High-resolution extended-depth phase-engineered objectives to accelerate spatial 'omics R&D through computational optics

1 Summary 2 This SBIR Phase II project is focused on the development and validation of groundbreaking engineered point 3 spread function (ePSF) microscope objective lenses (OL) that will empower the next generation of spatial omics 4 investigation. The proposed line of ePSF-OL addresses the critical need in spatial omics for rapid high-content 5 imaging to fully capture cellular identity. Leveraging the power of point spread function engineering to extend the 6 depth of field and encode 3D information directly into the optical response of the imaging system enables 7 volumetric imaging with many fewer axial steps, speeding image acquisition, reducing dataset sizes, reducing 8 phototoxicity and photodamage, and shortening the cycle time to observe live cell dynamics. All these benefits 9 are achieved in the same form factor as standard microscope objective lenses, without reducing the 10 numerical aperture (NA) of the system, thus preserving high resolution data capture. 11 Despite extraordinary advances in optical microscopy, state-of-the-art solutions have been slow to market, 12 lacking in flexibility and ease of access. The ePSF-OL are based on an integrated design of the objective lens, 13 phase modulation, and recovery software. Specifically, two types of ePSF-OL will be developed – Deep Focus 14 (DF) ePSF-OL extends the depth of field for volumetric projection imaging, and Single Helix (SH) ePSF-OL 15 extends the depth of field and encodes 3D spatial information for full 3D imaging. Recovery software will target 16 the needs of spatial omics – localization and counting of biomolecules, and efficient high-resolution restoration 17 of spatial context from fewer axial slices. The ePSF-OL will be designed to achieve the same magnification, NA, 18 and field number as popular objective lenses used in spatial omics assays, thus achieving plug-n-play upgrade 19 compatibility in commercial closed-box, research, or lab-built optical microscopes. 20 This project targets commercialization of cost-effective ePSF-OL offered with novel phase modulation masks 21 that extend the depth of field up to 10 times as compared to standard objective lenses. These commercial- 22 ready prototypes will include a robust opto-mechanical design with excellent performance, supported by GPU 23 accelerated and AI-driven software. Tests of the ePSF-OL and software in advanced spatial omics assays at 24 partner labs will validate end-user acceptance and provide valuable feedback toward commercialization. 25 The implications in spatial omics are far-reaching. For instance, the ePSF-OL will benefit the study of 26 neurodegenerative disease, owing to their ability to simultaneously track proteins and capture direct quantitative 27 readout of transcriptional induction. The ePSF-OL technology will also greatly accelerate error robust 28 transcriptional sequencing, high content proteomics, and the study of epigenomic markers. 29 Double Helix Optics, a startup with exclusive rights to the ePSF technology from U of Colorado, is headquartered 30 in the BioFrontiers Institute in Boulder, CO, and optimally positioned to successfully bring this product to market.