Molecular machines have previously been designed that are propelled by DNAzymes1–3, protein enzymes4–6 and strand-displacement7–9. These engineered machines typically move along precisely defined one- and two-dimensional tracks. Here, we report a DNA walker that uses hybridisation to drive walking on DNA-coated microparticle surfaces. Through purely DNA:DNA hybridisation reactions, the nanoscale movements of the walker can lead to the generation of a single-stranded product and the subsequent immobilisation of fluorescent labels on the microparticle surface. This suggests that the system could be of use in analytical and diagnostic applications, similar to how strand exchange reactions in solution have been used for transducing and quantifying signals from isothermal molecular amplification assays10,11. The walking behaviour is robust and the walker can take more than 30 continuous steps. The traversal of an unprogrammed, inhomogeneous surface is also due entirely to autonomous decisions made by the walker, behaviour analogous to amorphous chemical reaction network computations12,13 that have been shown to lead to pattern formation14–17.
Background Detection of a small number of circulating tumor cells is important, especially at the early stages of cancer. The small number of CTCs is hard to detect as very few approaches are sensitive enough to differentiate these from the pool of other cells. Improving the affinity of a selective surface-functionalized molecule is important given the sparsity of CTCs in vivo. There are a number of proteins and aptamers that provide such a high affinity but using a surface nano-texturing increases this affinity even further. Method This work reports an approach to improve affinity of tumor cell capture by using novel aptamers against cell-membrane over-expressed Epidermal Growth Factor Receptors (EGFR) on a nano-textured polydimethylsiloxane (PDMS) substrate. Surface immobilized aptamers are used to specifically capture tumor cells from physiological samples. Results The nano-texturing of PDMS increased surface roughness at the nanoscale. This increased the effective surface area and resulted in a significantly higher degree of surface functionalization. The phenomenon resulted in increased density of immobilized EGFR specific RNA aptamer molecules and provided significantly higher efficiency to capture cancer cells from a mixture. The data showed that CTCs could be captured and enriched leading to higher yield, yet higher background. Conclusion The comparison of glass slides, plain PDMS and nano-textured PDMS functionalized with aptamers show that a two-fold approach of using aptamers on nano-textured PDMS can be an important factor for cancer cytology devices especially for the idea of lab-on-chip towards higher yield in capture efficiency.
Readily programmable chemical networks are important tools as the scope of chemistry expands from individual molecules to larger molecular systems. While many complex systems have been constructed using conventional organic and inorganic chemistry, the programmability of biological molecules such as nucleic acids allows for precise, high-throughput, and automated design, as well as simple, rapid, and robust implementation. Here we show that systematic and quantitative control over the diffusivity and reactivity of DNA molecules yields highly programmable chemical reaction networks (CRNs) that execute at the macroscale. In particular, we design and implement non-enzymatic DNA circuits capable of performing pattern transformation algorithms such as edge detection. We also show that it is possible to fine-tune and multiplex such circuits. We believe these strategies will provide programmable platforms for prototyping CRNs, for discovering bottom-up construction principles, and for generating patterns in materials.
This paper describes a method for manipulating the temperature inside aqueous droplets, utilizing a thermoelectric cooler to control the temperature of select portions of a microfluidic chip. To illustrate the adaptability of this approach, we have generated an "ice valve" to stop fluid flow in a microchannel. By taking advantage of the vastly different freezing points for aqueous solutions and immiscible oils, we froze a stream of aqueous droplets that were formed on-chip. By integrating this technique with cell encapsulation into aqueous droplets, we were also able to freeze single cells encased in flowing droplets. Using a live-dead stain, we confirmed the viability of cells was not adversely affected by the process of freezing in aqueous droplets provided cryoprotectants were utilized. When combined with current droplet methodologies, this technology has the potential to both selectively heat and cool portions of a chip for a variety of droplet-related applications, such as freezing, temperature cycling, sample archiving, and controlling reaction kinetics.
Early detection and isolation of circulating tumor cells (CTC) can enable better prognosis for cancer patients. A Hele-Shaw device with aptamer functionalized glass beads is designed, modeled, and fabricated to efficiently isolate cancer cells from a cellular mixture. The glass beads are functionalized with anti-epidermal growth factor receptor (EGFR) aptamer and sit in ordered array of pits in PDMS channel. A PDMS encapsulation is then used to cover the channel and flow through cell solution. The beads capture cancer cells from flowing solution depicting high selectivity. The cell-bound glass beads are then re-suspended from the device surface followed by the release of 92% cells from glass beads using combination of soft shaking and anti-sense RNA. This approach ensures that the cells remain in native state and undisturbed during capture, isolation and elution for post-analysis. The use of highly selective anti-EGFR aptamer with the glass beads in an array and subsequent release of cells with antisense molecules provide multiple levels of binding and release opportunities that can help in defining new classes of CTC enumeration devices.
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