Preparation of Tissues

A chloric acid digestion was used by Backer 2 391 for the preparation of tissue samples. The digest is simply diluted to determine iron, zinc, and copper. The tantalum sampling boat technique was used by Emmermann and Luecke 2531 to measure lead, zinc, and silver in prepared soil solutions. White 1S81 treated ashed plants with hydroxylamine in IN hydrochloric acid to reduce and dissolve oxides of manganese, prior to its determination by atomic absorption spectroscopy. 

Preparation of data forms and books. Preparation of tissue slides and microscopic evaluation of these slides ... 

Stadie, W.C. and Riggs, B.C. (1944). Microtome for the preparation of tissue slices for metabolic studies of tissues in vitro. J. Biol. Chem. 154 687-690. 

Transfer the deparaffmized, rehydrated, and methanolic-peroxide blocked (optional) slides (see Notes 2-6) into plastic Coplin jars or containers filled with HIER buffer (see Note 7). (See Chapter 12 for preparation of tissue sections.)... 

To prepare lysates from (nonstimulated) fibroblasts, cells from one confluent 78-cm2 plate are suspended in 0.15 ml lysis buffer (see below) and lysed by freezing and thawing six times and subsequent centrifugation at 13,000 x for 5 min. An aliquot of 0.05 ml of the supernatant is directly used for the enzyme assay. The preparation of tissue homogenate is described in section 6.1.4.1. GTP cyclohydrolase I, subheading Specimen . 

A different procedure for the SCM confirmation was also published based on atmospheric pressure ionization with a collision-induced dissociation. The triple-quadrupole mass spectrometer operated in the Q1-Q3 product-ion mode. The preparation of tissue samples is described in the next section (111). 

Many of the contradictory results are possibly occasioned by use of different methods of assay and of different preparations of tissue. The presence of endogenous co-factors and inhibitors has only recently been considered. 

Preparation of Tissues Tissues are quick-frozen in OCT embedding media, and 4 to 6 pm cryosections are placed on slides and allowed to dry, or stored at 4 °C for a few days or at -80 °C for longer terms prior to fixation. Prolonged retention of tissue sections is usually to be avoided even at -80 °C as antigen deterioration can occur even under these conditions. Blood smears are prepared fresh and allowed to dry prior to fixation.Tissue preparation (sectioning, drying, fixation) is governed by the requirements needed to preserve the... 

Krumdieck CL, Dos Santos JE, Ho K-J (1980) A new instrument for the rapid preparation of tissue slices. Anal Biochem 104 118-123... 

Any proteomic study starts with the collection of proteins from biological samples such as cell culture media, cultured cells, serum, or any biological fluid, and a variety of animal tissues. The first step is to obtain a protein sample under conditions of least protein degradation. This involves use of various protease inhibitors that stop the protein degradation. The use of protease inhibitors depends on the type of sample and the analytical technique used in the subsequent analysis. The selection of protease inhibitors used is critical since many protease inhibitors and detergents used in the preparation of tissue homogenates can interfere with mass spectrometry (MS)... 

Although a considerable literature records numerous studies in this field (A19, B17, C12, J3, L9, P16, S4, S5), many have been performed on preparations of tissues from other than human sources. In conformity with the subject of this chapter and to avoid species differences, most attention will be directed to human tissue alkaline phosphatases and in particular their variants. The stereospecific L-phenylalanine inhibition has provided the impetus to study its molecular mechanism, which necessarily requires an understanding of the mechanism of catalysis. It is expected that discovery of other stereospecific inhibitors will follow and that they may have even greater utility than L-phenylalanine. However, since it is the first such unique inhibitor, this section of the chapter will receive extensive treatment after a consideration of some basic kinetic information. 

This step includes many stages to prepare tissue sections for the hybridization. Initial preparing of tissue sections is the same as described for immunohistochemistry. 

An essential requirement is the refrigerated centrifuge capable of generating forces from approximately 1000-50,000. Appropriate and dependable instruments for this purpose include the DuPont Sorvall RC-5 and Beckman J2-21 series of instruments Because the centrifuge is used both in the preparation of tissues and, not infrequently, to terminate the binding reaction, it should be equipped with rotors capable of handling a small number of large samples ( 50 mL) and multiple samples of smaller volumes. 

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