Histochemical study of lipids in the tissues

When processing the data take into account the high lipid solubility in organic solvents. So use slices on a freezing microtome. Fill in paraffin and celloidin is made only when using those methods of detection of L. in which their preliminary translate into insoluble condition (for example, the influence of potassium bichromate). As preservative used formalin. To save phosphatides in the formalin solution it is recommended to add 1 % of calcium chloride. If necessary, the material can be filled in gelatine. Stained sections conclude in glycerin, glycerine-gelatine and fructose syrup.
Most often use the following methods of detection HP Color stanowimy dyes (Sudan III, sarlah red, Sudan black) is made for the total of detection of lipids; the specificity of the method generally accepted. For determination of free fatty acids applies color Neel-blousonform in which fatty acids are painted in blue and the neutral fat - in pink, and the reaction of filler, allowing to receive blue colouring fatty acids (neutral fat is not painted). Unsaturated fatty acids, along with other unsaturated compounds can be identified by the method Smith - Dietrich. Less tested to determine the fatty acids method Gomory with the use of lead salts and ammonium sulfide and recommendation Okamoto (K. Okamoto) to use in the reaction of fishler not hematoxylin and para-dimethyl-amino-benzel-roganin. Reactions Schiff and Govinda can detect the products of oxidative transformation of neutral fat - aldehydes and peroxide. Method Baker - a reliable way of identifying phosphatides. You can also use the methods of Cacho and Minibike, however, they, apparently, are less reliable. Pearce (A. Pearse ) proposes to use to identify phosphatides copper phthalocyanines. Reaction Shultze to identify cholesterol and its esters most common. Free cholesterol is well detected by reaction with digitonin.
To determine the nature detected lipid great importance selective extraction of individual groups of lipids those or other solvents. The scheme sequential extraction small samples with okraski stanowimy dyes [Calig (J. Keilig)]. Okamoto etc. are offered method differentiated definition of phosphatides, cerebrosides, sphingomyelin and lecithin and separate sphingomyelin using nitrate mercury and processing diphenylcarbazone. In all cases, histochemical detection of L. it is appropriate to control lipid nature of detected substances. As a solvent, which removes all HP, recommended heated mixture (1:1) methyl alcohol And chloroform.