Gabriel Dumy– gabriel.dumy@espci.fr
Mauricio Hoyos – mauricio.hoyos@espci.fr
Jean-Luc Aider – jean-luc.aider@espci.fr
ESPCI Paris – PMMH Lab
10 rue Vauquelin
Paris, 75005, FRANCE

Popular version of paper 3aPA7, “Investigation on a novel photoacoustofluidic effect”

Presented Wednesday morning, December 6, 2017, 11:00-11:15 AM, Balcony L

174th ASA Meeting, New Orleans

Amongst the various ways of manipulating suspensions, acoustic levitation is one of the most practical yet not very known to the public. Allowing for contactless concentration of microscopic bodies (from particles to living cells) in fluids (whether it be air, water, blood…), this technique only requires a small amount of power and materials. It is thus smaller and less power consuming than other technologies using magnetic or electric fields for instance and does not require any preliminary tagging.

Acoustic levitation occurs when using standing ultrasonic waves trapped between two reflecting walls. If the ultrasonic wavelength _ac is matched to the distance between the two walls (it has to be a certain number of the half wavelength), then an acoustic pressure field forces the particles or cells to move toward the region where the acoustic pressure is minimal (this region is called a pressure node) [1]. Once the particles or cells have reached the pressure node, they can be kept in so-called “acoustic levitation” as long as needed. They are literally trapped in an “acoustic tweezer”. Using this method, it is easy to force cells or particles to create large clusters or aggregates than can be kept in acoustic levitation as long as the ultrasonic field is on.

What happens if we illuminate the aforementioned aggregates of fluorescent particles or cells with a strong monochromatic (only one color) optic wave? If this wave is absorbed by the levitating objects, then the previously very stable aggregate explodes.

We can observe that the particles are now ejected from the illuminated aggregate at great speed from its periphery. But they are still kept in acoustic levitation, which is not affected by the introduction of light.

We determined that the key parameter is the absorption of light by the levitating objects because the explosions happened even with non-fluorescent particles. Moreover, this phenomenon exhibits a strong coupling between light and sound, as it needs the two sources of energy to be present at the same time to occur. If the particles are not in acoustic levitation, on the bottom of the cavity or floating in the suspending medium, even a very strong light does not move them. Without the adequate illumination, we only observe a classical acoustic aggregation process.

Using this light absorption property together with acoustic levitation opens the way to more complex and challenging experiments, like advanced manipulations of micro-objects in acoustic levitation or fast and highly selective sorting of mixed suspensions, since we can discriminate these particles not only on their mechanical properties but also on their optic ones.

We did preliminary experiments with living cells. We observed that human red blood cells (RBCs), having a strong absorption of blue light, could be easily manipulated by both sounds and light. We were able to break up RBCs aggregates very quickly. As a matter of fact, this new effect coupling both acoustics and light suggests all new perspectives for living cells manipulation and sorting, like cell washing (removing unwanted cells from the target cell).  Indeed, most of the living cells absorb light at different wavelengths and can already be manipulated using acoustic fields. This discovery should allow very selective manipulations and/or sorting of living cells in a very simple and easy way, using a low-cost setup.

Figure 1. Illustration of the acoustic manipulation of suspensions. A suspension is first focused under the influence of the vertical acoustic pressure field in red (a and b). Once in the pressure node, the suspension is radially aggregated c) by secondary acoustic forces [2]. On d), when we enlighten the stable aggregate with an adequate wavelength, this one laterally explodes.

Figure 2. (videos missing): Explosion (red_explosion) of the previously formed aggregate of 1.6 polystyrene beads, that are red fluorescent, by a green light. Explosion (green_explosion) of an aggregate of 1.7µm green fluorescent polystyrene beads by a blue light.

Figure 3 (videos missing): Illustration of the separation potential of the phenomenon. We take an aggregate (a) that is a mix of two kind of polystyrene particles with same diameter, one absorbing blue light and fluorescing green (b), the other absorbing green light and fluorescing red (c), that we cannot separate by acoustics alone. We expose this aggregate to blue light for 10 seconds. On the bottom row is shown the effect of this light, we effectively separated the blue absorbing particles (e) from the green absorbing one (f).

Movie missing – describes the observation from the top of the regular acoustic aggregation process of a suspension of 1.6µm polystyrene beads.

[1] K. Yosioka and Y. Kawasima, “Acoustic radiation pressure on a compressible sphere,” Acustica, vol. 5, pp. 167–173, 1955.

[2] G. Whitworth, M. A. Grundy, and W. T. Coakley, “Transport and harvesting of suspended particles using modulated ultrasound,” Ultrasonics, vol. 29, pp. 439–444, 1991.

