We focus on the development and fabrication of SU8TM-based microchannel networks, which can be integrated into microdevices for fast drug delivery and cell transport on chips. Instead of using sacrificial materials or wafer bonding, a new simplified fabrication method is developed. Single- and double-layered SU8TM channels on silicon substrates are successfully achieved by using this new method, as well as integration of these SU8 channels with microelectrode arrays. A series of cell transport experiments is also successfully performed on these devices. This new fabrication approach and the resulting cell transport experiments are discussed in detail.
This paper focuses on the development of two MEMS-based devices for lab-on-a-chip bio-applications. The first device is designed to facilitate cell secretion studies by enabling parallel electrochemical detection with millisecond resolution. Initial prototypes of micro-arrays have been fabricated with Cr/Au microelectrodes on various substrates such as polyimide, SU-8 and SiO2. An FT cell-line (bullfrog fibroblast, American Tissue Culture Collection) has been successfully established and cultured directly on these prototype micro-arrays. It is well known that the FT cells can uptake hormones or other macromolecules from the culture media through a non-specific uptake mechanism which is still under investigation. After culturing on micro-arrays, FT cells were loaded with norepinephrine of various concentrations by incubation in the culture media supplied with norepinephrines. Rapid elevation of intracellular Ca2+ levels triggers the exocytosis of norepinephrine which then can be detected by the Cr/Au electrodes. Microfabrication of these prototype micro-arrays as well as cell culture and electrochemical detection results will be presented in this paper. The second device is designed for 3-dimensional transportation of living cells on chips. Initial prototypes of micro-arrays were fabricated with SU-8 buried channels on a silicon substrate. Both single-layered and double-layered SU-8 buried channels have been realized to enable 2D and 3D cell transportation. Stained solutions were used to visualize fluid transport through the channel networks. Following this, living FT cells in solution were successfully transported through single-layered SU-8 channels. Testing of 3D transportation of living FT cells is underway. Microfabrication of these prototype micro-arrays and living cell transportation on chips will also be presented in this paper.
This paper focuses on the development of a custom MEMS-based array which will facilitate cell secretion studies by enabling parallel electrochemical detection of secretion events from separate cells with millisecond resolution. Initial prototypes of the microarrays have been fabricated with well-shaped gold electrodes which roughly conform to the shape of a single cell. Amperometric measurements on bovine adrenal chromaffin cells using the prototype microarrays concluded that 80% of the catecholamine secreted from the cells was captured by the well-electrodes. This was a 4-fold increase in detection efficiency over the conventional carbon fiber electrode method. To expand the applicability of this method, additional cell-lines and microarray designs are under investigation. An amphibian fibroblast cell-line (FT cell-line, American Tissue Culture Collection) is being used in our lab. FT cells can take up hormones or other biological compounds from the culture media through a non-specific uptake mechanism which is still under investigation. Microarrays of a new design have been fabricated with patterned gold electrodes on polyimide. A different testing method will be applied to these new microarrays. The FT cells will be cultured directly on top of the microarrays to cover the gold electrodes. Cells will then be loaded with norepinephrine by incubation in media containing 1mM norepinephrine. Rapid elevation of intracellular Ca2+ levels triggers the exocytosis of norepinephrine which then can be detected by the gold electrode. The new polyimide based microarrays have been successfully used to support confluent growth of the FT cells. Loading of the FT cells with norepinephrine and electrochemical detection tests are underway.
We are focusing on the development of a biochip which will enable massively parallel amperometric measurements on single cells for exocytosis studies. Initial prototypes have been fabricated with picoliter-sized wells which roughly conform to the shape of the cells. The electrochemical measurement using the prototype devices can capture a large fraction, approximately 80%, of the catecholamine release with millisecond temporal resolution. With this prototype device, cells must be manually positioned into the micro-wells by a micromanipulator. Therefore, two new designs incorporating three dimensional microfluidic structures have been developed for automatic cell positioning. One design is based on thin silicon diaphragms with picoliter-sized well arrays, while another has 1μm silicon nitride membranes. Both designs have through-membrane holes and are designed in such a way that the cells will be automatically positioned onto electrodes once a suitable pressure differential is applied between the two sides of the thin diaphragms. Details of the microfabrication process for both designs will be presented in this paper as well as results of automatic cell positioning tests.
We are developing a novel readout for secretion of hormones and neurotransmitter on micro/nanofabricated chips. Traditional biochemical assays of signaling molecules secreted from cells are slow, cumbersome and have at best, a temporal resolution of several seconds. On the other hand, electrochemical measurement of hormone or transmitter secretion can obtain millisecond temporal resolution if the diffusion distance between the release site on the cell and the working electrode is within 1 micron. Carbon fiber microelectrodes can have millisecond time resolution, but can only measure release form a small fraction of the cell surface. We have fabricated arrays of Au electrodes in wells micromachined on the surface of silicon microchips. Each well/microelectrode roughly conforms to the shape of a single cell in order to capture release forma large fraction of the surface area of each cell with minimal diffusional delays. This paper will present details of the microfabrication process flow as well a initial results demonstrating millisecond-resolution measurement of catecholamine secretion form adrenal chromaffin cells. Our goal for this project is to develop enabling technology for massively parallel systems on a chip such as cell-based biosensors to detect neurotoxins and high-throughput assays of drugs that affect neurotransmitter release.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.