PRIMO 2, new generation with compact footprint, new optical design allowing epi-fluorescence microscopy and faster performances.
Micropatterning, hydrogel polymerization and microfabrication, all in a single device. Create bespoke in vitro cellular microenvironments and get better cell models for your cell biology experiments or cryo-ET studies.
Studying the influence of the microenvironment on intracellular and intercellular mechanisms has been essential for research in cell biology, for many years now. But in this quest, in vitro cell experiments confront researchers with many challenges, such as:
• the recurring reproducibility issues,
• reliability in term of physiological relevance,
• but also ease of use,
• and efficiency.
This is why our research team has developed PRIMO: a photopatterning system performing micropatterning, microfabrication and hydrogel polymerization to overcome all this issues. We make it evolves to a new generation, PRIMO 2, with compact footprint, optic schematics allowing fluorescence and faster performances.
Project Leader, Jorge Ferreira, i3S, Portugal
The combination of 3 products specifically designed for prototyping and engineering custom in vitro microenvironments !
Developed by Alvéole scientists after the LIMAP* technology (Strale P.O. et al, Adv Mater. 2016), the PRIMO maskless photopatterning system (DMD based) can engineer custom in vitro cell microenvironments through three techniques: micropatterning, hydrogel structuration and microfabrication.
PRIMO workflow for better in vitro cell models:
PRIMO micropatterning system allows to fine-tune cell adhesion to mimic in vivo phenotypes, study cellular mechanisms or isolate cells in reproducible conditions for standardized assays.
PRIMO DMD-based photopatterning system can perform greyscale photolithography on greyscale resists to create complex 3D molds such as ramps, curving wells or microfluidic chips for organ-on-a-chip applications.
As a photopatterning system, PRIMO can also polymerize and photo-scission most commonly used hydrogels for 3D cell culture, spheroid formation or polymerizing permeable membranes in microfluidic chips.
standard
substrates
for cell culture*
*Flat or structured, stiff or soft: slides, coverslips, hydrogels, PDMS, microfluidic chips, etc.
1,2µm
over the entire field of view*
*Approximately 500×300µm,
20x objective
256
gray levels
3
depending on
experimental conditions
LLG input
*Excitation light will be reflected by the DMD and projected into the microscope pathway.
Far from being limited to mechanobiology (known as the in vitro analyze of living cells intra and inter-cellular mechanisms), micropatterning proves to be a powerful asset for different fields of research, such as disease modeling, immunology, toxicology, and more recently for cryo-electon tomography (cryo-ET).
Cryo-ET being still a young microscopy technique, all the steps of its workflow are being intensely reviewed and improved both by academic labs and industrial companies.
By controlling the cell adhesion, spreading and shape, micropatterning overcomes the issues faced at the very first step of the cryo-ET workflow, namely the quality of the cell samples.
Learn more on micropatterning on EM grids
Our main consideration at Alvéole is to continually develop innovative solutions that meet your needs to improve the quality of your cell models for biology and microscopy.
In the case of cryo-ET, we share this philosophy of excellence with Leica Microsystems which led us to join forces and combine our products – PRIMO, Leica’s EM GP2 plunge freezer, THUNDER Imager EM Cryo CLEM and additional cryo CLEM solutions – into a highly efficient workflow!
Learn more on Leica THUNDER Imager EM Cryo CLEM
By fine-tuning the cellular adhesion on EM grids, PRIMO maskless micropatterning system overcomes issues linked to the very first step of the cryo-ET workflow: getting cells amenable for cryo-ET.
Integrate PRIMO micropatterning system as a first step to Leica Microsystems’ Cryo CLEM workflows and make your cryo-ET sample preparation process even more reliable and successful !
Micropatterning
→ Controlled adhesion
Vitrification
Correlative Microscopy
→ High success rates
FIB milling
→ Several cells
Electron tomography
→ Several lamellae
Proteins in cellular context
Rapidly optimize your experimental conditions by yourself!
Download any images you want to structure and / or functionalize your substrate surface!
Use your regular cell culture substrates: flat or microstructured, stiff or soft.
Protein micropatterning: glass coverslips, glass slides, 96 well plates, EM grids, polystyrene, PDMS, polyacrylamide gel (transfer), hydrogels.
Hydrogel structuration: UV-curable hydrogels, PEG Acrylate, Polyacrylamide, Agar, Matrigel.
Microfabrication: UV-curable materials, UV-photoresists.
More than 10 proteins used daily by our users, including:
Fibrinogen-488, Fibrinogen-647, Fibronectin, GFP, Neutravidin-488, Neutravidin-647, PLL-PEG-Biotin, Protein A-647, Streptavidin, as well as primary and secondary antibodies.
Sequential photopatterning of Fibrinogen-A488 in green and Protein A-A647 in red onto PDMS micro-pillars microfabricated with PRIMO.
Epifluorescence microscopy image of 1,5µm dots (spaced by 1,5µm) of ProteinA-488 on PDMS.
Epifluorescence microscopy image of 2µm horizontal lines of ProteinA-488 on glass.
Epifluorescence microscopy image of a gradient of Fibrinogen-A488 on a glass coverslip.
Epi-fluorescence microscopy of a protein micropattern thanks to the optical design of the 2nd generation of PRIMO module.
“My interest is to understand the role of biophysical and topological properties of tissue microenvironments, such as stem cell niches, in modulating cell fate. Thus, the ability to precisely tune and control extracellular cell/organelle shape and geometry in 2D and 3D, is of critical importance. PRIMO has been incredibly useful in this regard!”
“The main interest of the Tardieux’s laboratory and my PhD project is to decipher how forces drive the unique motile and invasive capacities of the single-celled eukaryotic parasite Toxoplasma gondii. I was able to uncover that the parasite glides by coupling polar adhesions and de-adhesion with traction and dragging forces. The PRIMO technique was needed to create composite patterns with a non-adhesive area next to an adhesive one with the crucial request of a sharp demarcation.”
“We are working on the generation of 3D cellular microenvironments to reproduce Hematopoietic Niches. PRIMO will be used to generate 3D photo-polymerized microenvironments and to pattern them to localize different cell populations involved in the hematopoiesis.”
“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.”
“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.”
“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.”
“Protein micropatterning represents an excellent tool to probe the behavior and functions of cellular systems. PRIMO is specially suited for our experiments, in which the cell-substrate interaction needs to be precisely adjusted both throughout the substrates and in time, in order to control the dynamic behaviour of cell monolayers.”
“Our research is at the frontier of soft matter physics and process engineering. More precisely, we develop microfluidic tools to study industrial processes (mixing, flow, drying, filtration, etc.) involving soft matter systems such as polymers or colloids. We use PRIMO to integrate hydrogel membranes in microfluidic devices to mimic ultrafiltration and dialysis processes on the scale of a few nanoliters.”
Our team gives you all the tips to successfully conduct your experimental manipulations and go even further!
Our team will be happy to answer any questions you may have.