SUN Bioscience uses ImageXpress® Micro Confocal High-Content Imaging System to image high-throughput organoid platform
TEAM MEMBERS
Maria Clapés Cabrer
SUN bioscience
PRODUCTS USED
ImageXpress Micro Confocal
IN Carta Image Analysis Software
The Challenge
One of the most significant challenges facing researchers using complex 3-dimensional (3D) microtissues is collecting accurate and reliable high-throughput images and data. SUN bioscience’s innovations in the field of organoid and 3D cell culture aim to standardize organoid culture through their proprietary high-throughput microwell platform, Gri3D®.
Gri3D microwell plates allow the generation of multiple organoids of similar sizes in the same focal plane, which requires less imaging time and less image storage space. However, to take full advantage of this, a high-throughput, high-content 3D imaging system is required to enable scientists to move towards higher-throughput organoid assays. Such a solution is important not only for SUN bioscience’s customers, but also for the company itself as they develop their plates, generate proof-of-concept studies and demonstrate what is possible using their solutions.
The Solution
SUN bioscience discovered that the ImageXpress Micro Confocal High-Content Imaging System from Molecular Devices integrates well with their Gri3D to provide a platform that enables high-throughput, high-content organoid imaging, maximizes datapoint collection and reduces imaging time. The ImageXpress Micro Confocal significantly expedites the imaging process, enabling in situ brightfield and fluorescence imaging at a range of magnifications (low at 4x, mid at 10-20x, and high at 40x) directly on the plate. This not only accelerates the assays, but also facilitates quicker and more efficient imaging and subsequent analyses. Brightfield imaging over time provides information on the dynamics of organoid development. This label-free imaging approach coupled with AI-enabled IN CARTA(r) image analysis software is used by SUN bioscience to analyze and characterise each organoid.
Products Used
The ImageXpress Micro Confocal system is a high-content solution that can switch between widefield and confocal imaging of fixed and live cells. It can capture high quality images of whole organisms, thick tissues, 2D and 3D models, and cellular or intracellular events. The spinning disk confocal and sCMOS camera enable imaging of fast and rare events like cardiac cell beating and stem cell differentiation. With MetaXpress software and flexible options like water immersion objectives to choose from, the system enables many confocal imaging applications from 3D assay development to screening.
The Results
So far, SUN bioscience has run several assays on the ImageXpress system, including monitoring individual organoids in brightfield, proof-of-concept screens on gastrointestinal organoids and pancreatic cancer tumoroids, as well as modelling inflammatory bowel disease – specifically the effects of a cocktail of cytokines on the viability and cytoskeleton of human rectal organoids.
Response of pancreatic cancer patient-derived organoids exposed to anti-cancer compounds for 72 hours. A. TL images before (0 h) and after (72 h) exposure and maximum projection images of organoids after Live/Dead assay at 72h. Green: Calcein AM, live; red: EthD-1, dead. B. Ethidium homodimer-1 (EthD-1) to Calcein AM intensity ratio. C. Grey levels non-normality factor (GLNN) from TL images. Error bars show standard deviation. Each dot represents an organoid. One-way ANOVA Dunnett’s multiple comparisons, **P < 0.01, P**** < 0.0001, ns: non-significant. Microwell size: 500 µm
High magnification confocal images of human rectal organoids exposed to pro-inflammatory cytokines. 4oX single plane images of F-actin (magenta) and DAPI (blue). Arrows indicate regions of interest where F-actin signal shows a defined structure in both control and infliximab conditions. Delocalization of F-actin is highlighted in the inflammatory cocktail condition.