Away from the beaten tracks: customize your protocols with the Nadia Innovate and make single cell research work for you
The use of high-throughput single cell techniques, such as single cell RNA-Seq (scRNA-Seq) has expanded exponentially in recent years in research areas as diverse as cancer research, reproductive health, immunology and neurology.
With single cell research, scientists can study gene expression patterns in their chosen samples to identify cell types, cell states and cell heterogeneity. Several commercial platforms are available to implement high-throughput single cell sequencing. Some of them, such as the Nadia Innovate, are based on microfluidics techniques and involve the capture of single cells in microfluidic droplets along with barcoded mRNA capture beads. A drawback of many of these instruments, however, is that their use tends to focus on commonly used cell types, i.e. mammalian cell types, with predefined consumables and protocols.
Narrowly defined procedures are not always compatible with less typical cell types due to variability in cell size, buffer composition and more. This highlights the requirement for adjustments of the single cell capture step and the need for versatile technology to successfully apply the capabilities of single cell research to non-mammalian organisms.
Dolomite Bio’s Nadia Innovate fulfills the need for flexibility within single cell research by enabling open protocol development. It is also worth noting that scientists working on canonical cell types may also wish to develop new protocols to fit with their specific experimental needs or reduce cost of the protocols performed. In this case flexible technology would be of high value to the research team.
So, what are commonly experienced problems or shortcomings in single cell research and how can the Nadia Innovate help resolve them?
Problem: I work on plant protoplasts, they are not compatible with standard buffers such as PBS.
Solution: Plant protoplasts typically require buffers that are more viscous than PBS and this will influence droplet formation. On the Nadia Innovate, you can change pressure parameters to achieve stable droplet formation with your buffer of choice.
Problem: I work on cardiomyocytes (10 x 100 µm), I am concerned that my cells are too big to fit in the droplets.
Solution: The Nadia platform is particularly suited for samples with larger cell sizes. Further, changing pressure parameters on the Nadia Innovate can also be used to alter the droplet diameter to suit a particular cell size.
Problem: I want to encapsulate cells/particles that do not remain in suspension for very long. How do I prevent them from settling during a run?
Solution: The Nadia Innovate is equipped with proprietary stirrer technology which allows stirring of both cells and particles (e.g., mRNA capture beads) during a run. The user can stir cells and beads at different speeds from each other, between 0 and 300 rpm.
Problem: A typical scRNA-Seq Nadia run requires 75,000 cells. That’s a bit much for me, is it possible to use fewer cells?
Solution: To use fewer cells per run you have two options. You can load cells at a higher dilution than recommended (300 cells / µl) or you can load a smaller volume of cells (min. 100 µl, max. 250 µl). To take into account the smaller volume of cell suspension, the user should reduce the total length of the run by changing the time parameters on the Nadia Innovate.
Problem: I would like to encapsulate my cells in hydrogel, which is temperature sensitive. How can I do that?
Solution: To ensure that hydrogel does not solidify during encapsulation, you can set the Nadia Innovate at your chosen temperature, between 4 and 40°C.
This list is only a short insight into the solutions offered with the Nadia Innovate. Do you have a specific single cell research question or problem you would like to solve? Get in touch and speak to one of our single cell specialists today!
For more information….
Consult our ‘Get Started’ guide. The guide walks researchers through every stage of protocol optimization. It includes a workflow that leads the user in a stepwise manner through protocol development and single cell customization.
Developing or optimizing single cell protocols can be daunting, especially when one is not familiar with the basic principles of microfluidics. But don’t worry, at Dolomite Bio we are here to help.
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