A Single Cell Extraction Chip Using Vibration-Induced Whirling Flow and a Thermo-Responsive Gel Pattern

We propose a single cell extraction chip with an open structure, which utilizes vibration-induced whirling flow and a single cell catcher. By applying a circular vibration to a micropillar array spiral pattern, a whirling flow is induced around the micropillars, and target cells are transported towards the single cell catcher placed at the center of the spiral. The single cell catcher is composed of a single-cell-sized hole pattern of thermo-responsive gel. The gel swells at low temperatures (≲32 ◦C) and shrinks at high temperatures (≳32 ◦C), therefore, its volume expansion can be controlled by an integrated microheater. When the microheater is turned on, a single cell is trapped by the hole pattern of the single cell catcher. Then, when the microheater is turned off, the single cell catcher is cooled by the ambient temperature. The gel swells at this temperature, and the hole closes to catch the single cell. The caught cell can then be released into culture wells on a microtiter plate by heating the gel again. We conducted single cell extraction with the proposed chip and achieved a 60% success rate, of which 61% cells yielded live cells.

[1]  Fumihito Arai,et al.  Local streamline generation by mechanical oscillation in a microfluidic chip for noncontact cell manipulations , 2012 .

[2]  Leslie Y Yeo,et al.  Ultrafast microfluidics using surface acoustic waves. , 2009, Biomicrofluidics.

[3]  A. Ashkin,et al.  Optical trapping and manipulation of viruses and bacteria. , 1987, Science.

[4]  Jarrett Rosenberg,et al.  Single cell transcriptional profiling reveals heterogeneity of human induced pluripotent stem cells. , 2011, The Journal of clinical investigation.

[5]  Peng Li,et al.  Surface acoustic wave microfluidics. , 2013, Lab on a chip.

[6]  Dino Di Carlo,et al.  Automated cellular sample preparation using a Centrifuge-on-a-Chip. , 2011, Lab on a chip.

[7]  T. Fukuda,et al.  On chip single-cell separation and immobilization using optical tweezers and thermosensitive hydrogel. , 2005, Lab on a chip.

[8]  R G Sweet,et al.  Fluorescence Activated Cell Sorting , 2020, Definitions.

[9]  Fumihito Arai,et al.  Isolation and extraction of target microbes using thermal sol-gel transformation. , 2003, The Analyst.

[10]  Ming C. Wu,et al.  Massively parallel manipulation of single cells and microparticles using optical images , 2005, Nature.

[11]  B. Drachman,et al.  The steady streaming generated by a vibrating plate parallel to a fixed plate , 1982 .

[12]  Mattias Goksör,et al.  A microfluidic device for reversible environmental changes around single cells using optical tweezers for cell selection and positioning. , 2010, Lab on a chip.

[13]  S. Takayama,et al.  Microfluidics for flow cytometric analysis of cells and particles , 2005, Physiological measurement.

[14]  S. Bodovitz,et al.  Single cell analysis: the new frontier in 'omics'. , 2010, Trends in biotechnology.

[15]  Barry R Lutz,et al.  Hydrodynamic tweezers: 1. Noncontact trapping of single cells using steady streaming microeddies. , 2006, Analytical chemistry.

[16]  Fumihito Arai,et al.  On-Chip Gel-Valve Using Photoprocessable Thermoresponsive Gel , 2014 .

[17]  Yoshiyuki Yokoyama,et al.  Micro- and Nano-fabrication of Stimulus-responsive Polymer using Nanoimprint Lithography , 2011 .

[18]  I. Johnsen,et al.  Boundary Layer Flow near a Cylindrical Obstacle in an Oscillating Incompressible Fluid , 1954 .

[19]  R. Pethig.  Review article-dielectrophoresis: status of the theory, technology, and applications. , 2010, Biomicrofluidics.

[20]  Fumihito Arai,et al.  High‐speed separation system of randomly suspended single living cells by laser trap and dielectrophoresis , 2001, Electrophoresis.

[21]  Shoji Takeuchi,et al.  A trap-and-release integrated microfluidic system for dynamic microarray applications , 2007, Proceedings of the National Academy of Sciences.

[22]  M. Yamada,et al.  Pinched flow fractionation: continuous size separation of particles utilizing a laminar flow profile in a pinched microchannel. , 2004, Analytical chemistry.

[23]  Mehmet Toner,et al.  A microfabrication-based dynamic array cytometer. , 2002, Analytical chemistry.

[24]  S. Quake,et al.  A microfabricated fluorescence-activated cell sorter , 1999, Nature Biotechnology.

[25]  W. Bonner,et al.  Cell Sorting: Automated Separation of Mammalian Cells as a Function of Intracellular Fluorescence , 1969, Science.