Claudins are the most important components of the tight junctions at the interface of the basolateral and apical membranes of polarized epithelial and endothelial cells. They determine the barrier properties of cell–cell contact existing between two neighboring cells and regulate ...

Claudin-5 is a transmembrane tight junction protein highly expressed in brain endothelial cells, the site of the blood–brain barrier. The properties of the brain endothelial tight junction complex are considered to be dependent on claudin-5 cell–cell interaction, putting this prot ...

It is apparent that claudins are involved in signalling to and from cellular tight junctions (TJs) and control cell behaviour such as proliferation, differentiation, and migration. Methods to identify and measure specific claudins in TJs would, therefore, be useful to monitor TJ structu ...

The blood–brain barrier (BBB) has become a major focus of attention in cerebral pathophysiology and disease progression in the central nervous system. Endothelial tight junctions, the basal lamina, and perivascular astrocytes are jointly referred to as BBB or neurovascular unit. Aro ...

Claudins are transmembrane proteins that form the backbone of the tight junctions (TJs) at the blood–brain barrier (BBB). TJs are cellular structures that physically obstruct the inter-endothelial space and restrict the paracellular diffusion of blood-borne substances from the p ...

Metastatic spread of cancer is the major reason for patient mortality in human breast cancer. We examined expression of claudin-16 in human breast cells and tissues to identify a possible link between expression and aggressiveness in cells and between claudin-16 levels and patient progn ...

There are a number of disadvantages with conventional tissue immunohistochemistry for accurate �localisation of claudin proteins. Traditionally, tissue cryopreservation or formaldehyde fixation with wax embedding is utilised prior to sectioning and antibody localis ...

HIV-1 crosses the blood–brain barrier (BBB) early in the course of systemic infection and resides in brain macrophages and microglia. The integrity of the brain endothelium is regulated by intercellular tight junctions, which also play a critical role in HIV-1-entry into the brain. Disrupt ...

Investigation into the molecular mechanisms regulating normal renal physiology and pathophysiology has benefited from the development of microdissection techniques enabling sampling of specific cell populations or structures within the kidney. Laser-capture micro ...

Formalin-fixed and paraffin-embedded (FFPE) tissue is one of the most valuable resources available for molecular biological analysis on tissue after diagnostic histopathological examination. Gene expression profiles of FFPE can provide insights into molecular mechanis ...

We describe here a protocol optimized for formalin-fixed paraffin-embedded tissue sections that enables the detection of two antigens. This technique allows immunohistochemistry to be performed with detection systems allowing observation by light microscopy. This chapter ...

A number of techniques have been developed to use chemical, immunological, and molecular biology assays in histological material. Collectively termed histochemistry, these techniques have allowed us to better understand tissue and organ biology in situ. Success with each of these m ...

“A picture is worth a thousand words” goes the proverb. A poor picture however can be worse than saying nothing at all. This is particularly true for immunofluorescence pictures that in addition to their informative character bear an esthetic component. We here provide a panel of straightforwa ...

Apoptosis is an important process both in normal biology and in various pathologies and disease states. Apoptosis in tissue or cells can be detected in a number of ways. In tissue sections, electron microscopy can identify apoptosis by cellular and nuclear morphology, and in live cells, changes ...

In this chapter, we describe the use of immunohistochemical methods to detect hypoxia in tumour tissue sections, utilising antibodies specific for endogenous proteins hypoxia inducible factor 1 alpha (Hif1α) and glucose transporter 1 (Glut-1) and the exogenous compound pimonida ...

Magnetic resonance histology (MRH) has found considerable application in structural phenotyping in the mouse embryo. MRH employs the same fundamental principles as clinical MRI, albeit with spatial resolution up to six orders of magnitude higher than that in clinical studies. Criti ...

Extracellular matrix and cells are the components of connective tissue. Extracellular matrix provides tissue with tensile strength, elasticity, and resistance to compressive forces. Although immunohistochemistry is often used to study matrix constituents, a number of basic ...

This chapter deals with tissue preparation for subsequent detection of molecules in biological samples using immunocytochemistry and transmission electron microscopy. The aim of these methods is to localize specific molecules at high resolution in order to identify their subc ...

Immunohistochemistry has been used widely for the detection of proteins in brain tissue. The process can be performed on free-floating sections, but thicker sections are required than those required for processing on slides due to the “wear and tear” of the constant agitation that free-flo ...

We describe here a high-sensitivity in situ hybridization protocol, optimized for fresh-frozen brain sections, that enables the detection of two transcripts, at single cell resolution. Riboprobes directed against two mRNAs of interest are synthesized with nucleotides tagged wi ...