Pulsed Laser Microbeams for Cellular Manipulation

Pulsed Laser Microbeams for Cellular Manipulation

Author: Amy Noel Stacy Hellman

Publisher: ProQuest

Published: 2008

Total Pages: 174

ISBN-13: 9780549605348

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Highly focused pulsed laser radiation (pulsed laser microbeams) provide the ability to deposit energy with high spatial precision and controllable cellular damage. As a result, pulsed laser microbeams have been explored as a fast, non-contact means for cellular manipulations such as cellular microsurgery, transient cell membrane permeabilization, and targeted cell lysis. In this dissertation we examine the mechanisms of highly focused laser microbeams of nanosecond and picosecond duration to achieve cell lysis, cell necrosis, and molecular delivery. We have developed a time-resolved imaging system to visualize these processes with nanosecond temporal resolution and use image analysis to measure the physical perturbation applied to the cells as a function of laser microbeam pulse energy and pulse duration. Fluorescence assays are used to assess the biological response (necrosis, molecular delivery, and biochemical pathway activity) to the laser microbeam irradiation, and a biophysical model is developed to establish connections between specific physical characteristics and the resulting cellular effect. Our studies reveal that pulsed laser microbeam processes are mediated by optical breakdown resulting in plasma formation, shock wave emission, and cavitation bubble formation, expansion, and collapse. Cavitation bubble expansion was found to be the primary mechanism responsible for cellular modification. Hydrodynamic analysis based on the measured time evolution of the cavitation bubble growth, combined with assessment of the cellular response, revealed that the maximum wall shear stress associated with the cavitation bubble expansion governs the location and spatial extent of cell lysis, cell necrosis, and molecular delivery. In addition, we demonstrate how the variation of laser microbeam pulse duration can allow modulation of the spatial extent of cellular modification in order to tailor the cellular perturbations and optimize specific applications. These detailed studies provide a basis for the informed selection of specific laser parameters (i.e. pulse duration and energy) to achieve a desired cellular outcome, with controllable degrees of collateral damage. We demonstrate three different applications in which the laser parameters are well matched for particular applications in cellular analytics, activation of biochemical pathways, and microfluidics.


Laser Manipulation of Cells and Tissues

Laser Manipulation of Cells and Tissues

Author: Michael W. Berns

Publisher: Academic Press

Published: 2007-06-20

Total Pages: 824

ISBN-13:

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The use of laser-based manipulation tools has literally exploded on the cell biology and molecular pathology scene, resulting in as many different laser micromanipulation systems as there are people using them. Laser Manipulation of Cells and Tissues ties all these systems and studies together, describing all of the different kinds of research and practical/analytical applications of laser manipulation. It also provides the reader with the basic information needed to actually build one's own laser micro-manipulation system. The combination of imaging and molecular probe technology with laser micromanipulation greatly extends the use of this technology in molecular, cellular, developmental and patho-biology/medicine. This book should be valuable to scientists, clinicians, and students in the fields of cell and developmental biology, cell physiology, cancer biology, pathology, and stem cell biology. Devotes four chapters to laser catapulting and capture of DNA and other cellular material for biochemical analysis - a major use of this technology that has been adapted for molecular pathology both in clinical medicine and research Discusses the theory of laser tweezers (optical tweezers) and its application to novel problems in biology Covers topics on optoporation (getting things into cells), uncaging of molecules, and the ability to collect and analyze nanomolar amounts of cell material by an array of biochemical/physical tools of particular interest to cell biologists and drug discovery researchers


