Slicing overcomes the bacterial cell wall barrier to fluorescence <i>in situ</i> hybridization.

ABSTRACT: Simultaneous visualization of all bacterial species in a polymicrobial biofilm by fluorescence in situ hybridization (FISH) remains a challenge because bacterial taxa respond differently to the hybridization procedure. This heterogeneity is due, in part, to the cell wall barrier of Gram-positive taxa. Enzymatic procedures required to permeabilize the cell walls of Gram-positive microbes can result in the disruption or loss of Gram-negative bacteria. Here, we demonstrate a procedure that enables the hybridization of difficult-to-hybridize bacteria while preserving the microarchitecture of all bacteria within a biofilm. The key feature of the procedure is to physically cut through the cell wall, thus allowing probe entry. We first embed the sample in a covalently crosslinked, glycol m

SUBMITTER: Gundrum J 

PROVIDER: S-EPMC12772237 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature