Next Article in Journal
Remifentanil Alleviates Propofol-Induced Burst Suppression without Affecting Bispectral Index in Female Patients: A Randomized Controlled Trial
Next Article in Special Issue
Mitochondrial DNA Variants in Patients with Liver Injury Due to Anti-Tuberculosis Drugs
Previous Article in Journal
“Third Window” and “Single Window” Effects Impede Surgical Success: Analysis of Retrofenestral Otosclerosis Involving the Internal Auditory Canal or Round Window
Previous Article in Special Issue
Clinical Characteristics and Treatment Outcomes of Definitive versus Standard Anti-Tuberculosis Therapy in Patients with Tuberculous Lymphadenitis
Open AccessArticle

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

1
Department of Pathology, Cell Biology and Biochemistry, Central Tuberculosis Research Institute, 2 Yauzskaya Alleya, 107564 Moscow, Russia
2
Faculty of Biology, Lomonosov Moscow State University, 1–12 Leninskie Gory, 119991 Moscow, Russia
3
Department of Comparative and Developmental Physiology, Koltsov Institute of Developmental Biology, Russian Academy of Sciences, 119334 Moscow, Russia
4
Department of Immunology, Central Tuberculosis Research Institute, 2 Yauzskaya Alleya, 107564 Moscow, Russia
5
Director of the Institute, Central Tuberculosis Research Institute, 2 Yauzskaya Alleya, 107564 Moscow, Russia
*
Author to whom correspondence should be addressed.
Authors contributed equally to this work.
J. Clin. Med. 2019, 8(8), 1185; https://doi.org/10.3390/jcm8081185
Received: 8 July 2019 / Revised: 30 July 2019 / Accepted: 2 August 2019 / Published: 7 August 2019
(This article belongs to the Special Issue Tuberculosis: Clinical Applications in the Diagnosis and Treatment)
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–Neelsen staining. Altogether, combining immunofluorescent labelling with the application of LSCM visualization significantly increases the effectiveness of M. tuberculosis detection. View Full-Text
Keywords: pulmonary tuberculosis; sensitivity of M. tuberculosis detection in tissue; Ziehl–Neelsen staining; confocal laser scanning microscopy; polyclonal antibody; immunohistochemistry pulmonary tuberculosis; sensitivity of M. tuberculosis detection in tissue; Ziehl–Neelsen staining; confocal laser scanning microscopy; polyclonal antibody; immunohistochemistry
Show Figures

Graphical abstract

MDPI and ACS Style

Erokhina, M.V.; Lepekha, L.N.; Voronezhskaya, E.E.; Nezlin, L.P.; Avdienko, V.G.; Ergeshov, A.E. Application of Laser Scanning Confocal Microscopy for the Visualization of M. tuberculosis in Lung Tissue Samples with Weak Ziehl–Neelsen Staining. J. Clin. Med. 2019, 8, 1185. https://doi.org/10.3390/jcm8081185

AMA Style

Erokhina MV, Lepekha LN, Voronezhskaya EE, Nezlin LP, Avdienko VG, Ergeshov AE. 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. 2019; 8(8):1185. https://doi.org/10.3390/jcm8081185

Chicago/Turabian Style

Erokhina, Maria V.; Lepekha, Larisa N.; Voronezhskaya, Elena E.; Nezlin, Leonid P.; Avdienko, Vadim G.; Ergeshov, Atadzhan E. 2019. "Application of Laser Scanning Confocal Microscopy for the Visualization of M. tuberculosis in Lung Tissue Samples with Weak Ziehl–Neelsen Staining" J. Clin. Med. 8, no. 8: 1185. https://doi.org/10.3390/jcm8081185

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Search more from Scilit
 
Search
Back to TopTop