Low-cost optofluidic add-on enables rapid selective plane illumination microscopy of C. elegans with a conventional wide-field microscope

Abstract

. Significance : Selective plane illumination microscopy (SPIM) is an emerging fluorescent imaging technique suitable for noninvasive volumetric imaging of C. elegans . These promising microscopy systems, however, are scarce in academic and research institutions due to their high cost and technical complexities. Simple and low-cost solutions that enable conversion of commonplace wide-field microscopes to rapid SPIM platforms promote widespread adoption of SPIM by biologist for studying neuronal expressions of C. elegans . Aim : We sought to develop a simple and low-cost optofluidic add-on device that enables rapid and immobilization-free volumetric SPIM imaging of C. elegans with conventional fluorescent microscopes. Approach : A polydimethylsiloxane (PDMS)-based device with integrated optical and fluidic elements was developed as a low-cost and miniaturized SPIM add-on for the conventional wide-field microscope. The developed optofluidic chip contained an integrated PDMS cylindrical lens for on-chip generation of the light-sheet across a microchannel. Cross-sectional SPIM images of C. elegans were continuously acquired by the native objective of microscope as worms flowed in an L-shape microchannel and through the light sheet. Results : On-chip SPIM imaging of C. elegans strains demonstrated possibility of visualizing the entire neuronal system in few seconds at single-neuron resolution, with high contrast and without worm immobilization. Volumetric visualization of neuronal system from the acquired cross-sectional two-dimensional images is also demonstrated, enabling the standard microscope to acquire three-dimensional fluorescent images of C. elegans . The full-width at half-maximum width of the point spread function was measured as 1.1 and in the lateral and axial directions, respectively. Conclusion : The developed low-cost optofluidic device is capable of continuous SPIM imaging of C. elegans model organism with a conventional fluorescent microscope, at high speed, and with single neuron resolution.

High-throughput light sheet imaging of adult and larval C. elegans Parkinson's disease model using a low-cost optofluidic device and a fluorescent microscope

“…[24][25][26] In a recent study of our group, we reported on developing a low-cost light sheet add-on that can be operated with conventional microscopes to solve the technical complexity and cost of conventional light sheet systems while making the imaging system accessible to researchers. 21 We used an opto-uidic chip with a low-cost PDMS-integrated lens for light sheet formation with lateral and axial resolutions of 1.1 mm and 2.4 mm full-width at half-maximum (FWHM), respectively. While in this preliminary work, we showed the ability to image an entire worm; the system suffered from key limitations such as the inability to continuously image worms' populations, lack of adaptability to image worms at different developmental stages, and lack of imaging consistency to enable quantitative uorescent expression studies.…”

“…This paper reports on optimizing our preliminary technology to overcome its limitations. The main objective is to increase the throughput of Behrouzi's study 21 while keeping the cost of fabrication similar. We utilize a similar cost-effective method for the fabrication of our light-sheet optofluidic system.…”