Periodic Reporting for period 1 - SWOPT (Breaking the penetration limit of microscopy – Photoswitching Optoacoustics)
Okres sprawozdawczy: 2022-09-01 do 2023-08-31
SWOPT aims to solve one of the great challenges in visualizing biology: break through the penetration limits of optical microscopy and image cells and their function in vivo at depths of several millimeters to centimeters, while retaining high resolution and sensitivity of single cells. The ability to visualize few cells in a live organism is important because many biological phenomena, especially in the immune system, the onset of tumors, or fundamental developmental biology, rely on a small number of cells.
SWOPT will achieve this by combining the principles of photoswitching with optoacoustic imaging. Optoacoustic imaging is a method that relies on reading out ultrasound signal generated by light. It already has the power to deliver a combination of higher penetration depth, higher resolution, and larger fields of view than other imaging technologies. However, for many research questions, optoacoustics needs tools like genetically encoded reporters and sensors. Photoswitchable label proteins can help here. These proteins can change their state upon illumination, like a tiny switch that changes the state of the protein from ON to OFF and with that, the signal it generates. In optoacoustics, we use the light switchable signal to make the label blink, which enables the visualization of small numbers of cells against a strong background of other signals, which shows a constant signal. One can imagine the effect, like the blinking of a lighthouse in a stormy dark night at sea.
SWOPT will enable examination of whole tissues in vivo with the same ease, flexibility and, eventually, abundance of tools, paralleling fluorescence microscopy, thus bringing research and understanding of living organisms to the next level. As an affordable imaging technology, SWOPT aspires to become routine in life science and bio-medical research.
The overall objective of SWOPT is to create a technology based on optoacoustic imaging and photoswitching. To this aim, a new imaging instrumentation will be developed, and SWOPT-tailored contrast agents (protein-based and synthetic) will be created. SWOPT’s abilities will be benchmarked by visualizing cells and their anatomical and chemical environment in vivo in whole tumors.
SWOPT will achieve this by combining the principles of photoswitching with optoacoustic imaging. Optoacoustic imaging is a method that relies on reading out ultrasound signal generated by light. It already has the power to deliver a combination of higher penetration depth, higher resolution, and larger fields of view than other imaging technologies. However, for many research questions, optoacoustics needs tools like genetically encoded reporters and sensors. Photoswitchable label proteins can help here. These proteins can change their state upon illumination, like a tiny switch that changes the state of the protein from ON to OFF and with that, the signal it generates. In optoacoustics, we use the light switchable signal to make the label blink, which enables the visualization of small numbers of cells against a strong background of other signals, which shows a constant signal. One can imagine the effect, like the blinking of a lighthouse in a stormy dark night at sea.
SWOPT will enable examination of whole tissues in vivo with the same ease, flexibility and, eventually, abundance of tools, paralleling fluorescence microscopy, thus bringing research and understanding of living organisms to the next level. As an affordable imaging technology, SWOPT aspires to become routine in life science and bio-medical research.
The overall objective of SWOPT is to create a technology based on optoacoustic imaging and photoswitching. To this aim, a new imaging instrumentation will be developed, and SWOPT-tailored contrast agents (protein-based and synthetic) will be created. SWOPT’s abilities will be benchmarked by visualizing cells and their anatomical and chemical environment in vivo in whole tumors.
Since the start of the project, SWOPT has made significant progress on the design of the new instrumentation, algorithm and the photoswitchable reporters.
Novel instrument development requires a stable foundation from which to embark. For SWOPT we set up a dedicated open architecture of the raster scanning optoacoustic mesoscopy well established in the work of partner Ntziachristos and marketed by partner iThera medical. This open system we currently use to test the illumination and ultrasound detection capabilities, the interplay with the photoswitching labels and acquire the first testing date for algorithm development.
A crucial step to acquire a three-dimensional image of the different photoswitchable protein labeled cells in tissue is to develop suitable mathematical algorithms. Towards this end, we have already specified the mathematical model, which includes the successive steps from light excitation to ultrasound measurements in the context of photoswitching.
Regarding the labels the chemistry partner has several lead candidate compounds that show promising photoswitching behavior. Those compounds are now tested for their optoacoustic signal generation.
For the genetically encoded sensor engineering the main target candidates in terms of kidney cancer relevant metabolites has been identified and first constructs for a fluorescent sensor have been established. This fluorescent sensor serves as a foundation to embark on the development of the optoacoustic photoswitching sensor.
Novel instrument development requires a stable foundation from which to embark. For SWOPT we set up a dedicated open architecture of the raster scanning optoacoustic mesoscopy well established in the work of partner Ntziachristos and marketed by partner iThera medical. This open system we currently use to test the illumination and ultrasound detection capabilities, the interplay with the photoswitching labels and acquire the first testing date for algorithm development.
A crucial step to acquire a three-dimensional image of the different photoswitchable protein labeled cells in tissue is to develop suitable mathematical algorithms. Towards this end, we have already specified the mathematical model, which includes the successive steps from light excitation to ultrasound measurements in the context of photoswitching.
Regarding the labels the chemistry partner has several lead candidate compounds that show promising photoswitching behavior. Those compounds are now tested for their optoacoustic signal generation.
For the genetically encoded sensor engineering the main target candidates in terms of kidney cancer relevant metabolites has been identified and first constructs for a fluorescent sensor have been established. This fluorescent sensor serves as a foundation to embark on the development of the optoacoustic photoswitching sensor.
The development of the SWOPT technology will set a new level of imaging whole opaque organism in vivo with direct impact on biomedical research. We aim to primarily target SWOPT towards the pre-clinical research market, for which our proof-of-concept experiments in cells and in mice become crucial. Upon validation and exploitation, SWOPT may be used in future studies to visualize cells, their functionality, and their chemical surrounding in context with the complex tissue anatomy. This will contribute to understanding cell specific metabolic pathways that underlie pathological mechanisms of a wide range of diseases.
Additionally, the use of SWOPT is not restricted to live animal tissues, but also expands, with further research, to the emerging large tissue organoids market or explanted organs mimicking disease states. In this sense, SWOPT aspires to become a game-changer for biomedical research, contributing to the development of new disease models and therapeutics.
Additionally, the use of SWOPT is not restricted to live animal tissues, but also expands, with further research, to the emerging large tissue organoids market or explanted organs mimicking disease states. In this sense, SWOPT aspires to become a game-changer for biomedical research, contributing to the development of new disease models and therapeutics.