Tie Yang

A Comprehensive Review of Optical Stretcher for Cell Mechanical Characterization at Single-Cell Level

This paper presents a comprehensive review of the development of the optical stretcher, a powerful optofluidic device for single cell mechanical study by using optical force induced cell stretching. The different techniques and the different materials for the fabrication of the optical stretcher are first summarized. A short description of the optical-stretching mechanism is then given, highlighting the optical force calculation and the cell optical deformability characterization. Subsequently, the implementations of the optical stretcher in various cell-mechanics studies are shown on different types of cells. Afterwards, two new advancements on optical stretcher applications are also introduced: the active cell sorting based on cell mechanical characterization and the temperature effect on cell stretching measurement from laser-induced heating. Two examples of new functionalities developed with the optical stretcher are also included. Finally, the current major limitation and the future development possibilities are discussed.

A comprehensive strategy for the analysis of acoustic compressibility and optical deformability on single cells

We realized an integrated microfluidic chip that allows measuring both optical deformability and acoustic compressibility on single cells, by optical stretching and acoustophoresis experiments respectively. Additionally, we propose a measurement protocol that allows evaluating the experimental apparatus parameters before performing the cell-characterization experiments, including a non-destructive method to characterize the optical force distribution inside the microchannel. The chip was used to study important cell-mechanics parameters in two human breast cancer cell lines, MCF7 and MDA-MB231. Results indicate that MDA-MB231 has both higher acoustic compressibility and higher optical deformability than MCF7, but statistical analysis shows that optical deformability and acoustic compressibility are not correlated parameters. This result suggests the possibility to use them to analyze the response of different cellular structures. We also demonstrate that it is possible to perform both measurements on a single cell, and that the order of the two experiments does not affect the retrieved values.

Integrated Optofluidic Chip for Low-Volume Fluid Viscosity Measurement

In the present work, an integrated optofluidic chip for fluid viscosity measurements in the range from 1 mPa·s to 100 mPa·s is proposed. The device allows the use of small sample volumes (<1 µL) and the measurement of viscosity as a function of temperature. Thanks to the precise control of the force exerted on dielectric spheres by optical beams, the viscosity of fluids is assessed by comparing the experimentally observed movement of dielectric beads produced by the optical forces with that expected by numerical calculations. The chip and the developed technique are validated by analyzing several fluids, such as Milli-Q water, ethanol and water–glycerol mixtures. The results show a good agreement between the experimental values and those reported in the literature. The extremely reduced volume of the sample required and the high flexibility of this technique make it a good candidate for measuring a wide range of viscosity values as well as for the analysis of nonlinear viscosity in complex fluids.

Sorting on the basis of deformability of single cells in a femtosecond laser fabricated optofluidic device

Optical stretching is a powerful technique for the mechanical phenotyping of single suspended cells that exploits cell deformability as an inherent functional marker. Dual-beam optical trapping and stretching of cells is a recognized tool to investigate their viscoelastic properties. The optical stretcher has the ability to deform cells through optical forces without physical contact or bead attachment. In addition, it is the only method that can be combined with microfluidic delivery, allowing for the serial, high-throughput measurement of the optical deformability and the selective sorting of single specific cells. Femtosecond laser micromachining can fabricate in the same chip both the microfluidic channel and the optical waveguides, producing a monolithic device with a very precise alignment between the components and very low sensitivity to external perturbations. Femtosecond laser irradiation in a fused silica chip followed by chemical etching in hydrofluoric acid has been used to fabricate the microfluidic channels where the cells move by pressure-driven flow. With the same femtosecond laser source two optical waveguides, orthogonal to the microfluidic channel and opposing each other, have been written inside the chip. Here we present an optimized writing process that provides improved wall roughness of the micro-channels allowing high-quality imaging. In addition, we will show results on cell sorting on the basis of mechanical properties in the same device: the different deformability exhibited by metastatic and tumorigenic cells has been exploited to obtain a metastasis-cells enriched sample. The enrichment is verified by exploiting, after cells collection, fluorescence microscopy.

A micro-opto-acousto-fluidic chip for single cell mechanics evaluation

The rapid development of microfluidic circuits and lab-on-chip devices for cell studies opened new perspectives for cellular biology, in particular for the biophysics and biomechanics of single cells. At the state of the art, a lot of attention is being paid to the possible integration of optical and acoustic actuators within microfluidic system because they both provide a contactless way for cell manipulation and analysis; additionally, they both offer other advantages such as rapid actuation, programmable capability and simple operation.

Rheological study of a DNA transient network by optophoresis

Micro-rheology recently emerged as a promising technique for investigating the viscoelastic properties of complex fluids at a molecular scale. The study of the rheological properties of polymer solutions and reversible gels is more complex than that of molecular fluids, because these fluids commonly exhibit nonlinear viscosity behaviour. The research activity that we report about has been dedicated to the analysis of the dynamic behaviour of a DNA-particles system. DNA particles shaped as nano-stars with sticky ending points are suspended in water, and by lowering the temperature, below a gelation threshold, the nano-stars stick one to each other to form an hydrogel [1, 2].

Erratum: Corrigendum: A comprehensive strategy for the analysis of acoustic compressibility and optical deformability on single cells

Scientific Reports 6: Article number: 23946; published online: 04 April 2016; updated: 19 January 2017. This Article contains errors. In Figure 3c, the y-axis ‘Acoustic compressibility (10−10 Pa−1)’ is incorrectly given as ‘Acoustic compressibility (TPa−1)’. The correct Figure 3c appears below as Figure 1.

Femtosecond laser fabricated microfluorescence-activated cell sorter for single cell recovery

Manipulation, sorting and recovering of specific live cells from samples containing less than a few thousand cells is becoming a major hurdle in rare cell exploration such as stem cell research or cell based diagnostics. Moreover the possibility of recovering single specific cells for culturing and further analysis would be of great impact in many biological fields ranging from regenerative medicine to cancer therapy. In recent years considerable effort has been devoted to the development of integrated and low-cost optofluidic devices able to handle single cells, which usually rely on microfluidic circuits that guarantee a controlled flow of the cells. Among the different microfabrication technologies, femtosecond laser micromachining (FLM) is ideally suited for this purpose as it provides the integration of both microfluidic and optical functions on the same glass chip leading to monolithic, robust and portable devices. Here a new optofluidic device is presented, which is capable of sorting and recovering of single cells, through optical forces, on the basis of their fluorescence and. Both fluorescence detection and single cell sorting functions are integrated in the microfluidic chip by FLM. The device, which is specifically designed to operate with a limited amount of cells but with a very high selectivity, is fabricated by a two-step process that includes femtosecond laser irradiation followed by chemical etching. The capability of the device to act as a micro fluorescence-activated cell sorter has been tested on polystyrene beads and on tumor cells and the results on the single live cell recovery are reported.