A novel method for the isolation of single cells mimicking circulating tumour cells adhered on Smart Bio Surface slides by Laser Capture Microdissection.

In recent years, the importance of isolating single cells from blood circulation for several applications, such as non-invasive tumour diagnosis, the monitoring of minimal residual disease, and the analysis of circulating fetal cells for prenatal diagnosis, urged the need to set up innovative methods. For such applications, different methods were developed. All show some weaknesses, especially a limited sensitivity, and specificity. Here we present a new method for isolating a single or a limited number of cells adhered to SBS slides (Tethis S.p.a.) (a glass slide coated with Nanostructured Titanium Dioxide) by Laser Capture Microdissection (LCM) and subsequent Whole Genome Amplification. SBS slides have been shown to have an optimal performance in immobilizing circulating tumour cells (CTCs) from early breast cancer patients. In this work, we spiked cancer cells in blood samples to mimic CTCs. By defining laser parameters to cut intact samples, we were able to isolate genetically intact single cells. We demonstrate that SBS slides are optimally suited for isolating cells using LCM and that this method provides high-quality DNA, ideal for gene-specific assays such as PCR and Sanger sequencing for mutation analysis.

Disposable Optical Stretcher Fabricated by Microinjection Moulding.

Microinjection moulding combined with the use of removable inserts is one of the most promising manufacturing processes for microfluidic devices, such as lab-on-chip, that have the potential to revolutionize the healthcare and diagnosis systems. In this work, we have designed, fabricated and tested a compact and disposable plastic optical stretcher. To produce the mould inserts, two micro manufacturing technologies have been used. Micro electro discharge machining (µEDM) was used to reproduce the inverse of the capillary tube connection characterized by elevated aspect ratio. The high accuracy of femtosecond laser micromachining (FLM) was exploited to manufacture the insert with perfectly aligned microfluidic channels and fibre slots, facilitating the final composition of the optical manipulation device. The optical stretcher operation was tested using microbeads and red blood cells solutions. The prototype presented in this work demonstrates the feasibility of this approach, which should guarantee real mass production of ready-to-use lab-on-chip devices.

Rapid Prototyping of Plastic Lab-on-a-Chip by Femtosecond Laser Micromachining and Removable Insert Microinjection Molding.

We have introduced a new hybrid fabrication method for lab-on-a-chip devices through the combination of femtosecond laser micromachining and removable insert micro-injection molding. This method is particularly suited for the fast prototyping of new devices, while maintaining a competitive low cost. To demonstrate the effectiveness of our approach, we designed, fabricated, and tested a completely integrated flow cytometer coupled to a portable media device. The system operation was tested with fluorescent plastic micro-bead solutions ranging from 100 beads/µL to 500 beads/µL. We demonstrated that this hybrid lab-on-a-chip fabrication technology is suitable for producing low-cost and portable biological microsystems and for effectively bridging the gap between new device concepts and their mass production.