Xiaoyun Ding- Xiaoyun.Ding@Colorado.edu
Department of Mechanical engineering
University of Colorado at Boulder
Boulder, CO 80309

Popular version of paper 3aPA3, “Standing Surface Acoustic Wave Enabled Acoustofluidics for Bioparticle Manipulation”
Presented Wednesday, December 06, 2017, 9:30-10:00 AM, Balcony L
174th ASA meeting, New Orleans

Techniques that can noninvasively and dexterously manipulate cells and other bioparticles (such as organisms, DNAs, proteins, and viruses) in a compact system are invaluable for many applications in life sciences and medicine. Historically, optical tweezers have been the primary tool used in the scientific community for bioparticle manipulation. Despite the remarkable capability and success, optical tweezers have notable limitations, such as complex and bulky instrumentation, high equipment costs, and low throughput. To overcome the limitations of optical tweezers and other particle manipulation methods, we have developed a series of acoustic-based, on-chip devices (Figure to the left) called acoustic tweezers that can manipulate cells and other bioparticles using sound waves in microfluidic channel. Cells viability and proliferation assays were also conducted to confirm the non-invasiveness of our technique. The simple structure/setup of these acoustic tweezers can be integrated with a small radio-frequency power supply and basic electronics to function as a fully integrated, portable, and inexpensive cell-manipulation system. Along with my colleagues, I have demonstrated that our acoustic tweezers can achieve the following functions: 1) single cell/organism manipulation [1]; 2) high-efficiency cell separation [2]; and 3) multichannel cell sorting [3].

Acoustic tweezers based single cell/organism manipulation
The acoustic tweezers I developed was the first acoustic manipulation method which can trap and dexterously manipulate single microparticles, cells, and entire organisms (i.e., Caenorhabditis elegans) along a programmed route in two-dimensions within a microfluidic chip [1]. We demonstrate that the acoustic tweezers can move a 10-µm single polystyrene bead to write the word “PNAS” and a bovine red blood cell to trace the letters “PSU” (Figure to the right). It was also the first technology capable of touchless trapping and manipulating Caenorhabditis elegans, a one-millimeter long roundworm that is one of the most important model systems for studying diseases and development in humans. To the best of our knowledge, this is the first demonstration of non-invasive, non-contact manipulation of C. elegans, a function that is challenging for optical tweezers.

Acoustic tweezers based high-efficiency cell separation
Simple and high-efficiency cell separation techniques are fundamentally important in biological and chemical analyses such as cancer cell detection, drug screening, and tissue engineering. In particular, the ability to separate cancer cells (such as leukaemia cells) from human blood can be invaluable for cancer biology, diagnostics, and therapeutics. We have developed an standing surface acoustic wave based cell separation technique that can achieve high-efficiency (>95%) separation of human leukemia cells (HL-60) from human blood cells and high efficiency separation of breast  cancer cells from human blood based on their size difference (Figure to the right). This method is simple and versatile, capable of separating virtually all kinds of cells (regardless of charge/polarization or optical properties) with high separation efficiency and low power consummation.

Acoustic tweezers based multichannel cell sorting
Cell sorting is essential for many fundamental cell studies, cancer research, clinical medicine, and transplantation immunology. I developed an acoustic-based method that can precisely sort cell into five separate outlets of cells (Figure to the right), rendering it particularly desirable for multi-type cell sorting [3]. Our device requires small sample volumes (~100 μl), making it an ideal tool for research labs and point-of-care diagnostics. Furthermore, it can be conveniently integrated with a small power supply, a fluorescent detection module, and a high-speed electrical feedback module to function as a fully integrated, portable, inexpensive, multi-color, miniature fluorescence-activated cell sorting (μFACS) system.

 References:

1. Xiaoyun Ding, et al., On-Chip Manipulation of Single Microparticles, Cells, and Organisms Using Surface Acoustic Waves, Proceedings of the National Academy of Sciences of the United States of America (PNAS), 2012, 109, 11105-09,.
2. Xiaoyun Ding, et al., Cell separation using tilted-angle standing surface acoustic waves, Proceedings of the National Academy of Sciences of the United States of America (PNAS), 111, 12992-12997 (2014).
3. Xiaoyun Ding, et al., Standing surface acoustic wave (SSAW) based multichannel cell sorting, Lab On a Chip, 2012,12, 4228–31,. (COVER ARTICLE)
4. Xiaoyun Ding, et al., Lab on a Chip, 2012, 12, 2491-97. (COVER ARTICLE)