Dataset Information

From macro to micro: a combined bioluminescence-fluorescence approach to monitor bacterial localization.

ABSTRACT: Bacterial bioluminescence is widely used to study the spatiotemporal dynamics of bacterial populations and gene expression in vivo at a population level but cannot easily be used to study bacterial activity at the level of individual cells. In this study, we describe the development of a new library of mini-Tn7-lux and lux::eyfp reporter constructs that provide a wide range of lux expression levels, and which combine the advantages of both bacterial bioluminescence and fluorescent proteins to bridge the gap between macro- and micro-scale imaging techniques. We demonstrate that a dual bioluminescence-fluorescence approach using the lux operon and eYFP can be used to monitor bacterial movement in plants both macro- and microscopically and demonstrate that Pseudomonas syringae pv phaseolicola can colonize the leaf vascular system and systemically infect leaves of common bean (Phaseolus vulgaris). We also show that bacterial bioluminescence can be used to study the impact of plant immune responses on bacterial multiplication, viability and spread within plant tissues. The constructs and approach described in this study can be used to study the spatiotemporal dynamics of bacterial colonization and to link population dynamics and cellular interactions in a wide range of biological contexts.

SUBMITTER: Soldan R

PROVIDER: S-EPMC8614114 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature

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From macro to micro: a combined bioluminescence-fluorescence approach to monitor bacterial localization.

Soldan Riccardo R Sanguankiattichai Nattapong N Bach-Pages Marcel M Bervoets Indra I Huang Wei E WE Preston Gail M GM

Environmental microbiology 20210122 4

Bacterial bioluminescence is widely used to study the spatiotemporal dynamics of bacterial populations and gene expression in vivo at a population level but cannot easily be used to study bacterial activity at the level of individual cells. In this study, we describe the development of a new library of mini-Tn7-lux and lux::eyfp reporter constructs that provide a wide range of lux expression levels, and which combine the advantages of both bacterial bioluminescence and fluorescent proteins to br ...[more]

PMID: 33103833

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Project description:Inertial measurement units (IMU) are proven as efficient tools for swimming analysis by overcoming the limits of video-based systems application in aquatic environments. However, coaches still believe in the lack of a reliable and easy-to-use analysis system for swimming. To provide a broad view of swimmers' performance, this paper describes a new macro-micro analysis approach, comprehensive enough to cover a full training session, regardless of the swimming technique. Seventeen national level swimmers (5 females, 12 males, 19.6 ± 2.1 yrs) were equipped with six IMUs and asked to swim 4 × 50 m trials in each swimming technique (i.e., frontcrawl, breaststroke, butterfly, and backstroke) in a 25 m pool, in front of five 2-D cameras (four under water and one over water) for validation. The proposed approach detects swimming bouts, laps, and swimming technique in macro level and swimming phases in micro level on all sensor locations for comparison. Swimming phases are the phases swimmers pass from wall to wall (wall push-off, glide, strokes preparation, swimming, and turn) and micro analysis detects the beginning of each phase. For macro analysis, an overall accuracy range of 0.83-0.98, 0.80-1.00, and 0.83-0.99 were achieved, respectively, for swimming bouts detection, laps detection and swimming technique identification on selected sensor locations, the highest being achieved with sacrum. For micro analysis, we obtained the lowest error mean and standard deviation on sacrum for the beginning of wall-push off, glide and turn (-20 ± 89 ms, 4 ± 100 ms, 23 ± 97 ms, respectively), on shank for the beginning of strokes preparation (0 ± 88 ms) and on wrist for the beginning of swimming (-42 ± 72 ms). Comparing the swimming techniques, sacrum sensor achieves the smallest range of error mean and standard deviation during micro analysis. By using the same macro-micro approach across different swimming techniques, this study shows its efficiency to detect the main events and phases of a training session. Moreover, comparing the results of both macro and micro analyses, sacrum has achieved relatively higher amounts of accuracy and lower mean and standard deviation of error in all swimming techniques.

Dataset Information

From macro to micro: a combined bioluminescence-fluorescence approach to monitor bacterial localization.

Publications

From macro to micro: a combined bioluminescence-fluorescence approach to monitor bacterial localization.

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