Our overarching strategy is to precisely 3D structure of the cellular environment our single cells or few cells to elicit a controlled development of the cell-cell contact.
To this end we devised a new method to create microniches. They comprise of a perforated membrane created by capillary filling. The membrane serves as a confining device. It is made of a UV curable polymer indexed matched with cell culture medium to allow precise imaging (MYPOLYMER). We then use on-demand photolithography (PRIMO by Alveole) to pattern the polymeric scaffold in 3D with various calss of protein as well as ECM hydrogels.
WE recently demonstrated that this approach allows creating up to 150 different types of environments (Stoecklin et al Advanced Biosystems, 2018).
The niches are scalable from single cell to organoid size. We call our system Membwell. It is available on demand if you want to try it for your own research.
Single objective SPIM
(Coll R. galland, V.Studer, J.B Sibarita)
The individual microniches described above can be flanked with individual micro mirror. It allows the use of a single objective to illuminate the sample from the side and detect the florescence signal. Using a custom made beam steering system, we used this above property to perform single molecule detection and super resolution imaging inside single cells up to tens of micron above the cover slip (Galland et al, Nature Methods 2015, Sing et al Biophys J 2017).
The system is also scalable and offers the highest photon collection sensitivity allowing the live imaging o very delicate cells (stem or hepatocytes).