Histochemical analysis

Muskhelishvili L, Thompson PA, Kusewitt DF, Wang C, Kadlubar FF. 2001. In situ hybridization and immuno-histochemical analysis of cytochrome P450 IBl expression in human normal tissues. J Histochem Cytochem 49 229-236. 

Figure 21.2 Histochemical analysis of / -galactosidase gene expression in liver. Mice were injected with 1.6ml saline containing various amounts of pCMV-LacZ plasmid DNA. Animals were sacrificed eight hours post injection and liver sections were made using cryostat. Sections (A, B, C and D) were stained with X-gal solution followed by eosin for counter-stain. Sections (E, F, G and H) were stained by a standard hematoxylin/eosin staining method. Sections were made from animals each receiving 0 (A, E), 0.5 (B, F), 2.5 (C, G) and 25 pg (D, H) of pCMV-LacZ. (25x). (see Color Plate 14)...

Color Plate 14 Histochemical analysis of P -galactosidase gene expression in liver, 424... 

Histochemical analysis of /3-galactosidasc gene expression in liver 424... 

Forerunners of nonisotopic immunoassay had already appeared before radioimmunoassay was developed. For example, nephelometry is based on precipitation, which is known as the classical immune reaction, and the ideas of particle immunoassay and viroimmunoassay seem to have developed from the hemagglutination test. The principles of enzyme and fluorescence immunoassay had already been used as enzyme and fluorescence antibody techniques in histochemical analysis. In 1971, two groups reported use of an enzyme immunoassay (E5, V2). Leute et al. reported spin immunoassay, which has spurred recent development of nonisotopic immunoassays (L5). 

Gerfen CR, McGinty JF, Young WS, 3d. (1991) Dopamine differentially regulates dynorphin, substance P, and enkephalin expression in striatal neurons in situ hybridization histochemical analysis. J Neurosci 77 1016-1031. 

The antiatherosclerotic effect of proanthocyanidin-rich grape seed extracts was examined in cholesterol-fed rabbits. The proanthocyanidin-rich extracts [0.1% and 1% in diets (w/w)] did not change the serum lipid profile, but reduced the level of the cholesteryl ester hydroperoxides (ChE-OOH) induced by 2,2/-azo-bis(2-amidinopropane-dihydrochloride (AAPH), the aortic malonaldehyde (MDA) content and severe atherosclerosis. The immuno-histochemical analysis revealed a decrease in the number of the oxidized LDL-positive macrophage-derived foam cells on the atherosclerotic lesions of the aorta in the rabbits fed the proanthocyanidin-rich extract. When the proanthocyanidin-rich extract was administered orally to the rats, proantho-cyanidin was detected in the plasma. In an in vitro experiment using human plasma, the addition of the proanthocyanidin-rich extract to the plasma inhibited the oxidation of cholesteryl linoleate in the LDL, but not in the LDL isolated after the plasma and the extract were incubated in advance. From these results, proanthocyanidins of the major polyphenols in red wine might trap ROSs in the plasma and interstitial fluid of the arterial wall, and consequently display antiatherosclerotic activity by inhibiting the oxidation of the LDL [92]. 

Sergeeva, L.I., Vonk, J., Keurentjes, J.J.B., van der Plas, L.H.W., Koomneef, M., and Vreugdenhil, D., 2004, Histochemical analysis reveals organ-specific quantitative trait loci for enzyme activities in Arabidopsis. Plant Physiol. 134 237-245. 

Coincident with this new technique for procurement of human bone biopsies was the development of quantitive methods of bone analysis.12 These methods include histochemical analysis of both decalcified and unde-calcified42 48 bone sections, microradiography,44 tetracycline labeling45 and autoradiography.42 The latter two techniques require administration of a tetracycline antibiotic or isotopic tracer prior to procurement of the biopsy. Undecalcified thin sections, prepared with the use of a Jung microtome after the bone core is fixed, dehydrated and embedded in methacrylate,45 are analyzed by intersect and point count methods46 47 which permit three-dimensional assessment.48 49 Tetracycline antibiotics deposit in vivo in sites of bone formation constituting markers which can be studied in undecalcified sections by fluorescence microscopy.45 47 This represents the safest and best tissue time marker for microscopic measurement of bone formation dynamics. 

Q.29.1 We recognize that each level of exploration carries a cost vs. benefit problem. Perhaps this is best called the forest or trees conflict. With greater visualization of detail there is a loss of knowledge of the overall organization of the system. This is a common problem in microscopy. What are the problems to be considered in histochemical analysis or the use of fluorescent probes. What is gained and what is lost with each of these observational levels. Is one level better than any other ... 

Histochemical analysis stem from the use of chemical reactions with a... 

Histochemical analysis of enzymes in rat adrenal sections demonstrated that WADPH-dichlorophenolindophenol reductase of the zona fasciculata-ret-icularis (z.f-r.) is stimulated by ACTH whereas HADPH or MDH quinone reductase, NADPH and NADH oxidases or NADPH-NAD transhydrogenases are not85. 

For all genetically modified mice, it is necessary to test first whether homozygous mutant mice are born at Mendelian ratio (embryonic lethality ), are viable, grow normally, are fertile, and have a normal life span. A histochemical analysis should then be carried out of all tissues that express the mutated gene. More specific analyses (e.g., immunofluorescence, electron microscopy or biochemical analysis) of tissues or primary cells derived from the mutant mice depends on the phenotype. 

Inagaki S, Shiosaka S, Takatsuki K, lida H, Sakanaka M, Senba , Hara Y, Matsuzaki T, Kawai Y, Tohyama M (1982) Ontogeny of somatostatin-containing neuron system of the rat cerebellum including its fiber connections an experimental and immuno-histochemical analysis. Dev Brain Res 3 509-527... 

After 8 weeks, some of the recipient mice were sacrificed and histochemical analysis was performed on tissue sections of... 

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