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- Pdf Microdevices In Biology And Medicine (Artech House Methods In Bioengineering)
- Pdf Microdevices In Biology And Medicine Artech House Methods In Bioengineering
Bio-MEMS is an abbreviation for biomedical or biological microelectromechanical systems.
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Pdf Microdevices In Biology And Medicine (Artech House Methods In Bioengineering)
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Start by pressing the button below! Yarmush, M. Robert S. Langer, Sc. Zahn and Luke P. Library of Congress. Printed and bound in the United States of America. No part of this book may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, or by any information storage and retrieval system, without permission in writing from the publisher. All terms mentioned in this book that are known to be trademarks or service marks have been appropriately capitalized.
Artech House cannot attest to the accuracy of this information. Use of a term in this book should not be regarded as affecting the validity of any trademark or service mark. Hiding under the shiny coat of our cars, iPods, cellular phones, laptops, and televisions, the integrated circuit and silicon microchip have changed the way we live forever.
Features a thousand times smaller than a single millimeter enable an unparallel control over electrical signals resulting in nearly magical computational, communication, and memory powers. At the dawn of the twenty-first century, a similar revolution is changing the study of biology and the practice of medicine. Microscale patterns, three-dimensional features, and the physics of small places offer to radically change our ability to screen thousands of conditions, control the cellular microenvironment, and provide innovative tools for the diagnosis and treatment of disease.
Notably, microdevices that have already reached the market are gaining increasing popularity. Perhaps the most celebrated application of microtechnology is the Affymetrix GeneChip, a DNA microarray capable of screening the relative transcription of tens of thousands of genes, essentially the entire genome, in a single experiment.
First published in by the Patrick O. Brown group at Stanford University, the microarray spotting approach has spawned many variants such as chromatin immuneprecipitation on chip ChIP-on-chip and SNP profiling. The GeneChip microarray has become a standard tool for the screening of complex genetic information. Another commercially available system that is rapidly growing in popularity is the Agilent Bioanalyzer, a microfluidics-based microchip that uses electrophoresis for the separation of RNA, DNA, and proteins.
The newest models allow for on-chip staining and flow cytometry analysis of a small number of cells by replacing electrophoresis with a pressure-driven flow. Finally, PillCam is a commercially available microdevice that conjures up visions of the film The Fantastic Voyage. Developed by Given Imaging, PillCam is a capsule measuring 11 by 26 mm and weighing less than 4 grams.
It contains a miniaturized imaging device that takes up to 14 images per second as it passes down the gastrointestinal tract. The success of these early microdevices has brought us to realize the need for a methods-based book that will provide timely insight into the technology of newly developed bio-MEMS devices. Methods in Bioengineering: Microdevices in Biology and Medicine is intended for students and scientists who wish to apply these tools for basic science or clinical diagnostics and for clinicians who wish to familiarize themselves with the science of this emerging technology.
As part of the Artech House Methods in Bioengineer- xi Preface ing Series, this book presents the science behind microscale device design as well as the engineering of its fabrication.
Each chapter includes a detailed, step-by-step methodology as well as a troubleshooting table designed to enable the rapid dissemination of microfabrication technology. Supported by dozens of full-color illustrations, this book covers the microfabrication technology involved in developing microdevices for biological applications and from bench to bedside.
Readers will gain a unique perspective on the challenges and emerging opportunities in developing microdevices for cell capture from whole blood, study of transcriptional dynamics in living cells, temporal control of cell-cell interactions, nanoscale measurements of cellular forces, immobilization of living organisms, optical and electrical on-chip cell sorting, human-on-chip models of drug metabolism, microreactors for tissue engineering, and 3-D control of the cellular microenvironment.
We hope you enjoy this book as much as we enjoyed putting it together. Yaakov Nahmias, Ph. Bhatia, M. Isolating purified, homogeneous cells from complex biological samples, however, is a lengthy process suited for specialized, well-equipped research laboratories and difficult to implement in clinical medicine.
