Cytochemical procedures

Because disrupted tissue preparations were unsatisfactory, attempts were made to work either with more organized systems such as tissue slices (liver-Krebs) or to identify and isolate the intracellular organelles involved in the reactions. Cytochemical procedures were developed in the 1930s and 1940s to locate sites of reaction in situ in cells (Chapter 9). Examination of cell ultrastructure became possible when the electron microscope was introduced after 1945. Techniques for the isolation of cell organelles, notably mitochondria, were developed about this time (Chapter 9). 

Appropriate controls should always be run with any immuno-cytochemical procedure. Controls may include omitting the primary antibody, substituting preimmune serum, normal serum, or normal IgG for the primary antibody, adsorbing the primary antibody against the antigen, or immunostaining with an unrelated antibody. 

Appropriate controls should always be run with any immuno-cytochemical procedure. Controls may include omitting the... 

Rapkine s quantitative data are limited to the description of the fluctuation of soluble SH in the course of division. His proposal that the rise in soluble SH during the predivision stage is caused or at least accompanied by an increase in protein SH due to denaturation is not based on quantitative evidence and is not a logical consequence of the soluble SH picture. Nevertheless, it is the latter hypothesis that has attracted the most attention. The hypothesis led a number of workers to anticipate an increase, in connection vnth cell division, of total SH demonstrable by cytochemical procedures. Rapkine himself reported that he observed an increase in the intensity of the nitroprusside reaction during Phase II of the division of the sea-urchin egg and in certain Protozoa. 

Rocken C, Roessner A. An evaluation of antigen retrieval procedures for immu-noelectron microscopic classification of amyloid deposits. J. Histochem. Cytochem. 1999 47 1385-1394. 

Streefkerk JG, van der Ploeg M, van Duijn P. Agarose beads as matrices for proteins in cytophotometric investigations of immunohistoperoxidase procedures. /. Histochem. Cytochem. 1975 23 243-250. 

Martinez-Ramon, A., Knecht, E., Rubio, V., and Grisolia, S. (1990) Levels of carbamoyl phosphate synthetase I in livers of young and old rats assessed by activity and immunoassays and by electron microscopic immunogold procedures. J. Histochem. Cytochem. 38, 371-376. 

Finally, the localizations of low-molecular-weight compounds requires special specimen preparation techniques, as these compounds are often diffusible, water- or organic-solvent soluble, and solubilized by conventional fixation and dehydration procedures. The reader is referred to ref. (12) for the processing of cells and tissues for the cytochemical and histochemical localization of these compounds. 

Negata T. Electron microscope radioautography with cryo-fixation and dry mounting procedure. Acta Histochim Cytochem 1994 27 471 —489. 

Hsu SM, Raine L, Fanger H. 1981. Use of avidin-biotin-per-oxidase complex (ABC) in immunoperoxidase techniques a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 29 577-580. 

Hsu, S. M., Raine, L., and Fanger, H. (1981) The use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase technique a comparison between ABC and unlabeled antibody PAP procedures. J. Histochem. Cytochem. 29, 577-580. 

VanNoorden, S., Stuart, M. C., Cheung, A., Adams, E. F., andPolak, J. M. (1986) Localization of pituitary hormones by multiple immunoenzyme staining procedures using monoclonal and polyclonal antibodies. J. Histochem. Cytochem. 34, 287-292. 

Teclemariam-Mesbah R, Wortel J, Romijn HJ, Buijs RM. 1997. A simple silver-gold intensification procedure for double DAB labeling studies in electron microscopy. J Histochem Cytochem 45 619-621. 

Deitch, A D., Law, H., and DeVere White, R (1992) A stable propidium iodide staining procedure for flow cytometry J Histochem. Cytochem 30,967-972. 

The measurement of TSH was originally based on bioassays such as the stimulation of colloid droplet formation in the guinea pig thyroid gland and the release of labeled thyroidal iodide into mouse blood. These early in vivo bioassays, however, were of limited sensitivity and precision and were not applicable to the measurement of TSH in unfractionated serum. Most TSH bioassays have involved the in vitro stimulation of thyroid cyclic adenosine monophosphate (cAMP) or adenylate cyclase activity. The rat FRTL-5 thyroid cell line is an example of a particularly convenient and precise assay system. Unfortunately, such methods require purification and concentration of TSH from serum before assay. Sensitive detection of TSH in unfractionated serum is possible using a cytochemical bioassay, but this procedure is technically difficult and time-consuming. At present, immunoassay is the procedure of choice for the measurement of serum TSH in the clinical laboratory. 

Purification procedure via treatment with HCl Ultrastructural and cytochemical study Magnesium dependence and hair graying Study of dopachrome oxidoreductase and tyrosinase activity Correlation between melanin charge and intensity of ESR signal of irradiated hair... 

Straus W. Phenylhydrazine as inhibitor of horseradish peroxidase for use in immunoperoxidase procedures. J Histochem Cytochem. 1972 20 949. 

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