Future Applications
Sphere Fluidics’ picodroplet technology has many advantages, once of which is its versatility. While we know our research platforms such as Cyto-Mine® can accelerate antibody discovery and cell line development workflows, there are also other potential areas where our novel technologies can potentially be applied in the future. Please see some of our existing work and potential future application areas below. Is your area of interest not listed? Get in touch.
Cell Therapy Engineering and Precision Genome Editing
Stem cells and T-cells can be engineered to differentiate and form specific cell types or phenotypes respectively. However, the identification of the factors that cause reproducible differentiation and the analysis of the newly generated cells themselves can be challenging.
At Sphere Fluidics, we’re dedicated to making these challenges simple. Our unique platforms enable you to screen and isolate, and in the future potentially precision engineer, individual cells using automated, miniaturised and integrated techniques that are significantly faster and more cost-effective than alternatives.
Our Unique Benefits for Cell Therapy Engineering and Genome Editing
Picodroplets provide a unique way of compartmentalising single cells for individual engineering, screening and detailed analysis. Once compartmentalised, these picodroplets can be fused with different factors (e.g. genome editing complexes, single viruses, growth factors, cDNAs, 3D substrata, or engineered micro-gels) to stimulate or transform the cells, which can then be sorted and assayed to isolate specific phenotypes.
Since you can compartmentalise cells using our picodroplet technology, each one can then be engineered on a single cell basis. This removes the biases and inefficiencies seen with the transfection of bulk populations of cells. Our technology also has significant potential for use in the clinic, for example, in the precision reprogramming of T-cells collected specifically from a patient to create personalised medicines. Potentially, we can also minimise the risk of cell damage or oncogenicity by controlling the exact number of genome editing complexes, cDNAs or viruses we introduce into each cell – even to the single molecule level.
Our Technology in Action for Cell Therapy Engineering and Genome Editing
Our picodroplet technology can be used for numerous single cell research applications, including:
- Controlling the transfection and selection of transformed T-cells for cell therapy
- Automation and miniaturisation of genome editing
- Separation of specific phenotypes
- Investigating the impact of cell-cell or cell-molecule interactions on cellular phenotype
- Identifying cell instability and variation
- Studying the differentiation process of single cells
As can be seen in the diagram, our systems provide a simple, fast and high-throughput process. For example, with T-cells, once blood is collected from a patient, target cells are isolated and purified. These cells are then encapsulated into individual picodroplets, which are fused with another picodroplet containing the factor of interest (e.g. a viral construct) and electroporated.
Following incubation and sorting, the positive clones or ‘hits’ can then be analysed by DNA/RNA sequencing, proteomic analysis or other phenotyping tools.
In the application of transfection and selection of transformed T-cells for cell therapy, the automation and high precision of our systems will improve cell separation and the overall transduction process. Since this approach compartmentalises cells, they can be collected from a patient, separated into single picodroplets and reprogrammed. This should have a high efficiency meaning that less sample is needed from the patient and generation of a more effective precision therapy.
To find out how you could utilise our technology for cell engineering and genome editing in your lab, get in touch.
Antibiotic Resistance Analysis and Antibiotic Discovery
Over the last twenty years, anti-microbial resistance has become a serious threat to public health. It can occur at a rate of approximately one in one billion bacteria. Recently, there’s been a resurgent demand for the development of new antibiotics. Current techniques for antibiotic discovery are limited by antibiotic diversity, compound supply and potency, as well as the time, cost and labour investment required in such studies.
We’ve developed our picodroplet technology with speed and efficiency in mind. By miniaturizing the screening process, you can increase throughput while still obtaining accurate and insightful data. We’re confident that our instruments can help make antibiotic discovery and resistance analysis exciting and fruitful areas of research once again.
The Unique Benefits of Our Systems for Research Into Antibiotic Resistance and Antibiotic Discovery
With the demand for faster and more effective routes to antibiotic discovery, we’ve built our systems to make it feasible to screen more than one billion bacteria for signs of antibiotic resistance.
After screening in the presence of antibiotics, the surviving bacteria can be retrieved using optical readouts and then sequenced. This analysis allows you to identify targets of interest that can be used to explore the genetic basis for antibiotic resistance. We can also compartmentalise single microbes (e.g. Streptomycetes or Actinomycetes) and ensure that both slow-growing and fast-growing bacteria can replicate without competition and thus produce diverse antibiotics that are usually not seen in mixed bulk populations. These antibiotic-producing microbes can be co-compartmentalised with target microbes and the level of target cell killing or damage is detected optically using fluorescent probes or by modulation of expression. e.g. GFP.
As demonstrated in the graph below, a single E. coli microbe can grow through to circa 7,000 in 8 hours. This equates to around one billion bacteria per biochip – more than adequate to detect the emergence of antibiotic-resistance. This high throughput model saves you time, as well as associated reagent costs and reduces the generation of environmental waste usually generated with microtitre plate based techniques.
Learn More About Our Technology and its Benefits
Our Systems in Action for Antibiotic Resistance and Antibiotic Discovery
Our technology can be applied to antibiotic resistance research through the analysis of bacterial cells. In an experiment, more than one billion bacteria were tested for antibiotic resistance using our picodroplet technology. Individual bacteria were encapsulated in picodroplets along with an antibiotic and incubated at 37°C. While test antibiotic concentrations prevented most bacteria from growing, the few that did manage to proliferate were clearly antibiotic-resistant. These samples were then taken on for genetic sequencing and morphological studies.
As can be seen in the figure, a total of 124 antibiotic-resistant colonies were observed on 10 antibiotic-containing plates, representing a mutation rate of c. 1 in ten million. Following this, the survivors were sub-cultured for further downstream analysis by sequencing and other techniques.
