Single-cell microfluidics is a powerful method to study bacteria and determine their susceptibility to antibiotic treatment. Glass treatment by adhesive molecules is a potential solution to immobilize bacterial cells and perform microscopy, but traditional cationic polymers such as polylysine deeply affect bacterial physiology. In this work, we chemically characterized a class of chitosan polymers for their biocompatibility when adsorbed to glass. Chitosan chains of known length and composition allowed growth of Escherichia coli cells without any deleterious effects on cell physiology. Combined with a machine learning approach, this method could measure the antibiotic susceptibility of a diversity of clinical strains in less than 1 h and with higher accuracy than current methods. Finally, chitosan polymers also supported growth of Klebsiella pneumoniae, another bacterial pathogen of clinical significance. IMPORTANCE Current microfluidic techniques are powerful to study bacteria and determine their response to antibiotic treatment, but they are currently limited by their complex manipulation. Chitosan films are fully biocompatible and could thus be a viable replacement for existing commercial devices that currently use polylysine. Thus, the low cost of chitosan slides and their simple implementation make them highly versatile for research as well as clinical use.
Reaction in gel between the sodium salt of 1,4-phenylenediacetic acid (Na2C10O4H8–Na2 p-pda) and lanthanum chloride yields single crystals of the three-dimensional coordination polymer poly[[tetraaquatris(μ-1,4-phenylenediacetato)dilanthanum(III)] octahydrate], {[La2(C10H8O4)3(H2O)4]·8H2O}∞. The LaIII coordination polyhedron can be described as a slightly distorted monocapped square antiprism. One of the two p-pda2− ligands is bound to four LaIII ions and the other to two LaIII ions. Each LaIII atom is coordinated by five ligands, thereby generating a metal–organic framework with potential porosity properties.
31Single cell microfluidics is powerful to study bacteria and determine their susceptibility to 32 antibiotics treatment. Glass treatment by adhesive molecules is a potential solution to 33 immobilize bacterial cells and perform microscopy but traditional cationic polymers such as 34 poly-lysine deeply affect bacterial physiology. In this work, we chemically characterized a class 35 of chitosan polymers for their biocompatibility when adsorbed to glass. Chitosan chains of 36 known length and composition allowed growth of Escherichia coli cells without any deleterious 37 effects on cell physiology. Combined with a machine-learning approach, this method could 38 measure the antibiotics susceptibility of a diversity of clinical strains in less than 1 hour and 39with higher accuracy than current methods. Last, chitosan polymers also supported growth of 40Klebsiella pneumoniae, another bacterial pathogen of clinical significance. The low cost of 41 chitosan slides and their simple implementation makes them highly versatile for research as 42well as clinical use. 43 44 65 this approach does not allow rapid changes of the medium or injection of 66 chemicals and thus the kinetics and precise dose-dependent effects are poorly 67controlled ( 3, 4). 68Alternative methods have remedied these issues by growing bacteria immediately in contact 69with a glass surface. Because most bacteria do not directly adhere to glass, immobilization 70procedures are required, which include direct physical immobilization of the bacteria in micro 71 channels or glass functionalization by adhesive polymers. The use of micro-channels is 72certainly compatible with HEM and it allows fast AST with high accuracy (2, 5). However, this 73 method requires expert handling, complex nanolithography to produce the channels and 74 extensive development to be used for the study of a given bacterial species. Alternatively, 75bacterial adhesion on glass can be obtained by functionalizing a glass slide with adhesive 76 polymers/molecules. This approach can also be difficult because the polymer must be fully 77 biocompatible and the functionalization procedure and surface chemistry can be complex. 78Indeed, although this approach has been widely used for eukaryotic cells, the choice for 79polymers biocompatible with bacteria is limited. Cationic polymers such as poly-lysine bind 80 glass surfaces effectively and promote adhesion of a wide range of bacterial species. However, 81poly-lysine also generates cell envelope stress and has been shown to dissipate/diminish the 82 membrane potential in several Gram negative of Gram positive species (e.g. Escherichia coli 83or Bacillus subtilis (6)(7)(8)). For clinical microbiology applications, this issue is particularly 84 sensitive because changes in the membrane potential can directly affect antimicrobial 85 susceptibility (9) and thus produce false negative or even worst, false positive results in AST. 86Thus, there is a need in developing new functional polymers with neutral effects on bacterial 87 physiology for si...
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