Semi-permeable capsules

Accelerate Your Multi‑Step Molecular Analysis With Semi‑Permeable Capsules

Semi-permeable capsules are a breakthrough technology designed to support multi-step molecular analysis, even those that require a harsh lysis step, without loss of single-cell compartmentalization. With the push of a single button, up to 8 samples can be simultaneously incorporated into biocompatible SPCs, encapsulating tens of thousands of single cells into SPCs in a matter of minutes. Once generated, the SPCs are stable under any reaction conditions and are suitable for any downstream reaction, which can be performed in a bulk-like manner on millions of SPCs in parallel using standard laboratory equipment.

Expand Your Capabilities With Multi‑Step Workflows

Semi-Permeable Capsule compartments have an aqueous core that is surrounded by a hydrogel shell. The shell functions like a size-selective membrane and enables an unrestricted exchanges of small reaction components (e.g. enzymes, detergents, salts) while retaining large molecules, such as mRNA and genomic DNA, and cells inside the compartments. Once generated, the SPC compartments are stable under any reaction conditions thus enabling multi-step workflows.

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Applications

Use even the harshest lysis conditions to access bacterial genomes without disrupting the capsule integrity or affecting downstream reactions. Semi-permeable capsules overcome the primary drawbacks of traditional water-in-oil droplets by enabling the unrestricted exchange of small reaction components, such as enzymes, detergents, and salts, while maintaining compartmentalization of single cells. Changing reaction composition between each step is as easy as spinning the SPCs down with a table-top centrifuge and changing the reaction buffer with a pipettor.

Perform high-throughput transcriptomic profiling on thousands of single cells using an inherently scalable cell barcoding approach in semi-permeable capsules. Unlike traditional water-in-oil droplets, SPCs are stable under any reaction conditions and allow easy reagent exchange, which means that washes can be used between steps to create the optimal conditions for downstream reactions such as ligation, reverse transcription, and PCR. Sequencing-ready libraries are released under mild conditions, which may improve RNA quality and reduce the variability in transcriptome capture.

Enhance PCR efficiency and increase target coverage for better high-throughput single-cell targeted sequencing. SPCs allow for the efficient transfer of reagents and small molecules, such as primers and dNTPs, during reaction steps and act as a size-selective membrane to prevent the loss of DNA fragments during PCR amplification. Massive parallelization with SPCs permits multiplexed assays, where different primers or probes can be used in different capsules to simultaneously amplify different target regions, allowing for more efficient use of sequencing resources.

Significantly reduce the cost and time required for single-cell long-read sequencing with semi-permeable capsules, which are inherently scalable and can be processed in parallel or multiplexed for better utilization of sequencing capacity. SPCs are stable in any reaction conditions and, compared to traditional water-in-oil droplets, are less likely to break or merge during long-read sequencing. The capsules act as a size-selective membrane and permit small molecule exchange, which enables more precise control over the reaction contents while simultaneously preventing the loss of large DNA fragments, leading to more accurate and complete sequencing results.

Profile the gene expression of thousands of individual cells simultaneously using a fluorescent readout, as SPCs are compatible with traditional fluorescence-assisted cell sorting (FACS) devices and epifluorescence microscopy. The size-selective membrane retains RNA molecules while allowing for the removal of RNases and other small molecules, resulting in efficient RNA purification. By incorporating a multiplex selection of fluorescently-labeled primers in the RT-PCR reaction mix, the expression of the selected genes of interest can be profiled and quantified in a high-throughput manner.

Increase the throughput of colony-based assays by cultivating thousands of microorganisms in parallel while maintaining colony isolation. SPCs are biocompatible and feature a size-selective hydrogel shell that retains cells within the compartments while allowing easy liquid media exchange. Single-cell encapsulation provides a protective environment for colonies to grow without competition from others, which is especially beneficial for slow-growing species.