This article details a method for imaging tumor spheroids using optical coherence tomography (OCT). The process allows for the assessment of spheroid morphology and viability over time.
Begin with a multi-well plate containing tumor spheroids, a three-dimensional cancer cell aggregate.
Place it on an optical coherence tomography, or OCT imaging stage, ensuring it is stabilized for accurate imaging.
Adjust the lens for optimal light focus, set the scanning parameters, and initiate image capture over time.
As the light interacts with the cells, it reflects at varying intensities depending on the cellular structures and depths.
OCT captures these reflected light intensities and constructs high-resolution, two-dimensional cross-sectional images at different planes.
Move the plate to acquire the images of tumor spheroids from all wells.
Using software, align these images and process them to create a three-dimensional spheroid model, revealing its morphological characteristics, including size, shape, and structure.
Over time, cells in the spheroid die, and their damaged structures increase light scattering, reducing the reflected light intensity.
This differentiates viable cells from non-viable ones, enabling the monitoring of physiological conditions.
On the day of the tumor spheroids imaging, remove the multi-well plate from the incubator. Transfer it under the OCT imaging system and place it on the plate adapter. To adjust the height of the plate, move it along the z direction of the translation stage. In the custom imaging software, set a desired OCT scanning range to cover the whole tumor spheroid, depending on its developmental stages, and click Save parameters to save the setting.
Then, show the optimized xz and yz OCT previews of tumor spheroids. Acquire 3D OCT images of tumor spheroids one by one for all the wells of the plate containing spheroids. To view the preview image, click the Preview button. And to acquire the OCT image, click the acquire button. Record the overall stage moving process of OCT data acquisition.
- To ensure the optimal image quality for all the tumor spheroids, the plate adapter need to be tuned accurately, and the medium volume need to be the same.
- Use a custom C++ processing code to process 3D OCT data sets of tumor spheroids to generate OCT structural images. Use 2D OCT images in 3 cross-sectional xy, xz, and yz planes across the centroid of the spheroid to generate the collage of spheroid images.
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