 0.5 nm. Schoen et al. (1987).

Floating section immunocytochemistry - performed on free-floating, thick section (25-100 mm) cut on a Vibratome or freezing microtome with antibodies penetrating deeper because of the greater movement of the section floating both sides of the section will be exposed to the antibodies. 

Vibratome - a device used on delicate, fixed tissue before sectioning a vibrating razor blade cuts large pieces of tissue into sections 25-500 p,m for freezing or processing. 

Ma, B., Jablonska, J., Lindenmaier, W., and Dittmar, K. E. J. (2006) Immunohistochemi-cal study of the reticular and vascular network of mouse lymph node using vibratome sections. Acta Histochem. 109, 15-28. 

Fig. 8.6 Immunohistochemical localization of the neuropeptide SIFamide in the brain of the giant robber crab Birgus latro (Harzsch and Hansson, unpublished data black-white inverted fluorescent images), (a) Horizontal vibratome section of the median brain. The olfactory lobes (OLs) and the hemiellipsoid bodies (HE) dominate the brain, (b) Detail of the hemiellipsoid body showing its layered structure, (c) The OL seems to be composed of three units...

Vibratome (Vibratome Series 2(XX), Technical Products International Inc., St. 

Incubation of Vibratome Sections in Secondary Antibody, Immunoperoxidase Deveeopment, and Initial Screening by Light Microscopy... 

After twice-daily treatment for 14 d, animals were deeply anesthetized with 300 mg/kg of chloral hydrate (ip). They were perfused through the left ventricle, initially with a cold saline solution (first washing solution) and subsequently with a fixative solution. Brains were removed and kept in the same cold fixative solution for 4 h. Then, brains were washed three times in the second washing solution and left in this washing solution for 18 h at 4°C. Coronal and saggital 40-pm-thick brain sections were obtained using a vibratome. The sections were cryoprotected by immersing them in a cryoprotective solution and stored at -20°C until the immunohistochemical studies were performed. 

Preparation of brain slices is an indispensable procedure for a variety of experiments. The goal of the slice preparation is to obtain a thin piece of brain tissue containing the cells of interest and to maintain the slice in a viable (although artificial) condition that is similar to its in vivo environment. In this chapter we describe procedures that are fundamental for brain slice preparation. Because the hippocampus is the most widely used tissue for brain slices, Its isolation will be used here to illustrate the steps of the procedure. Two slicing methods, using either a vibratome or a tissue chopper, are described. Our discussion of these methods covers the steps from the decapitation of the animal to the storage of slices in artificial cerebrospinal fluid (aCSF). Some of the... 

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