Immunohistochemical detection of Mycobacterium tuberculosis in tissues of consumptives using laser scanning microscopy

Resumo

A simple and reliable method for detection of Mycobacterium tuberculosis (MBT) in histological specimens of lung tissue of consumptives using obtained anti-MBT polyclonal antibodies and laser scanning confocal microscopy was proposed. The method detected both colonies and single MBT and localized intraand extracellularly in caseous necrosis areas and perifocal regions. It was noted that automatic image capture of a large amount of tissue (up to 2 × 2 × 0.1 mm) and subsequent combination of optical sections in a threedimensional image provided the opportunity to carry out automated search and counting of MBT in tissues. Registration of the emission spectrum was allowed separating specific signals from nonspecific fluorescence in specimens.