Flow cytometry(FCM)is a powerful technique for single-bacteria analysis via simultaneous light-scattering and fluorescence measurements.By offering high-throughput,quantitative,and multiparameter analysis at the singl...Flow cytometry(FCM)is a powerful technique for single-bacteria analysis via simultaneous light-scattering and fluorescence measurements.By offering high-throughput,quantitative,and multiparameter analysis at the single-cell level,FCM has gained an increased popularity in microbiological research,food safety monitoring,water quality control,and clinical diagnosis.Here we will review the recent applications of flow cytometry in areas such as(1)total bacterial cell count,(2)bacterial viability analysis,(3)specific bacterial detection and identification,(4)characterization of physiological changes under environmental perturbations,and(5)biological function studies.Nevertheless,despite these widespread applications,challenges still remain for the detection of small sizes of bacteria and biochemical features that cannot be brightly stained via fluorescence.Recent improvement in FCM instrumentation will be discussed,and particularly the development of high sensitivity flow cytometry for advanced analysis of single bacterial cells will be highlighted.展开更多
Hydrophilic polymers are very useful in biomedical applications.In this study,biocom-patible polyethylene glycol(PEG)and polyvinylpyrrolidone(PVP)polymers end-capped with succinimidyl groups were either modified or sy...Hydrophilic polymers are very useful in biomedical applications.In this study,biocom-patible polyethylene glycol(PEG)and polyvinylpyrrolidone(PVP)polymers end-capped with succinimidyl groups were either modified or synthesised and attached to poly-vinylchloride surfaces.The modified surfaces were evaluated with cell adhesion and bacterial adhesion.3T3 mouse fibroblast cells and three bacteria species were used to evaluate surface adhesion activity.Results showed that the modified surface exhibited significantly reduced 3T3 cell adhesion with a 50%-69%decrease for PEG and a 64%-81%for PVP,as compared to unmodified polyvinylchloride.The modified surface also showed significantly reduced bacterial attachment with 22%-78%,18%-76%and 20%-75%decrease for PEG and 22%-76%,18%-76%and 20%-73%for PVP to Staphy-lococcus aureus,Escherichia coli and Pseudomonas aeruginosa,respectively,as compared to unmodified polyvinylchloride.It seems that an appropriate chain length or molecular weight(neither the longest nor the shortest chain length)determines the lowest cell and bacterial adhesion in terms of PEG.On the other hand,a mixture of polymers with different chain lengths exhibited the lowest cell and bacterial adhesion in terms of PVP.展开更多
Rainwater contains substantial bacteria and rain is an efficient pathway for the dissemination of bacteria from the atmosphere to land and water surfaces.However,quantitative information on rainwater bacteria is very ...Rainwater contains substantial bacteria and rain is an efficient pathway for the dissemination of bacteria from the atmosphere to land and water surfaces.However,quantitative information on rainwater bacteria is very limited due to the lack of a reliable method.In this study,the epifluorescence microscopy enumeration with the LIVE/DEAD BacLight Bacterial Viability Kit stain was verified to quantify the abundance of viable and non-viable bacterial cells in rainwater,with the 4＇,6-diamidino-2-phenylindole（DAPI） stain for the reference of total cell counts.Results showed that the total counts of bacterial cells by LIVE/DEAD BacLight staining were consistent with those by DAPI staining,and the average detection efficiency was（109 ± 29）%.The ratio of cell count with glutaraldehyde fixation to that without fixation was（106 ± 5）%on average.The bacterial concentration in negative control was usually an order of magnitude lower than that in rainwater samples.However,in case of small precipitation,the abundance in negative control could be more than that in rainwater samples.These results indicate that the enumeration with LIVE/DEAD BacLight bacterial viability assay coupled with glutaraldehyde fixation and careful negative control investigation is an approach applicable to the measurement of the concentration and viability of bacterial cells in rainwater.展开更多
The study aims to identify the potential acute effects of suspended aluminum nitride（Al N）nanoparticles（NPs） on soluble microbial products（SMP） of activated sludge.Cultured activated sludge loaded with 1,10,50,1...The study aims to identify the potential acute effects of suspended aluminum nitride（Al N）nanoparticles（NPs） on soluble microbial products（SMP） of activated sludge.Cultured activated sludge loaded with 1,10,50,100,150 and 200 mg/L of Al N NPs were carried out in this study.As results showed,Al N NPs had a highly inverse proportionality to bacterial dehydrogenase and OUR,indicating its direct toxicity to the activated sludge viability.The toxicity of Al N NPs was mainly due to the nano-scale of Al N NPs.In SMP,Al N NPs led to the decrease of polysaccharide and humic compounds,but had slight effects on protein.The decrease of tryptophan-like substances in SMP indicated the inhibition of Al N NPs on the bacterial metabolism.Additionally,Al N NPs reduced obviously the molecular weight of SMP,which might be due to the nano-scale of Al N.展开更多
基金the National Key Basic Research Program of China(2013CB933703)the National Natural Science Foundation of China(91313302,21105082,21225523,21472158,21027010,21521004)the Program for Changjiang Scholars and Innovative Research Team in University(IRT13036)
文摘Flow cytometry(FCM)is a powerful technique for single-bacteria analysis via simultaneous light-scattering and fluorescence measurements.By offering high-throughput,quantitative,and multiparameter analysis at the single-cell level,FCM has gained an increased popularity in microbiological research,food safety monitoring,water quality control,and clinical diagnosis.Here we will review the recent applications of flow cytometry in areas such as(1)total bacterial cell count,(2)bacterial viability analysis,(3)specific bacterial detection and identification,(4)characterization of physiological changes under environmental perturbations,and(5)biological function studies.Nevertheless,despite these widespread applications,challenges still remain for the detection of small sizes of bacteria and biochemical features that cannot be brightly stained via fluorescence.Recent improvement in FCM instrumentation will be discussed,and particularly the development of high sensitivity flow cytometry for advanced analysis of single bacterial cells will be highlighted.
