Figure 6: Biochemical characterization of the composition of the ECM of Mtb biofilms.
From: Thiol reductive stress induces cellulose-anchored biofilm formation in Mycobacterium tuberculosis
Mtb biofilms overexpressing GFP were developed in standing cultures (without Tween-80) by exposure to TRS and then stained with specific fluorophores: SYPRO Ruby for staining the proteins (a), Nile red (1 mg ml−1) for staining the lipids (b), Texas red (0.5 mg ml−1) for staining polysaccharides (c), lectin ConA (200 μm) for staining the α-mannopyranosyl- and α-glucopyranosyl-rich polysaccharides (d), and PI (15 μM) for staining the extracellular DNA (e). Cultures were then analysed using CLSM. (i) The overlay image of ii (GFP bacteria) and iii (showing the specific staining pattern). (vi) The overlay image in three dimensions for iv (three-dimensional image of the biofilm as traced with GFP-overexpressing bacteria) and v (specific staining of the biofilm in three dimensions).
