Application of Laser Scanning Confocal Microscopy for the Visualization of M. tuberculosis in Lung Tissue Samples with Weak Ziehl-Neelsen Staining

Journal of Clinical Medicine
Maria V ErokhinaAtadzhan E Ergeshov

Abstract

One of the key requirements for the diagnosis of pulmonary tuberculosis is the identification of M. tuberculosis in tissue. In this paper, we present the advantages of specific fluorescent antibody labelling, combined with laser scanning confocal microscopy (LSCM), for the detection of M. tuberculosis in histological specimens of lung tissues. We demonstrate that the application of LSCM allows: (i) The automatic acquisition of images of the whole slice and, hence, the determination of regions for subsequent analysis; (ii) the acquisition of images of thick (20-40 μm) slices at high resolution; (iii) single bacteria identification; and (iv) 3D reconstruction, in order to obtain additional information about the distribution, size, and morphology of solitary M. tuberculosis; as well as their aggregates and colonies, in various regions of tuberculosis inflammation. LSCM allows for the discrimination of the non-specific fluorescence of bacteria-like particles and their aggregates presented in histological lung samples, from the specific fluorescence of labelled M. tuberculosis, using spectrum emission analysis. The applied method was effective in the identification of M. tuberculosis in lung histological samples with weak Ziehl-Neel...Continue Reading

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Citations

May 4, 2021·Frontiers in Microbiology·Julio FloresLeslie Chavez-Galan

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Methods Mentioned

BETA
PCR
confocal microscopy
light microscopy
Fluorescence
biopsy
light
fluorescence microscopy
fluorescencemicroscopy

Software Mentioned

STATISTICA
ImageJ
Excel
PhotoShop CC

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