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Lumped Parameters Model 2. Worked Example 2—Plasma Extraction from Blood 2. Establishment of the Flow 2.
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Lumped Parameters Model 2. Worked Example 2—Plasma Extraction from Blood 2. Establishment of the Flow 2. Dean Flow 2. Helle-Shaw Flows 2. Droplet Engulfment 4. Concentration 5. Diffusion or Sedimentation 5. Drug Diffusion in the Human Body 5. Light Scattering 8. Electrophoresis Starting from the very beginning of this science in the s, spectacular advances have been made, such as the completion of the determination of the human genome sequence, and dramatic changes have broken out in the field of proteomics and, more recently, new breakthroughs have been made in the domain of cellular analysis.

The field of investigations of biotechnology has constantly increased, from the first biochips built to analyze sequences of DNA and investigate its mutations to protein analysis and the study of role of proteins in the human life and the comprehension of the complex mechanisms that take place inside the cells. Biotechnology is a science that is not only dedicated to assist biologists in their desire to understand the complexity of life. It also has very practical applications, especially in bioanalysis and biodetection.

Biotechnology is not restricted to in vitro analysis, but has direct implications for in vivo treatments. Concerning the in vivo domain, the impact of the new technologies is manifold. First, the monitoring of the correct functioning of some vital organs in patients at risk is going to be possible.

Second, miniaturization techniques will greatly reduce the invasiveness of external interventions inside the human body. Third, new biotechnological devices may help internal drug guidance to find their targets inside the human body. Fourth, new cell encapsulation techniques are going to improve considerably the human organs grafting. Finally, a trend that many of us will experience in the years to come is towards automated medical help and monitoring right at home.

Biotechnological microsystems are called different names having more or less the same physical meaning, such as biochips, or bioMEMS—for microelectromechanical systems, or lab-on-a-chip—meaning that many of the different operations performed in a lab are done on a single microdevice and sometimes mTAS micrototal analysis systems. It is surprising how the concept and development of the first DNA biochip opened the way to a completely new domain of technology. It soon appeared that many other concepts could be imagined and that miniaturization had many advantages in biochemical science.

A first advantage resides in the automation and streamlining of biological processes, as shown with the DNA biochip: Another example is that of the proteomic reactors breaking proteins into peptides by enzymatic catalyst inside microchannels; the peptides are then transported by a buffer fluid to a spray injector and then to a mass spectrometer where the peptides are identified.

Such a protein chip realizes many operations in sequence which otherwise would have needed many different manipulations and a lot of time. Another advantage of biotechnological microdevices is the reduction of costs of biological analysis due not only to the streamlining and parallelization of the operations, but also to the reduction of the quantities of reactants. Because the reactants needed to perform the sequence of biological reactions are usually quite expensive, it is important that they be used in very limited quantities.

Of course, biochips may still be somewhat expensive, especially if etched silicon is used, but it is more than compensated by the gain in the mass of reactants. Explosive substances are not dangerous anymore at very low concentrations, and dangerous viruses and toxic bacteria can be more easily confined in microsystems. It is also expected that biochips or bioMEMS can provide higher sensitivity than usual macroscopic systems.

For example, some diseases caused by a virus can be detected earlier, at a number of viruses much smaller than the usual diagnostics, leading to better treatment and a reduction of the contagion possibility. On the research point of view, there are also many advantages brought by biotechnological microsystems.

For example, in vivo interventions are facilitated by the small size of the new biotechnological devices, reducing the invasiveness of the drug delivery system. Another example is the technology of encapsulated active microparticles targeted specifically in the human body. It is expected that biochips will also contribute to the discovery of new drugs, by testing many new molecules at the same time on living cells isolated in lab-on-a-chip for cells.

In that sense, biotechnology is increasingly considered a very useful complement to biology itself, as it may contribute to discover new drugs by automatically testing many molecules at the same time.

As mentioned with the complementarity between biotechnology and biology, we point out here that the central theme of biotechnology is the control, displacement, and guidance of the different micro-sized objects that are present in a biologic buffer liquid. In reality, there are three types of biological objects. Their sizes range from about 20 nm short strands of DNA to mm for the larger cells.

In this category, we can list magnetic beads, different fluorophores CY3, CY5, FITC , quantum dots, gold microparticles, polypyrolles, carbon nanowires, and surfactants. These objects are generally smaller than the previous ones, ranging from 10 nm to 2 mm. Recently, a third type Preface xiii of biological objects has appeared: These objects are new composites—like polymeric or gelled capsules containing cells, bacteria, or proteins—and they bear great hopes in medical treatments; their size can vary between a few microns and mm.

