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An automated liquid jet for fluorescence dosimetry and microsecond radiolytic labeling of proteins.

ABSTRACT: X-ray radiolytic labeling uses broadband X-rays for in situ hydroxyl radical labeling to map protein interactions and conformation. High flux density beams are essential to overcome radical scavengers. However, conventional sample delivery environments, such as capillary flow, limit the use of a fully unattenuated focused broadband beam. An alternative is to use a liquid jet, and we have previously demonstrated that use of this form of sample delivery can increase labeling by tenfold at an unfocused X-ray source. Here we report the first use of a liquid jet for automated inline quantitative fluorescence dosage characterization and sample exposure at a high flux density microfocused synchrotron beamline. Our approach enables exposure times in single-digit microseconds while retaining a high level of side-chain labeling. This development significantly boosts the method's overall effectiveness and efficiency, generates high-quality data, and opens up the arena for high throughput and ultrafast time-resolved in situ hydroxyl radical labeling.

SUBMITTER: Rosi M

PROVIDER: S-EPMC9411504 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature

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An automated liquid jet for fluorescence dosimetry and microsecond radiolytic labeling of proteins.

Rosi Matthew M Russell Brandon B Kristensen Line G LG Farquhar Erik R ER Jain Rohit R Abel Donald D Sullivan Michael M Costello Shawn M SM Dominguez-Martin Maria Agustina MA Chen Yan Y Marqusee Susan S Petzold Christopher J CJ Kerfeld Cheryl A CA DePonte Daniel P DP Farahmand Farid F Gupta Sayan S Ralston Corie Y CY

Communications biology 20220825 1

X-ray radiolytic labeling uses broadband X-rays for in situ hydroxyl radical labeling to map protein interactions and conformation. High flux density beams are essential to overcome radical scavengers. However, conventional sample delivery environments, such as capillary flow, limit the use of a fully unattenuated focused broadband beam. An alternative is to use a liquid jet, and we have previously demonstrated that use of this form of sample delivery can increase labeling by tenfold at an unfoc ...[more]

PMID: 36008591

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Project description:Background Needle-free jet injection (NFJI) systems enable a controlled and targeted delivery of drugs into skin tissue. However, a scarce understanding of their underlying mechanisms has been a major deterrent to the development of an efficient system. Primarily, the lack of a suitable visualization technique that could capture the dynamics of the injected fluid–tissue interaction with a microsecond range temporal resolution has emerged as a main limitation. A conventional needle-free injection system may inject the fluids within a few milliseconds and may need a temporal resolution in the microsecond range for obtaining the required images. However, the presently available imaging techniques for skin tissue visualization fail to achieve these required spatial and temporal resolutions. Previous studies on injected fluid–tissue interaction dynamics were conducted using in vitro media with a stiffness similar to that of skin tissue. However, these media are poor substitutes for real skin tissue, and the need for an imaging technique having ex vivo or in vivo imaging capability has been echoed in the previous reports. Methods A near-infrared imaging technique that utilizes the optical absorption and fluorescence emission of indocyanine green dye, coupled with a tissue clearing technique, was developed for visualizing a NFJI in an ex vivo porcine skin tissue. Results The optimal imaging conditions obtained by considering the optical properties of the developed system and mechanical properties of the cleared ex vivo samples are presented. Crucial information on the dynamic interaction of the injected liquid jet with the ex vivo skin tissue layers and their interfaces could be obtained. Conclusions The reported technique can be instrumental for understanding the injection mechanism and for the development of an efficient transdermal NFJI system as well. Supplementary Information The online version contains supplementary material available at 10.1186/s13036-023-00335-x.

Dataset Information

An automated liquid jet for fluorescence dosimetry and microsecond radiolytic labeling of proteins.

Publications

An automated liquid jet for fluorescence dosimetry and microsecond radiolytic labeling of proteins.

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