The first multi-protein photopatterning solution
Control several parameters in the cell microenvironment and study their impacts on cell development using an innovative multi-protein photopatterning solution.
For many years, studying the influence of the microenvironment on intracellular and intercellular mechanisms has been essential for research in cell and medical biology. Among the methods for controlling this microenvironment is the rapidly developing process of “micropatterning”, which involves creating protein patterns on which living cells are cultivated. However, current micropatterning techniques are tedious, complex and non-quantitative.
Based on this finding, the scientists at Alvéole developed an innovative multi-protein photopatterning technique to make experimental manipulations easier for researchers.
The PRIMO technique is based on LIMAP* technology (Light Induced Molecular Adsorption of Proteins) and combines a UV illumination system controlled by a dedicated software (named “Leonardo”) and a specific photoactivatable reagent (PLPP). Working together, these two key system components make it possible to generate, in only a few seconds, any multi-protein pattern on standard cell culture substrates.
*“Multiprotein Printing by Light-Induced Molecular Adsorption” Strale P.O. et al, Adv Mater. 2015
Primo was developed to enable you to design and conduct all the micropatterning experiments you can imagine, in 2D and also 3D. Simply select from amongst your computer’s files the pattern you want to use (no size or shape limitations). Primo then projects it on the cell culture substrate and allows you to generate the pattern with the protein of your choice.
We are gradually discovering the extent of the fields of application of this new technology and we are pleased to show you a few examples of some early experiments conducted by our users.
PRIMO allows precise, easy and rapid adjustment of protein micropatterns, either to study the effect of a drug or to imitate physiological conditions. It therefore opens up new possibilities for multiple areas of application, such as stem cell research, and cell-based assays for drug development and predictive toxicology*, to meet major public health challenges.
*Cell-based assays performed using the micropatterning technique constitute an excellent alternative to animal testing.
PRIMO: a UV illumination module. Mounted on a microscope, its optical imaging system projects your chosen pattern onto standard cell culture substrates.
PLPP: an exclusive specific photoactivatable reagent. When coupled with UV action, it enables the pattern to be created on the illuminated area of the substrate. Thus you can precisely apply the protein of your choice.
LEONARDO: the software that provides you with optimal control of the PRIMO module, and facilitates your experimental manipulations.
PRIMO® is a registered trademark of Alvéole SAS. PLPP™ is a trademark of Alvéole SAS.
*Approximately 500x300µm, 20x objective.
Associate Professor at MechanoBiology Institute - National University of Singapore, and Director of Research at CNRS
« My research work consists in understanding how microenvironmental cues influence, guide and shape cell-cell contacts. We are currently particularly interested in determining the role of the biophysical environment in the establishment of apico-basal polarity in mammary gland cells and in liver cells. The use of PRIMO in this context proved absolutely essential since it allowed us to create artificial microniches in 3D where we could control up to 150 combinations of environmental cues.
By printing our protein of choice and microfabricating intricate structures under the microscope, we are gradually uncovering how the geometrical and biophysical parameters drive the elongation of lumen into tubes in the context of hepatic development and diseases.
PRIMO is unique in its capabilities, and for us it is the perfect tool. Submicron-resolution, flexibility and versatility in terms of the proteins you can deal with. It is a very reliable tool to print proteins on surface. »
Director of Laboratory “Adhesion & Inflammation” - INSERM
« The team is studying the recruitment mechanisms of white blood during an immune response. The complex orchestration of white blood cells traffic between blood/lymph systems, lymph nodes and inflammation zones relies on sophisticated but still poorly understood signaling routes that guide cells toward precise targets. Our aim is to develop in vitro experimentation to decipher guiding mechanisms involved in vivo.
PRIMO technology is particularly adapted to design in vitro microdevices patterned with controlled patches of the signaling proteins relevant for white blood cell migration. »
CEA Engineer at the Physics of cytoskeleton & Morphogenesis lab (Cytomorpholab)
« We are working on the generation of 3D cellular microenvironments to reproduce hematopoietic niches. Primo will be used to generate 3D photopolymerised microenvironments and to pattern them to localize different cell populations involved in the hematopoiesis.
We initially chose to use PRIMO because it allows sequential multi patterning and makes gradient patterning possible. It works with a broad range of scale and its design creation is limitless. The technology is reproducible and robust after good practical and highly performant to prototype designs or protein combinations. »
Research Scientist at INM-Leibniz Insitute for New Material
« We are interested in imaging subcellular localization of certain cell surface receptors and check whether they colocalize with focal adhesion complexes. For this purpose we are interested in making different types of patterns of Fibronectin with subcellular dimensions. We decided to use PRIMO because our lab uses photoresponsive ligands to make dynamic biomaterials, so in general this was a useful tool for most of our members. In particular, it suited my requirement of making patterns with different dimensions and features and being able to immobilize any protein of interest onto it. »
Postdoctoral Fellow at UNSW Sydney
« My research project aims at unravelling how a T cell switches from a fast migratory state to a stationary state upon activation. To do so, I perform live cell imaging of T cells migrating inside micro-fabricated channels coated with activating molecules. However, with this approach, I do not control when and where a T cell encounters the activating molecules. The use of PRIMO allows me to print a “migratory zone” and an “activation zone” along micro-channels to visualize this transition. And I also chose to use PRIMO for its capacity to print proteins on 3D surfaces and because it enables me to fine-tune the printing area on a per experiment basis. »
PhD Student at Leiden University Medical Center, Anatomy and Embryology Lab.
The team wanted to have a flexible system which they can use to pattern multiple proteins, gradients on different substrates. Here is Oleh’s opinion on our technology after a few weeks use: « If I should use one word to characterize PRIMO, I would say mind blowing, impressive or unbelievable. »