Now you can you study cell behavior under confinement

AgarSqueezer is a microscope slide chamber equipped with a molded agar-based compression system. It is used to assess cell response to short and long-term mechanical confinement within a physiological rigidity range.

AgarSqueezer is very helpful if you want to analyze how your cells will react if you squeeze them for a prolonged period. Or if you want to study how mechanical confinement affects drug cell resistance. And if you want to perform immunostaining in situ.

Agarsqueezer has been successfully used with:

• Human cells: primary T-lymphocytes, TF1 & ML2 leukemic cells, HS27A fibroblasts, MCF10A breast cells, MDA-MB-231 breast cancer cells, U-2 OS osteosarcoma cells, PC-3 & DU 145 prostate cancer cells, HT29 & HCT116 colorectal adenocarcinoma cells, HT1080 fibrosarcoma cells and megakaryocytes
• Murine: osteocyte-like cells MLO-Y4, primary muscle cells & primary dendritic cells
• Plant cells: from Arabidopsis roots
• 3D cell objects: mice gastruloïds

How does it work?

8 Features we like

• Tunable stiffness in a physiological range [1-150] kPa: Use of agarose as a cheap and biocompatible polymer
• Open access to the reservoir: Possibility to add drugs, and reagents. Easy medium renewal
• Autoclavable & reusable systems
• Compatibility with multiple microscopy techniques: Confocal, spinning, super-resolution. Open access for microscope objectives. Use of optical glass coverslip to make cells grow
• Easy to recover coverslip with cells for subsequent molecular analysis: FACS, qPCR, Western-Blot, Immunofluorescence (possible in situ)
• Long-term analysis of the cell adaptation to confinement: Up to several days, for time-lapse studies
• Study of the specific impact of mechanical loads on the biology of cells: Gas permeability of the system allows to get rid of the hypoxia conditions
• Easy to assemble and disassemble the system.

AgarSqueezer was developed by Audrey Prunet, Gilles Simon, Hélène Delanoë-Ayari, Véronique Maguer-Satta and Charlotte Rivière.

Charlotte and Véronique, can you please tell us how and why you decided to transfer your invention?

Charlotte Rivière: “In the beginning, we wanted to analyze the influence of both stiffness and confinement on cells, mimicking highly confined situations such as fibrosis or cancer.”

Véronique Maguer-Satta: “On our side, our team was looking for a way to analyze the effect of long-term confinement with the ability to add a drug at any time. We all wanted to find a device that would also meet the requirements of subsequent classical molecular analysis (easy cell culturing and cells recov​ery, qPCR, western-blot, in situ immunostaining) as well as biophysical image-based analysis (high-resolution microscopy and video-microscopy). But we could not find any.”

Charlotte Rivière: “So all together, we decided to collaborate on the perfect set-up. We rapidly identified agarose as an interesting material to get medium and oxygen renewal, with no drug adsorption. We built several prototypes, improving precise control of the confinement, avoiding destruction of the gels and leakage of the culture medium! And then we came up with the Softconfiner device, that we published in Lab on a chip and decided to transfer with Idylle under the name of AgarSqueezer.

Today, we believe that the AgarSqueezer could be of interest for many researchers willing to better understand how mechanics can regulate cell behavior. We hope that researchers from different communities will also find it a useful tool!”