Sorting Sounds - A high-throughput microfluidics screening platform for the development of genetically encoded labels for Optoacoustic imaging
Biochemistry
Final Report Abstract
The funded work lay the foundations for a routine use of photoswitching OAI. Allowing the use of genetically encoded reporters which are promising to eventually allow the same ease of use as e.g. the green fluorescent protein (GFP) affords now to fluorescence imaging. Beyond that the funded resources allowed first steps towards genetically encoded photoswitching small molecule and ion sensors which can allow to visualize molecular distributions in whole live animals. Together both technologies are the first step to facilitating whole live animal in vivo imaging with high resolution and genetically encoded tools giving a high-resolution live view into (patho-) physiology. The funded work also contributed to generating the necessary data for a successful application for an ERC Consolidator grant and the formation of an EIC Pathfinder consortium. Moreover, the photoswitching sensors show synergies to photoswitching employed in fluorescence super resolution microscopy and could thus generate additional impact in this direction. The development of the screening technology could produce first demonstrations of the capability. Its full application will require the implementation of the sorting necessary for directed evolution strategies, where the funding resources helped to establish the necessary platform that will help to tackle that obstacle and provide the screening resources needed for the advancement of the new field of “photoswitching Optoacoustics”. The success of the funded work is well documented by press releases from the host institution and general media attention to the publications, and dissemination on open science days (e.g. VHS Munich).
Publications
- Characterization of Reversibly Switchable Fluorescent Proteins in Optoacoustic Imaging. Anal Chem. 2018. 90(17):10527-10535
Vetschera P, Mishra K, Fuenzalida-Werner JP, Chmyrov A, Ntziachristos V, Stiel AC
(See online at https://doi.org/10.1021/acs.analchem.8b02599) - Photocontrollable Proteins for Optoacoustic Imaging. Anal Chem. 2019. 91(9):5470-5477
Mishra K, Fuenzalida-Werner JP, Ntziachristos V, Stiel AC
(See online at https://doi.org/10.1021/acs.analchem.9b01048) - Multiplexed whole animal imaging with reversibly switchable optoacoustic proteins. Science Advances. 2020. 22(9):441-446
Mishra K, Stankevych M, Fuenzalida-Werner JP, Grassmann S, Gujrati V, Klemm U, Buchholz VR, Ntziachristos V, Stiel AC
(See online at https://doi.org/10.1126/sciadv.aaz6293) - In vitro optoacoustic flow cytometry with light scattering referencing. Sci Rep. 2021. 11(1):2181
Seeger M, Stiel AC, Ntziachristos V
(See online at https://doi.org/10.1038/s41598-021-81584-y) - Genetically encoded photo-switchable molecular sensors for optoacoustic and super-resolution imaging. Nat Biotechnol. 2022. 40, 598–605
Mishra K, Fuenzalida-Werner JP, Pennacchietti F, Janowski R, Chmyrov A, Huang Y, Zakian C, Klemm U, Testa I, Niessing D, Ntziachristos V, Stiel AC
(See online at https://doi.org/10.1038/s41587-021-01100-5)