文摘Hydrophilic polymers are very useful in biomedical applications.In this study,biocom-patible polyethylene glycol(PEG)and polyvinylpyrrolidone(PVP)polymers end-capped with succinimidyl groups were either modified or synthesised and attached to poly-vinylchloride surfaces.The modified surfaces were evaluated with cell adhesion and bacterial adhesion.3T3 mouse fibroblast cells and three bacteria species were used to evaluate surface adhesion activity.Results showed that the modified surface exhibited significantly reduced 3T3 cell adhesion with a 50%-69%decrease for PEG and a 64%-81%for PVP,as compared to unmodified polyvinylchloride.The modified surface also showed significantly reduced bacterial attachment with 22%-78%,18%-76%and 20%-75%decrease for PEG and 22%-76%,18%-76%and 20%-73%for PVP to Staphy-lococcus aureus,Escherichia coli and Pseudomonas aeruginosa,respectively,as compared to unmodified polyvinylchloride.It seems that an appropriate chain length or molecular weight(neither the longest nor the shortest chain length)determines the lowest cell and bacterial adhesion in terms of PEG.On the other hand,a mixture of polymers with different chain lengths exhibited the lowest cell and bacterial adhesion in terms of PVP.
基金supported by a Grant-in-Aid for Challenging Exploratory Research(No.15K12192)from the Japan Society for the Promotion of Science(JSPS)The State Scholarship Fund of Chinese Scholarship Council(CSC No.201406010350)provides Mr.Wei Hu the scholarship for his study and stay at the Prefectural University of Kumamoto
文摘Rainwater contains substantial bacteria and rain is an efficient pathway for the dissemination of bacteria from the atmosphere to land and water surfaces.However,quantitative information on rainwater bacteria is very limited due to the lack of a reliable method.In this study,the epifluorescence microscopy enumeration with the LIVE/DEAD BacLight Bacterial Viability Kit stain was verified to quantify the abundance of viable and non-viable bacterial cells in rainwater,with the 4＇,6-diamidino-2-phenylindole（DAPI） stain for the reference of total cell counts.Results showed that the total counts of bacterial cells by LIVE/DEAD BacLight staining were consistent with those by DAPI staining,and the average detection efficiency was（109 ± 29）%.The ratio of cell count with glutaraldehyde fixation to that without fixation was（106 ± 5）%on average.The bacterial concentration in negative control was usually an order of magnitude lower than that in rainwater samples.However,in case of small precipitation,the abundance in negative control could be more than that in rainwater samples.These results indicate that the enumeration with LIVE/DEAD BacLight bacterial viability assay coupled with glutaraldehyde fixation and careful negative control investigation is an approach applicable to the measurement of the concentration and viability of bacterial cells in rainwater.
基金supported by the National Natural Science Foundation of China(No.51378368)the Foundation of State Key Laboratory of Pollution Control and Resource Reuse(Tongji University),China(No.PCRRT16003)Shenzhen Science and Technology Research Fund(No.CXZZ20150330151321966)
文摘The study aims to identify the potential acute effects of suspended aluminum nitride（Al N）nanoparticles（NPs） on soluble microbial products（SMP） of activated sludge.Cultured activated sludge loaded with 1,10,50,100,150 and 200 mg/L of Al N NPs were carried out in this study.As results showed,Al N NPs had a highly inverse proportionality to bacterial dehydrogenase and OUR,indicating its direct toxicity to the activated sludge viability.The toxicity of Al N NPs was mainly due to the nano-scale of Al N NPs.In SMP,Al N NPs led to the decrease of polysaccharide and humic compounds,but had slight effects on protein.The decrease of tryptophan-like substances in SMP indicated the inhibition of Al N NPs on the bacterial metabolism.Additionally,Al N NPs reduced obviously the molecular weight of SMP,which might be due to the nano-scale of Al N.
