Bacterial resistance,primarily stemming from misdiagnosis,misuse,and overuse of antibacterial medications in humans and animals,is a pressing issue.To address this,we focused on developing a fluorescent probe for the ...Bacterial resistance,primarily stemming from misdiagnosis,misuse,and overuse of antibacterial medications in humans and animals,is a pressing issue.To address this,we focused on developing a fluorescent probe for the detection of bacteria,with a unique feature--an exceptionally long fluorescence lifetime,to overcome autofluorescence limitations in biological samples.The polymyxin-based probe(ADOTA-PMX)selectively targets Gram-negative bacteria and used the red-emitting fluorophore azadioxatriangulenium(with a reported fluorescence lifetime of 19.5 ns).Evaluation of ADOTA-PMX’s bacterial labeling efficacy revealed strong specificity for Gram-negative bacteria,and full spectral fluorescence lifetime imaging microscopy demonstrated the potential of ADOTA-PMX for bacterial imaging applications.The probe exhibited a lifetime of 4.5 ns when bound to bacteria,possibly indicating interactions with the bacterial outer membrane.Furthermore,the fluorescence lifetime measurements of ADOTA-PMX labeled bacteria could be performed using a benchtop fluorimeter without the need of sophisticated microscopes.This study represents the first targeted probe for fluorescence lifetime imaging,offering sensitivity for detecting Gram-negative bacteria and enabling multiplexing via fluorescence lifetime imaging.展开更多
基金funded by Engineering and Physical Sciences Research Council(EPSRC,United Kingdom)Interdisciplinary Research Collaboration,Grant Nos.EP/K03197X/1 and EP/R005257/1funded by a Royal Society,Grant No.RGS\R1\201163+1 种基金funded via the BBSRC EastBIO doctoral training program,grant BB/M010996/1funded by a Newton-Bhabha International Fellowship from The Royal Society,UK and The Science and Engineering Research Board,India,Grant No.NIF/R1/192688.
文摘Bacterial resistance,primarily stemming from misdiagnosis,misuse,and overuse of antibacterial medications in humans and animals,is a pressing issue.To address this,we focused on developing a fluorescent probe for the detection of bacteria,with a unique feature--an exceptionally long fluorescence lifetime,to overcome autofluorescence limitations in biological samples.The polymyxin-based probe(ADOTA-PMX)selectively targets Gram-negative bacteria and used the red-emitting fluorophore azadioxatriangulenium(with a reported fluorescence lifetime of 19.5 ns).Evaluation of ADOTA-PMX’s bacterial labeling efficacy revealed strong specificity for Gram-negative bacteria,and full spectral fluorescence lifetime imaging microscopy demonstrated the potential of ADOTA-PMX for bacterial imaging applications.The probe exhibited a lifetime of 4.5 ns when bound to bacteria,possibly indicating interactions with the bacterial outer membrane.Furthermore,the fluorescence lifetime measurements of ADOTA-PMX labeled bacteria could be performed using a benchtop fluorimeter without the need of sophisticated microscopes.This study represents the first targeted probe for fluorescence lifetime imaging,offering sensitivity for detecting Gram-negative bacteria and enabling multiplexing via fluorescence lifetime imaging.
