Countstar Castor S1

High-throughput Intelligent 3D Cell Analyzer

With the emergence of organoids as a potential alternative to model human tissue, interest in how these three-dimensional (3D) structures reacts to its environment is growing among research and biopharma-ceutical community. The development of innovative organoid based research or organoid-based therapies demands accurate and significant higher throughput organoid-based experiments that can accurately capture the physiology of these 3D struc-tures. The Castor S1 is a high-throughput image analyzer that enables high-content analysis of thousands of organoids simultaneously. It is equipped with advanced optical technologies with Z-scanning feature, and Artificial Intelligence (AI) based image recognition algorithms, ensuring high quality images and accurate image processing for confident results. Robust software with intuitive user interface provides comprehensive suite of data analysis tools, simplifying data review of large data set. The Countstar Castor S1 is the ideal platform for many high-throughput organoid-based assays, including features to perform tailored applications, such as organoid culture quality, drug-response test, organoid viability, and cytotoxicity analysis for co-culture assay with immune cells.

Core Advantages

• Advanced optical technology allows capturing clear and sharp images in the entire well area ensures accuracy. Peltier-cooled 10MP CMOS high resolution camera, 4.0x and 10.0x high NA objective and a laser-based autofocus system, ensures sharp image acquisition in any situation.
• Dual fluorescence channels (Ex480/30nm – Em535/40nm (green); Ex540/25nm – Em620/60nm (red)) dramatically expand application scope than bright field only platforms.
• Z-stack imaging and projection features to capture all organoid grown on 3D matrigels allowing phenotyping each organoid grown on matrigels.
• AI-based, highly customisable image analysis algorithms improve image processing accuracy.
• Broad consumable compatibility with common 6-384 well plates, petri dish, culture flask, and slide provides significant time saving and optimum flexibility.
• Easy to use, and complaint-ready software meets FDA 21 CFR Part 11 requirements.
• Possible integration with various automation platforms further increases through-put.

Application Detailes

Applications Overview

The Countstar Castor S1 is the ideal platform for many high-throughput organoid-based assays, including features to perform tailored applications, such as organoid culture quality, drug-response test, organoid viability, and cytotoxicity analysis for co-culture assay with immune cells.

Selectd Applications, Results, and Graphics

I、Automatic analysis of quality of organoid culture

The Countstar S1 automatically tracks each individual organoid in the entire culture well plate throughout the organoid culture duration. Its Z-stacking feature and AI powered algorithm enables comprehensive phenotype analysis of each organoid.

II、Growth monitoring of cancer-derived organoids

Countstar S1’s innovative optical design allows capturing bright and clear images of the entire well area in a culture plate. Its AI powered algorithm and automated data analysis ensures accurate and high throughput measurement of organoids morphology during their growth time in each individual cells in the culture plate.

(a) Plate view with thumbnail images of hepatic cholangiocarcinoma organoids over 7 days; (b) Enlarged, processed image of the selected well D3 with identified organoids colored differently, according to their phenotypes; (c) Time course plot of average organoids diameter development of the selected well.

III、Label-free organoid drug response test

Organoids with diverse morphologies in a complex sample can be accurately identified and char-acterized using Z-stacking feature and AI-based image analysis algorithms.
Organoids derived from cisplatin-resistant primary cholangiocarcinoma tissues were cultured for 5 days, then treated with either cisplatin or gemcitabine for 3 days in replicates. Samples were then analyzed in a Countstar Castor S1. Gemcitabine showed strong toxicity to the organoids, while cisplatin didn’t stop the growth of the organoids.

Live organoids are labeled in green. Dead, degrading organoids are labeled in red.

IV、Organoid Fluorescence Viability Analysis

With bright field imaging capability and dual fluorescence channels, cell viability in organoids and tumor spheroids can be measured using viability staining reagents (e.g. Acridin Orange (AO) / Propidium Iodide (PI). Fluorescence imaging in a Z-axis scan is carried out to obtain the titer, distribution, and spatial localization of dead and live cells. This is essential in evaluating the tissue penetration effect of drugs.

Live cells labeled red. Dead cells labled green

V、Cytoxicity Analysis for Tumor Organoid-Based Immuno-Oncology (IO) Co-Culture Assay

Patient-derived organoids (PDO) are a powerful and effective model for drug screening and patient response prediction to treatment regimens, especially with immune cells co-culture. The Countstar Castor S1 enables high-throughput imaging and effective data processing, accelerating the identification of an optimal, individualized treatment for the patients.

Proliferating hepatocellular carcinoma (HCC) organoids are labeled in green (CFSE); Dead and degrading HCC organoids and NK-92MI cells are labeled in red (EthD-1)