Microfluidics in life science:

more than just single cell

When using bulk methods, insights into the heterogenous nature of cell populations is lost, this is why microfluidics has changed the way that scientists carry out their single cell protocols. Microfluidics enables truly single cell workflows, without adding additional time, effort, or expertise in many cases. Dolomite Bio was launched to bridge the gap between biologists and microfluidics, allowing researchers to complete single cell RNA-sequencing without the need for prior microfluidic knowledge or experience. The Nadia Instrument, Nadia Innovate and Nadia Go systems give users the option to perform pre-optimized protocols or develop their own scRNA Seq protocols.

Microfluidics can revolutionize not only single cell research, but it can also be used in a wide range of applications. With that in mind, Dolomite Bio has become Do Bio. In our mission to make microfluidics accessible we have utilized our decades of microfluidics experience to develop flexible, open systems, which can process a wide variety of biologicals, from cells, to mRNA, to protein and lipids. Researchers can develop cutting edge protocols and applications for life-science and pharmaceutical research including droplet genomics, hydrogel formation and customization. You’re not limited to just single cell research.

Why use microfluidics?

You may be surprised to hear that microfluidics is already used in everyday laboratory instrumentation: FACs machines, bio-analyzers and mass spectrometry. Microfluidic devices can precisely manipulate small volumes of fluids, producing large numbers of reaction vessels in a short period time, giving researchers the ability to perform multiple experiments simultaneously. Saving you time and money!

Figure 1: Microfluidics provides precise control of the mixing of liquids and gases at a microlitre scale to produce a) droplets and b) flow focusing.

Our main application areas

The Do Bio team contains engineers and scientists who are experts in biomolecules. We have developed multiple products (microfluidic devices and reagent kits) for life scientists that aid with the development of novel methodologies, without compromising on scalability and reproducibility.

1. Droplet Genomics

– insight into single cell omics

Droplet genomics allows the genetic material of individual/multiple cells to be captured in picolitre sized reaction vessels (droplets) in a high throughput and reproducible manner, giving higher resolution over traditional batch methods. Using microfluidics to encapsulate cells can reveal high quality information regarding cellular heterogeneity, leading to the identification of novel sub-cellular populations and gene expression patterns. This information can be used to understand disease development, immunity, and tumor evolution.

Do Bio is known for its innovative products within the droplet genomics research field. The RNAdia 2.0 reagent kit was launched in November 2022 and is used to perform single cell RNA sequencing on cell populations. Single cells/nuclei are encapsulated alongside barcoded mRNA capture beads within a droplet. Cells are lysed, mRNA is reverse transcribed and cDNA libraries are prepared for sequencing using Illumina technologies.

This easy-to-use protocol has been pre-optimized by our in-house scientists to produce consistent, monodispersed droplets when encapsulating a range of biologicals using the Nadia platforms. Samples can be run in parallel, with the option to run up to 8 samples at once. The Nadia instrument offers unique stirring technology within plastic, single use microfluidic chips. This patented technology prevents clumping of cells and beads, maintaining a low doublet rate, without damaging fragile cells.

2. Hydrogels – encapsulate cells in biologically relevant environments.

Made from naturally occurring or synthetic hydrophilic polymers, hydrogels are semi-solid scaffolds that allow the diffusion of water and nutrients. Encapsulating cells within hydrogels can be extremely beneficial for Life Science research, as cells remain viable for extended periods of time. Often proliferating throughout the scaffold, while maintaining cell singularity, making them a versatile tool for understanding antibody discovery and organoid growth.

Depending on the microfluidic set up, biologicals can be encapsulated alone or co-encapsulated with different materials. As a result, hydrogels can be utilized in a range of applications: secretome studies, cell-cell interactions, and high molecular weight preparation.

Temperature control is an important factor to consider when making hydrogels, typically the polymer e.g. agarose used to form the hydrogels requires a high temperature to remain molten and flow through a microfluidic chip.

With this in mind, the Do Bio engineers designed the Nadia platforms to contain an internal temperature control unit capable of heating and maintaining a chip at 40˚c. The internal stirrers are utilized again to prevent the polymerisation of the polymer within the chip reservoir minimising the chance of chip blockage.

The nadAROSE kit was also developed for use on the Nadia instrument. This kit contains everything needed to encapsulate biologicals within an agarose hydrogel. Allowing the encapsulation of thousands of cells within a single run, making this kit extremely cost effective. Following encapsulation, the hydrogels produced are compatible with FACs sorting machines which may be required for many downstream applications. Don’t worry if your biological is rare, you are able to digest the scaffold and release your biological for further downstream testing if needed.

But what if I want to encapsulate a biological in another type of polymer? The Nadia Innovate and Nadia Go allow you to create and customize protocols regardless of your level of microfluidic expertise. Learn more about these systems in the next section.

3. Customization – Open systems for unrivalled versatility

To make ground-breaking discoveries the flexibility and versatility that most commercially available microfluidic systems lack is needed. These systems are designed to be used with common cell types and pre—defined protocols. Here at Do Bio we understand the importance of customizing your workflow to suit the needs of your reagents and biologicals. We developed the Nadia Innovate and Nadia Go as open systems that allow you to adapt critical microfluidic parameters using easy-to-use software: time, pressure, stirrer speed and temperature. The effects of changes can be seen in real time using a high-powered microscope that gives visualization of the chip junction.

Temperature is an extremely important factor when considering biological reactions, requirements can change depending on the experiment. Too high a temperature: enzymes will denature. Too low temperatures can cause polymerisation at the wrong moment. Using the Nadia Innovate or Go, temperature can be adjusted and maintained to meet your experimental needs.

Similarly with time, different applications may require different incubation times. Or you may have a limited quantity of sample so want to shorten the length of one of our standardized protocols. With our open systems, you decide.

Stirrers ensure your biologicals remain in suspension. All Nadia chips contain two stirrers, which can be controlled independently. Changes can be made depending on the density and fragility of your samples.

The pressure delivery to sample and oil lines can alter the size of droplets/particles made using microfluidic devices. Droplets of different sizes, which can be produced with pressure fluctuate during a run, can have different chemical properties, and ultimately give different results even with the same experimental conditions. Using the Nadia Innovate and Go, users can optimize their droplets to meet the needs of their chosen application, create a custom protocol and run this protocol numerous times, achieving reproducible, monodisperse droplets for downstream testing. If using the Nadia Innovate, users have the option to transfer their fully developed protocol to the Nadia instrument and run up to 8 samples in parallel. Making this the ideal system for researchers interesting in optimization and production.

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