White Papers and Tech Notes

Intact Spheroid and Organoid Analysis, Dr. John O'Rourke

BennuBio at World Organoid Research Day 2024

BennuBio Whitepaper


Ditch The Dissociation: A Simplified Approach
for Rapid High Throughput Multiplexed Analysis
of Intact 3D Multicellular Spheroids using the
Velocyt Cytometer

Traditional flow cytometry is often used for the analysis of spheroids but requires disruption of spheroid structure prior to analysis. As a result, the characteristics of individual cells within the spheroid may be altered, and information on spatial organization is lost. Also, the spheroid dissociation workflow is time consuming and requires optimization, the needed washes and centrifugation steps often results in cell loss, especially of non-viable or apoptotic cells, and the enzymatic and manual dissociation methods can damage cells. In this application note, we demonstrate the use of the Velocyt® cytometer for the spheroid workflow using staurosporine treatment as a model drug compound. In this case study, we illustrate Bennubio’s simplified spheroid solution approach (S3 ) to generate and treat large numbers of uniform spheroids, a simplified sample preparation workflow and the rapid analysis of thousands of intact spheroids, generating flow cytometry, fluorescence imaging and morphological data from each spheroid. 

Download the full App Note


Simplified & High Throughput Analysis of Intact 3D Models Using Large Particle Cytometry

The advantages of growing organoids and iPSC aggregates in suspension cultures are manifold, crucial for various applications, scalability, and adaptability to bioreactors. Nevertheless, the analysis of 3D suspension cultures entails tedious workflows, including the transfer of single particles into multiwell plates for imaging, paraffin embedding of particles for histology, or the dissociation of particles into single cells for flow cytometry analysis. To overcome these challenges, we used the Velocyt cytometer by BennuBio to analyze intact 3D models from suspension and static cultures. The Velocyt uses parallel stream acoustic flow technology along with camera-based optics to combine the imaging power of microscopy with the speed and ease of flow cytometry.

Download the poster