Here we outline a rapid and easy process for utilizing state-of-the-art microfluidic technology to isolate leukocyte subpopulations directly from whole blood using neutrophils as an example. Cellular phenotype is also of interest in immunology, where researchers seek to understand the immune system through its individual cellular and molecular components.
Standard techniques exist for cellular enumeration and cellular fractionation from complex samples, including density gradient centrifugation, negative selection techniques such as RosetteSep, and positive selection techniques such as fluorescence-activated cell sorting FACS and magnetic-activated cell sorting MACS.
These different methods typically require highly trained technical staff processing samples over a period of hours. In the case of FACS and centrifugation techniques, they require large, specialized equipment as well.
Our lab is interested in building tools that enable clinicians to rapidly and easily study the immune system to help doctors predict clinical outcomes in patients. We have developed a set of tools that rapidly and efficiently captures cells on the walls of microfluidic devices using antibody-affinity isolation . The well-defined fluid flow in microfluidic channels translates into precise shear forces seen by cells near the surfaces of the microfluidic device.
This, combined with the specificity of monoclonal antibodies for cell-surface antigens, leads to highly specific capture of cells in these devices [4, 5]. A given cell type with a specific cell-surface antigen can thus be isolated by designing a microfluidic device, coating it with a specific antibody, and flowing the biological sample through the device at a specific flow rate.
This protocol outlines the overall process for the design, manufacture, and testing of devices for rapid, specific isolation of granulocytes directly from whole blood. Granulocytes are a particularly challenging cell to process using standard density gradient techniques because they are easily activated by sample processing and because they are short-lived [6, 7]. The design shown in Figure 1. The design maximizes the total device width across the long dimension of a standard microscope slide in order to maximize the flow rate for a given shear stress and a given height, thus minimizing processing time.
Device operation consists of flowing the blood through the device for 5 minutes, washing the device with physiological saline buffer for 5 minutes, and then processing the captured cells for downstream analysis. The devices are made through standard PDMS rapid-prototyping methods . PDMS is a flexible elastomer that can be chemically modified with biomolecules. This protocol outlines the process by which PDMS devices are fabricated, coated with antibodies, and used to capture and process cells.
The design used here has been used by engineers, scientists, and technicians with equal success. While the protocol here describes capture of granulocytes, we have adapted it for capture of T and B lymphocytes, monocytes, and other rarer populations found in circulating blood. The main design considerations therefore are the particular antibody-antigen pair that will be used to capture cells and the device geometry and fluid flow conditions that give the proper shear forces at the surface of the interaction.
The optimal antibody-antigen interaction is typically determined by consulting standard resources that list cell-surface antigens as well as their distribution on the cell surface . Once a set of suitable antibodies has been identified, typically one can validate the presence and uniqueness of the interaction on a flow cytometer. A discussion of flow cytometry is outside the scope of this protocol, and the reader is referred to many excellent reviews on the subject .
The next design parameter is the optimal shear stress for cell capture for a given cell and antibody. While specific microfluidic devices have been designed to determine optimal flow conditions for capture [3, 11, 12], generally it is straightforward to run multiple devices with a design as given in Figure 1.
Capture is assessed for purity, total number, and efficiency at each condition and design changes are made to meet target requirements.
The design in Figure 1. Efficiency generally can be increased by decreasing the distance between the parallel plates data not shown , increasing the length of the channel , or adding obstacles to increase surface area and break up flow streamlines along the length of the device . Purity is mainly determined by the uniqueness of a specific antibody-antigen pair. Any experiment involving cells or human samples adds additional variables to experimental design.
Samples that contain cells can be heterogeneous in surface marker expression, and cell expression of surface antigens can change over time. Antibody cap3 Immunoaffinity Capture of Cells from Whole Blood ture of cells can cause crosslinking of proteins, which can lead to activation of cell-signaling pathways that affects cellular phenotype.
