Biomedical Engineering – From Theory to Applications

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288 Biomedical Engineering – From Theory to Applications Fig. 12. Optical images of previously injected embryos showing development to the blastocyst stage. 11 Survival rates during the in-vitro development of embryos previously manipulated with pipette number 3 were similar to survival rates of control, non-manipulated embryos (Table 2). All 14 embryos pierced with pipette number 3 were monitored for in-vitro development, noting the survival rates at the two-cell, four-cell, morula, and blastocyst stages. Most micromanipulated embryos successfully developed to the two- and four-cell stages as well as at morula. The survival rate at all these stages was constant at 93%. Similarly, control embryos maintained a constant 87% survival rate at the same intervals. Only one of the 14 micromanipulated embryos remained arrested at the one-cell stage. This phenomenon was also observed in control embryos, suggesting that causes inhibiting division might be unrelated to micromanipulation factors. Gallium ions (unintentionally implanted during pipette milling) did not appear to affect development of mouse embryos to the blastocyst stage. While more detailed analyses are needed to fully address biocompatibility, these preliminary results are encouraging and could ultimately lead to improved piercing and survival rates during cell handling. 3. Sub-cellular monitoring, drug-delivery and manipulation The prior section reviewed recent developments in technologies aiming at improving the funtionalities of celllular tools (mostly manipulators and injectors) and addressing the quality of finishes in injection pipettes. In the spirit of completeness, in this section we will describe in a cursory manner, some of the current landscape for subcellular technology. 3.1 Monitoring and drug-delivery by sub-cellular smart particles The idea of targeting specific areas within the cell for detailed study is not new. Dyed biocompatible polymeric nanoparticles have been used for apoptosis imaging (Kim et al., 2006), and morphology of intracellular polymeric systems have been studied to address biocompatibility along with their suitability as imaging markers and as drug delivery carriers (Nishiyama, 2007). Carrier morphology studies suggest that particles of specific sizes are 11 With kind permission from Springer Science+Business Media: Biomedical Microdevices, Focus Ion Beam Micromachined Glass Pipettes for Cell Microinjection., Vol. 12, No.2, 2010, pp. (311–316), Campo, E.M., Lopez-Martinez, M. J., Fernández, E., Barrios, L., Ibáñez, E., Nogués, C., Esteve, J., & Plaza, J.A.
Micro-Nano Technologies for Cell Manipulation and Subcellular Monitoring required to prevent deleterious interaction with cell organelles. On those lines, vehicle morphology studies concluded that phagocitic cells responded differently to micelles (assemblies of hydrophobic/hydrophilic block-copolymers) of different sizes (Geng et al., 2007). Walter et al. examined polymeric spheres that were phagocited for drug delivery (Walter et al., 2001) and Akin et al. (Akin et al., 2007) used microbots (nanoparticles attached to bacteria) to deliver therapeutic cargo to specific sites within a cell. Microbots delivered nanoparticles of polystyrene carrying therapeutic cargo and DNA into cells by taking advantage of invasive properties of bacteria. Recently, Kataoka’s group (Mirakami et al., 2011) has sucessfully delivered chemotherapeutic drugs to the nuclear area of cancerous cells using micelles carriers. The specific delivery to the nuclear region is believed to have played a role in inhibiting the development of drug-resistance tumors. Within the subcellular domain, different approaches have aimed at manufacturing devices to interact with organelles. Some groups have contemplated the possibility of constructing micro total analysis stystems (µTAS) suitable for biological applications (Voldman et al., 1999), where the mechanisms to extract information out of the cellular entity are challenging. However, few attempts have been made to address viability and functionality of standard microtechnology processed systems. Recently, our group has reported silicon microparticles embedded in live cells, suggesting an outstanding compatibility between conventional microtechnology devices and live systems down to the cellular level (Fernandez-Rosas et al., 2009; Gomez-Martinez et al., 2010). In terms of sensing, initial functionality mechanisms have identified apoptosis. These revolutionary findings constitute a paramount paradigm shift on cellular metrology, histology, and drug delivery; which are likely to have a profound impact in future research lines. 3.2 Manipulation by biomimetics Another approach to sub-cellular exploration is inspired by nature. Indeed, understanding, mimicking, and adapting cellular and molecular mechanisms of biological motors in vitro has been forecast to produce a revolution in molecular manufacturing (Dinu et al., 2007, and Iyer et al., 2004). Biomolecular motors are biological machines that convert several forms of energy into mechanical energy. During a special session at Nanotech 2004 in Boston, MA, DARPA commissioned-overview by Iyer argued that functions carried out within a cell by biomolecular motors could be similar to man-made motors (i.e. load carrying or rotational movement). Researchers have already pondered about ways to transport designated cargo, such as vesicles, RNA or viruses to predetermined locations within the cell (Hess et al., 2008). Professor Hess during his keynote lecture at SPIE Photonics West ( January 2008) also proposed biomolecular motors as imaging and sensing devices. Biomolecular motors such as the motor protein kinesin have been suggested as efficient tractor trailers within the cell. Efficiency of these systems could generate useful tools (conveyor belts and forklifts) as nanoscale bio-manufacturing tools. Kinesin moves along a track and is responsible for transporting cellular cargo such as organelles and signaling molecules. However, a detailed explanation of this walking mechanism is still missing (Iyer et al., 2004), currently inhibiting spatial and temporal control of kinesin molecular motors. 3.3 Monitoring and manipulation by FIB and microfluidics Trends to intracellular manipulation also revolve around scaling down conventional pipettes. This trend is facilitated by microfluidics. Microsystem technologies have produced in the last decade an array of microfluidic devices (Verpoorte & De Rooig, 2003) that could 289
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BIOMEDICAL ENGINEERING - FROM THEOR
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VI Contents Chapter 9 Targeted Magn
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X Preface stand-alone and readers c
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120 6. Conclusion Biomedical Engine
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150 5. Conclusions Biomedical Engin
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162 1 st phase : half year 4 2 nd p
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166 7. Innovative active training B
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172 12. Maturing projects’ story
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180 16. Acknowledgments Biomedical
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190 R* M M M M MR*M O O O O M O R*
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196 Fig. 9. Chemical structure of 5
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198 Relative Intensity (AU) Biomedi
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204 2. Preparation of IO nanopartic
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454 In this case, the eigenvalues a
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456 where Biomedical Engineering -
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472 Nb b b l l1 Biomedical Engineer
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