We will refer to all these objects under the names of micro- and nanoparticles and macromolecules. A simplified statement is that biologists study the functional and chemical behavior of biological objects, whereas biotechnologists focus on the mechanical and chemical behavior of these objects.

A book on a subject as rapidly evolving as microfluidics for biotechnology necessarily reflects the state of the art at a certain time. In the first edition of this book in , the focus was on microfluidics for lab-on-chips dedicated to DNA analysis and immunoassays for protemics. Since that time, cellomics has seen considerable developments. Many efforts have been dedicated to the study of cells, which is the key to understanding the functioning of the complex human system and to the development of new drugs.

Attention has been given to single-cell studies and communication between cells. Hence, transport and manipulation of cells have become an important topic. In the wake of the development of cellular mechanics and cellular microfluidics, triggered by cell transport and encapsulation applications, digital and especially droplet microfluidics have seen a considerable boost.

Another recent evolution is the development of the use of biological liquids—whole blood and alginates, for example—in in vitro biochips. This evolution has promoted the study of the rheology of biopolymers and their non-Newtonian, viscoelastic behavior in microsystems. This second edition reflects this evolution and incorporates new concepts in cellular microfluidics and cell manipulation, along with rheological considerations on viscoelastic liquids.

A new chapter devoted to digital and droplet microfluidics has been introduced. However, it has seemed important to the authors to keep the theoretical fluid mechanics basis in order to maintain the coherence of the text and to provide a stand-alone book.

Hence, this new edition has globally conserved the frame of the first edition. Physical laws do not change between macroscopic and microscopic scales, but the relative importance of the different forces is considerably changed between these two scales. In Chapter 1, the scaling of the different forces as a function of the dimension of the system is presented and the dominating forces and phenomena are pointed out.

Nondimensionless numbers and characteristic times of the different phenomena associated to microfluidics are presented and discussed. Chapters 2, 3, and 4 are dedicated to the microfluidics aspects of the buffer fluid in biochips. In order to predict correctly the behavior of particles, the physical behavior of the buffer carrier fluid must be first determined. Chapter 2 treats continuous single-phase microflows.

Theoretical bases are given first. A section dedicated to the rheology of non-Newtonian fluids in biotechnology has been incorporated, taking the example of alginate solutions, which are now widely used. Because of the growing importance of cell separation devices, emphasis has been placed on microfluidic networks.

For this reason, it has been found that a chapter dedicated to the notions of interface, capillarity, and static droplet behavior was needed. This is the object of Chapter 3. To go further in the investigations of new multiphase microfluidic solutions, Chapter 4, devoted to digital and droplet microfluidics, has been added to this second edition.

These new techniques appear to be promising ways of transporting biological objects in extremely small liquid volumes. In digital microfluidic applications, microdrops of a few tenths of micrometers are moved individually step by step on a flat surface. In droplet microfluidic applications, same liquid volumes are transported within an immiscible continuous microflow. Because the micro-and nanoparticles and macromolecules in which we are interested are much larger than the fluid molecules, their behavior differs from that of the fluid.

Therefore, Chapters 5 and 6 focus on the mechanical behavior of the particles themselves, under the action of diffusion Chapter 5 and transport by advection Chapter 6.

Different numerical approaches are presented, such as continuum-based numerical and discrete methods. A special addition concerning cellular microfluidics i. All the studies on the buffer carrier fluid flow and the behavior of the particles in this flow are aimed at controlling the motion of the particles of interest to have them placed at some specific location in order to be able to perform the desired reaction or analysis.

Chapter 7 is dedicated to the study of biochemical reactions. First, the principle of biorecognition is presented and the different biochemical reactions to recognize DNA sequences and antibodies are studied. It has been found useful to precisely determine the nature and the characteristic of the most used targets in biotechnology biological targets and synthetic particles.

Chapter 8 describes the characteristics of these particles and introduces an experimental aspect by presenting the methods used to manipulate or characterize them. Because the transport by the buffer fluid is often not specific enough, complementary methods have been developed. In Chapter 9, we present the principle of labeled magnetic microbeads and show how these beads are used to bind with the targeted biological objects and to transport them into specifically designated areas.

Another usual way of controlling the motion of microparticles is based on the use of electric fields. In Chapter 10, we present the different ways electric fields act on the particles, like electrophoresis and dielectrophoresis. We finally conclude in Chapter 11 by recalling the main recent developments and the future trends.

We thank our colleagues, particularly N. Sarrut, for their contribution with photographs, and A. Buguin who has been kind enough to review and comment on some chapters. Our discussions with A. Ajdari, R. Austin, D. Chatenay, J. Joanny, F.

Perraut, J.

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Microfluidics for Biotechnology, Second Edition

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