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Adipose Tissue Protocols by Gérard Ailhaud

By Gérard Ailhaud

Adipose tissue is now well-known as a widely-dispersed secretory organ that performs an important position in weight problems. Ailhaud (U. de Nice-Sophia Antipolis, great, France) and a group of ecu and US laboratory specialists and clinicians element the type, key thoughts hired, and their power difficulties, within the in vivo and in vitro research of brown and white fats tissues and cells from the dietary, physiological, and molecular degrees. significant remedy is accorded to the isolation, subcellular fractionation, and gene move into mature adipocytes, in addition to metabolic features. Illustrations contain microscopy learn effects, and a guy donning a conveyable detector for measuring adipose tissue blood circulate.

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1993) Application of the OsO4 maceration method to the study of human bioptic material. A procedure avoiding freeze-fracture. Microsc. Res. Tech. 26, 526–527. 37. Willison, J. H. M. and Rowe, A. T. (1980) Replica, shadowing and freeze-etching techniques, in Practical Methods in Electron Microscopy, vol. 8. (Glauert, A. ), North-Holland, Amsterdam, The Netherlands. 38. , and Björntorp P. (1985) Human brown adipose cells in culture. Exp. Cell Res. 159, 261–266. 39. , and Zancanaro, C. (1986) Ultrastructure of brown adipocytes mitochondria in cell culture from explants.

4, interscapular brown adipose tissue (IBAT) is carefully dissected under the microscope, weighed, and its volume measured by fluid displacement. 44 Cinti et al. 2. IBAT is cut into two parts along the middle plane of symmetry; each hemi-IBAT is paraffin-embedded and sectioned along parasagittal planes; 3-µm-thick serial sections are collected every millimeter and stained with H&E. 3. For each section, connective-vascular, unilocular, and multilocular areas are identified with a camera lucida, and measured with the Kontron image analyzer (45).

In our experience with AT, optimal sample size is about 2 cm3. Because of the texture of this tissue and to its high lipid content, all the parts exhibiting mechanical artifacts (compression and deformation caused by incorrect handling of forceps and knives) must be discarded. To perform this operation, we make a clean cut with a razorblade. 32 Cinti et al. 3. Dehydration and Embedding The final aim of these procedures is to obtain thin sections that can be stained and observed with a LM. To this end, after fixation, specimens must be embedded in a medium (paraffin) that will allow sectioning.

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