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Enhancing fluorescent protein photostability through robot-assisted photobleaching.

ABSTRACT: Improving fluorescent proteins through the use of directed evolution requires robust techniques for screening large libraries of genetic variants. Here we describe an effective and relatively low-cost system for screening libraries of fluorescent protein variants for improved photostability in the context of colonies on a Petri dish. Application of this system to the yellow fluorescent protein mCitrine, led to the development of Citrine2 with improved photostability and similar high fluorescent brightness. The photobleaching robot was constructed using a Lego Mindstorms Ev3 set and a xenon arc lamp, which together create even and high irradiance over an entire Petri dish through patterned illumination.

SUBMITTER: Wiens MD

PROVIDER: S-EPMC6047911 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

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Enhancing fluorescent protein photostability through robot-assisted photobleaching.

Wiens M D MD Hoffmann F F Chen Y Y Campbell R E RE

Integrative biology : quantitative biosciences from nano to macro 20180701 7

Improving fluorescent proteins through the use of directed evolution requires robust techniques for screening large libraries of genetic variants. Here we describe an effective and relatively low-cost system for screening libraries of fluorescent protein variants for improved photostability in the context of colonies on a Petri dish. Application of this system to the yellow fluorescent protein mCitrine, led to the development of Citrine2 with improved photostability and similar high fluorescent ...[more]

PMID: 29897363

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Project description:Proprioception has a crucial role in promoting or hindering motor learning. In particular, an intact position sense strongly correlates with the chances of recovery after stroke. A great majority of neurological patients present both motor dysfunctions and impairments in kinesthesia, but traditional robot and virtual reality training techniques focus either in recovering motor functions or in assessing proprioceptive deficits. An open challenge is to implement effective and reliable tests and training protocols for proprioception that go beyond the mere position sense evaluation and exploit the intrinsic bidirectionality of the kinesthetic sense, which refers to both sense of position and sense of movement. Modulated haptic interaction has a leading role in promoting sensorimotor integration, and it is a natural way to enhance volitional effort. Therefore, we designed a preliminary clinical study to test a new proprioception-based motor training technique for augmenting kinesthetic awareness via haptic feedback. The feedback was provided by a robotic manipulandum and the test involved seven chronic hemiparetic subjects over 3?weeks. The protocol included evaluation sessions that consisted of a psychometric estimate of the subject's kinesthetic sensation, and training sessions, in which the subject executed planar reaching movements in the absence of vision and under a minimally assistive haptic guidance made by sequences of graded force pulses. The bidirectional haptic interaction between the subject and the robot was optimally adapted to each participant in order to achieve a uniform task difficulty over the workspace. All the subjects consistently improved in the perceptual scores as a consequence of training. Moreover, they could minimize the level of haptic guidance in time. Results suggest that the proposed method is effective in enhancing kinesthetic acuity, but the level of impairment may affect the ability of subjects to retain their improvement in time.

Dataset Information

Enhancing fluorescent protein photostability through robot-assisted photobleaching.

Publications

Enhancing fluorescent protein photostability through robot-assisted photobleaching.

Similar Datasets

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