Prolong anti fade mounting media with dapi

Paraffin‐embedded MAT histopathological sections were de‐waxed and hydrated first, followed by the overnight hybridization step performed at 46 °C. EUB338 probe (5′‐GCTGCCTCCCGTAGGAGT‐3′, with Cy3 label) and specific probe designed for Klebsiella spp. (5′‐CCTACACACCAGCGTGCCA‐3′, with FITC label) were diluted to a final concentration of 5 ng µL⁻¹ in the hybridization buffer (20 mm Tris‐HCl, pH 7.4, 0.9 m NaCl, 0.1% SDS, and 35% formamide). After hybridization, the sections were rinsed with wash buffer (20 mm Tris‐HCl, pH 7.4, and 0.9 m NaCl) for three times (5 min/time), with Tris‐buffer for three times (3 min/time) at 48 °C and 1 min in ddH₂O. Dry the slides in an oven at 46 °C for 10 min, and then mounted the slides by Prolong anti‐fade mounting media with DAPI (Life Technologies). Microscopic observations were performed by Leica Laser Scanning confocal microscope (Leica TCS‐SP8, Leica Microsystems Inc, Buffalo Grove, IL, USA).