Tuberculosis remains one of the major infectious diseases in the world. Current vaccine testing relies on in vivo models including the mouse aerosol challenge model. To determine structural changes in the 3D of granulomas caused by Mycobacterium tuberculosis previous observations have relied on serial sectioning with histopathology techniques. However, this has several disadvantages including the possibility of missing or distorting information in the 3D.

Optical clearing techniques may offer a way around this by making the whole organs transparent using chemical procedures, and then imaging by confocal laser scanning microscopy to produce optical sections. Therefore this study aimed to see whether optical clearing techniques can be used to study granulomas in 3D and the structural changes to the surrounding tissue. Results to date have shown that it is possible to optically clear lobes of the mouse lung and to resolve individual nuclei. We now hope to use the same techniques to image M. tuberculosis infected lung and to compare with other organs.

Figures:

Tile scan of a section of the mid-lobe of the mouse lung. The 3D image has been cropped to show a cross section through the alveoli. Acridine orange staining showing the nuclei of the lung. Images taken on a Leica SP8 X CLSM using a multiphoton (900nm) and reflective non-descanned detectors. Pseudo-colouring based on depth through the tissue.

Zoomed in image of figure 1. Individual nuclei (green) can be seen in the image. Scale bar = 100µm

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EMC Abstracts - https://emc-proceedings.com/abstract/3-dimensional-measurements-of-tuberculosis-granulomas-using-optical-clearing-techniques/