DUAL-CHANNEL LASER-SCANNING MICROSCOPY FOR THE IDENTIFICATION AND QUANTIFICATION OF PROLIFERATING SKELETAL-MUSCLE SATELLITE CELLS FOLLOWINGSYNERGIST ABLATION

Proliferating skeletal muscle satellite cells are the source of additi onal myonuclei which allow skeletal muscle to grow and regenerate. Pre viously, proliferating satellite cells were identified in situ by tech niques which were limited either by tissue processing time or inabilit y to observe complete muscle sections, or by errors made in separating these cells from proliferating nonmyogenic cells. To overcome these p roblems a new method has been devised for the identification and quant ification of proliferating satellite cells in situ by light microscopy . The technique involves dual-channel laser scanning imaging of whole muscle sections for the localisation of both the muscle fibre basal la mina and the cell division marker bromodeoxyuridine. Using this techni que satellite cell proliferation was quantified in mouse limb muscle f ollowing synergist ablation. Dual-channel laser scanning microscopy al lowed precise localisation of proliferating satellite cells in the exp erimental model and, after 4 d, synergist ablation was shown to have p roduced significant satellite cell proliferation when compared with co ntralateral and sham-operated controls.