Micromanipulation by Light in Biology and Medicine

Micromanipulation by Light in Biology and Medicine

Author: Karl Otto Greulich

Publisher: Springer Science & Business Media

Published: 2012-12-06

Total Pages: 308

ISBN-13: 1461241103

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There are probably few people who do not dream of the good old times, when do ing science often meant fascination, excitement, even adventure. In our time, do ing science involves often technology and, perhaps, even business. But there are still niches where curiosity and fascination have their place. The subject of this book, technological as its title may sound, is one of the fortunate examples. It will report on lasers generating the coldest places in the Universe, and on table top laser microtools which can produce a heat "inferno" as it prevails in the interior of the Sun, or simulate, for specific plant cells, microgravity of the space around our plan et Earth. There will be some real surprises for the reader. The applications range from basic studies of the driving forces of cell division (and thus life) via genetic modification of cells (for example, for plant breeding) to medical applications such as blood cell analysis and finally in vitro fertilization. What are these instruments: laser microbeams and optical tweezers? Both are lasers coupled with a fluorescence microscope. The laser microbeam uses a pulsed ultraviolet laser. Light is focused, as well as possible, in space and time, in order to obtain extremely high light intensities - high enough to generate, for a very short instant, extremely hot spots which can be used to cut, fuse or perforate biological material.


Microtechnology for Cell Manipulation and Sorting

Microtechnology for Cell Manipulation and Sorting

Author: Wonhee Lee

Publisher: Springer

Published: 2016-10-05

Total Pages: 287

ISBN-13: 3319441396

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This book delves into the recent developments in the microscale and microfluidic technologies that allow manipulation at the single and cell aggregate level. Expert authors review the dominant mechanisms that manipulate and sort biological structures, making this a state-of-the-art overview of conventional cell sorting techniques, the principles of microfluidics, and of microfluidic devices. All chapters highlight the benefits and drawbacks of each technique they discuss, which include magnetic, electrical, optical, acoustic, gravity/sedimentation, inertial, deformability, and aqueous two-phase systems as the dominant mechanisms utilized by microfluidic devices to handle biological samples. Each chapter explains the physics of the mechanism at work, and reviews common geometries and devices to help readers decide the type of style of device required for various applications. This book is appropriate for graduate-level biomedical engineering and analytical chemistry students, as well as engineers and scientists working in the biotechnology industry.


Laser Imaging and Manipulation in Cell Biology

Laser Imaging and Manipulation in Cell Biology

Author: Francesco S. Pavone

Publisher: John Wiley & Sons

Published: 2011-07-28

Total Pages: 229

ISBN-13: 3527632255

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Here, the editor has gathered a team of international experts to present the latest advances in the field of laser imaging and manipulation techniques. The result is broad coverage of the interactions with biological samples to perform novel optical manipulation operations, both on the cellular and tissue levels. Of interest to physicists working and researching laser tissue mechanisms, cell biologists investigating new imaging and manipulation operation on the cellular level, medical doctors working with new laser therapies and diagnostic tools, as well as engineers developing new technologies in the field of optics and lasers.


Intelligent Robotics and Applications

Intelligent Robotics and Applications

Author: Honghai Liu

Publisher: Springer Nature

Published: 2022-08-03

Total Pages: 813

ISBN-13: 3031138228

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The 4-volume set LNAI 13455 - 13458 constitutes the proceedings of the 15th International Conference on Intelligent Robotics and Applications, ICIRA 2022, which took place in Harbin China, during August 2022. The 284 papers included in these proceedings were carefully reviewed and selected from 442 submissions. They were organized in topical sections as follows: Robotics, Mechatronics, Applications, Robotic Machining, Medical Engineering, Soft and Hybrid Robots, Human-robot Collaboration, Machine Intelligence, and Human Robot Interaction.


Microtubules: in vivo

Microtubules: in vivo

Author:

Publisher: Academic Press

Published: 2010-09-24

Total Pages: 549

ISBN-13: 0123813506

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Microtubules: in vivo includes chapters by experts around the world on many aspects of microtubule imaging in living and fixed cells; assays to study microtubule function in a wide array of model organisms and cultured cells; high resolution approaches to study of the cytoskeleton. The authors share their years of experience, outlining potential pitfalls and critical factors to consider in experimental design, experimental implementation and data interpretation. Includes chapters by experts around the world on many aspects of microtubule imaging in living and fixed cells; assays to study microtubule function in a wide array of model organisms and cultured cells; high resolution approaches to study of the cytoskeleton The authors share their years of experience, outlining potential pitfalls and critical factors to consider in experimental design, experimental implementation and data interpretation