Low dosage 3D volume fluorescence microscopy imaging using compressive sensing
This addresses phototoxicity in live embryo imaging, offering a solution for deep imaging modalities like two-photon and light-sheet microscopy, though it appears incremental as it builds on existing low-dosage techniques.
The paper tackles the problem of phototoxicity in 3D fluorescence microscopy by proposing a compressive sensing approach that reconstructs 3D volumes with the same signal-to-noise ratio using less than half the excitation dosage, achieving accurate reconstruction from less than 20% of optical sections.
Fluorescence microscopy has been a significant tool to observe long-term imaging of embryos (in vivo) growth over time. However, cumulative exposure is phototoxic to such sensitive live samples. While techniques like light-sheet fluorescence microscopy (LSFM) allow for reduced exposure, it is not well suited for deep imaging models. Other computational techniques are computationally expensive and often lack restoration quality. To address this challenge, one can use various low-dosage imaging techniques that are developed to achieve the 3D volume reconstruction using a few slices in the axial direction (z-axis); however, they often lack restoration quality. Also, acquiring dense images (with small steps) in the axial direction is computationally expensive. To address this challenge, we present a compressive sensing (CS) based approach to fully reconstruct 3D volumes with the same signal-to-noise ratio (SNR) with less than half of the excitation dosage. We present the theory and experimentally validate the approach. To demonstrate our technique, we capture a 3D volume of the RFP labeled neurons in the zebrafish embryo spinal cord (30um thickness) with the axial sampling of 0.1um using a confocal microscope. From the results, we observe the CS-based approach achieves accurate 3D volume reconstruction from less than 20% of the entire stack optical sections. The developed CS-based methodology in this work can be easily applied to other deep imaging modalities such as two-photon and light-sheet microscopy, where reducing sample photo-toxicity is a critical challenge.