Clinical samples can be extremely variable in cell numbers, activation states, and antigen expression. For small proof-of-concept studies, 6 to 10 subjects are usually sampled, with multiple devices used for different downstream analysis.
Despite complications in cellular materials, the microfluidic capture described in this protocol is extremely robust and used by many researchers without formal training in microfluidics. The last set of design parameters is determined by the downstream application of captured cells. For genomic studies, it is of utmost importance to maintain nuclease-free conditions while preparing and processing samples. For proteomics, it is necessary to assess the design in terms of material compatibility with downstream processes.
Mass spectrometry—based methods are particularly prone to chemical contaminants and protein background. Furthermore, for either method, it is helpful to develop a specification for total protein or nucleic acid that is needed for downstream analysis. The device is then scaled to capture sufficient numbers of cells to meet the processing specification. These devices are then chemically coated with antibodies to capture cells from biological fluids.
The reagents, materials and common supplies, and equipment necessary for these processes are outlined below in Tables 1. Many laboratories have devised unique solutions to interfacing microfluidic devices with macroscopic fluid-handling devices pumps, valves, and so forth. Figure 1. Table 1. Device fabrication can be divided into three main parts: 1 generation of a master of SU-8 on a silicon wafer; 2 replicating the part with a flexible elastomer, including demolding, sectioning, and cutting fluidic interconnect ports; and 3 device bonding to a PDMS or glass substrate.
The injection syringe is a standard 1 mL syringe with a 22G SS blunt-tip needle blue. The Teflon outlet tubing is made from a 0. Device features are photocrosslinked onto the silicon wafer with a UV lamp and a transparency mask containing the device. The generation of a master with SU-8 on a silicon wafer typically requires special facilities Class clean room or contracting from outside vendors.
The process by which a master is generated is reviewed elsewhere in this book see Chapter 2 or Chapter 5 and will not be repeated here. Once a completed master is obtained, it is ready to replicate with PDMS. SUon-silicon masters, if handled carefully, typically last for more than molding cycles, as described below.
The following molding procedures in our lab are carried out in a large, class , clean room environment in order to minimize defects caused by environmental particles. It is mixed in a container, poured over the master, degassed, and cured overnight. Once cured, it can be peeled off the master, creating a negative cast of the master.
The process is outlined next. Sections highlighted in yellow are covered in this section. Put on a clean pair of gloves, lab coat, and face mask. Remove the silicon master from its protective case and place it in a petri dish secured by tape, with SU-8 features facing upwards.
Pdf Microdevices In Biology And Medicine Artech House Methods In Bioengineering
This content was uploaded by our users and we assume good faith they have the permission to share this book. If you own the copyright to this book and it is wrongfully on our website, we offer a simple DMCA procedure to remove your content from our site. Start by pressing the button below! Yarmush, M. Robert S.
BioMEMS devices are as important to the future of medicine as microprocessors were to the computer revolution at the end of the last century. BioMEMS is a science that includes more than simply finding biomedical applications for microelectromechanical systems devices. It represents an expansion into a host of new polymer materials, microfluidic physics, surface chemistries and their modification, "soft" fabrication techniques, biocompatibility, and cost-effective solutions to biomedical problems. It brings together the creative talents of electrical, mechanical, optical, and chemical engineers, materials specialists, clinical laboratory scientists, and physicians. BioMEMS devices are the platform upon which nanomedicine will be delivered. Based on the author's course on bioMEMS at the University of Minnesota, this book is an introduction to the science and a survey of the state of the art.
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The following pages represent in a much extended form the lectures and laboratory exercises given by the author before his botanic classes at the university of pennsylvania, and before public audiences else where, especially, farmers institutes with which he has had three years experience as a lecturer in pennsylvania. Fundamentals of biomems and medical microdevices on. Fundamentals of biomems and medical microdevices Periodic flowstop for mammalian and human embryonic stem. Version details trove remote sensing from air and space r.
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