Bioengineering unleashed

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.

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Better cell sample preparation with PRIMO,
Better in vitro cell models.

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.

“The fact that we can use the same system to generate multiprotein patterns and even create 3D structures makes it a very powerful setup. “

Project Leader, Jorge Ferreira, i3S, Portugal

PRIMO bioengineering technology

The combination of 3 products specifically designed for prototyping and engineering custom in vitro microenvironments !

Setup PRIMO 2 - PLPP - Leonardo - inverted microscope

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:

  1. SUBSTRATE selection and preparation
  2. PATTERN DESIGN and upload in Leonardo software
  3. UV PROJECTION BY PRIMO for micropatterning, microfabrication, hydrogel polymerization
  4. USING CUSTOM in vitro CONDITIONS for better cell experiments
micropatterning icon
microfabrication icon
hydrogel icon


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.

Unrivalled performance



for cell culture*

*Flat or structured, stiff or soft: slides, coverslips, hydrogels, PDMS, microfluidic chips, etc.

High Resolution


over the entire field of view*

*Approximately 500×300µm,
20x objective



for a full field pattern*

*Approximately 500×300µm,
20x objective, with PLPP Gel



gray levels



depending on
experimental conditions


LLG input

*Excitation light will be reflected by the DMD and projected into the microscope pathway.

Micropatterning applications in cell biology

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).

Whole-cell Cryo-ET:
the challenge of the perfect sample

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
Cells randomly seeded on the surface of an EM grid
Mouse A9 fibroblast cells (labeled with Alexa Fluor 488 Phalloidin visualizing F-actin in green) irregularly distributed on the surface of an EM grid. Credit: Leica Microsystems
Cells micropatterned in the center of the squares formed by the mesh of an EM grid
Fibroblasts adhering on fibrinogen micropatterns precisely positioned in the center of the squares of a carbon EM grid mesh. Alvéole collaboration with Dr K. Grünewald.
HeLa cells grown on micropatterned TEM grid done by Dr Elizabeth Wright with PRIMO
HeLa cells grown on micropatterned TEM grid and stained for live cells with calcein-AM (green) and dead cells (Eth-D1, red). © Dr Elizabeth Wright, University of Wisconsin-Madison.

Advanced cryo-ET cell sample preparation,
by Alvéole and Leica Microsystems

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

Optimized cryo CLEM and cryo ET workflows
thanks to micropatterning

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 !

  • Cells well located and spread on the EM grids,
  • Precise, seamless identification of intra-cellular target components and transfer of image data to different EM solutions
  • Optimal cryo conditions maintained throughout the imaging workflow and sample transfer

Fully Integrated Whole-Cell Cryo-ET Workflow for Higher Throughput

cells micropatterned on EM grid

Controlled adhesion

EM grid vitrification for cell cryo-ET


cryo-CLEM with micropatterning

Correlative Microscopy
→ High success rates

cell FIB milling - facilitated with micropatterning

FIB milling
→ Several cells

icon cryo-electron tomography on micropatterning cell

Electron tomography
→ Several lamellae

icon cryo-ET protein in cellular context

Proteins in cellular context


Time saving 
& independence

Rapidly optimize your experimental conditions by yourself!

High Flexibility

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.

Use your usual substrates

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.

Micropatterning of a wide range of molecules

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.

Epifluorescence capability with PRIMO 2

Epifluorescence capability with PRIMO 2

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.”

Resources and Support

Our team gives you all the tips to successfully conduct your experimental manipulations and go even further!



Application notes

See all resources

For an optimized and personalized control over your experimental conditions, discover complementary PRIMO products.


PDMS Stencil

The multi-well solution

With its multiple wells, the PDMS Stencil reduces the volumes of reagents used and accommodates different experimental conditions on the same substrate.
Discover the PDMS Stencil


The photoactivatable product line

PRIMO is a UV-based photopatterning system with a diverse range of applications including micropatterning and hydrogel structuration. Our product line supports both of these applications with photoactivable reagents. PLPP Classic: enables high-throughput micropatterning of cell culture substrates when illuminated with PRIMO. PLPP Gel: accelerates the speed of your micropatterning experiments and substrate biofunctionalization up to 30 times compared to PLPP Classic. HyPE: enables to flexibly structure hydrogels in 2.5D, while coupled with UV.
Additional information on the PRIMO reagents


New version and new concept

The LEONARDO software has been developed to control the PRIMO system and optimize the usage of its different capabilities. The new LEONARDO 5 is built in modules to improve its ease of use and to save time while performing experiments with PRIMO: micropatterning, EM grids micropatterning, multiwell plates micropatterning, hydrogel structuration, microfabrication, multi-images projection. The new features and modules have been approved and validated by our users.
Discover the features of LEONARDO


New hardware solution with integrated optics

No inverted microscope available for PRIMO2? We are now offering a micropatterning platform with integrated optics. This platform gives you access to the PRIMO2 technology without the need for a dedicated microscope. An all-in-one platform for micropatterning, hydrogel polymerization, and microfabrication. Non-contractual image
Additional information on the micropatterning platform
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