Ranjeet Kumar; Chandra Shakher; Dalip Singh Mehta
We demonstrate a simple and compact optical interferometric unit combined with a conventional optical tweezer system for simultaneous multiple trapping and micromanipulation of mono-dispersed polystyrene spheres and aggregation of small-floating clusters of single-walled carbon nanotubes (SWCNTs). The interferometric unit was made compact by means of coating a thin layer of aluminum oxide on one side of the cubic beam splitter (CBS) which works as a static reference mirror and an adjustable mirror was mounted on a XYZ translational stage facing the other adjacent side of the CBS. Thus, the developed interferometric unit is quite analogous to a Michelson interferometer but is compact. Sinusoidal interference fringes with variable carrier frequency and orientations were generated. The interference fringes were then used for multiple trapping of polystyrene spheres along bright fringes resulting in pattern formation and also the aggregation of tiny floating clusters of SWCNTs. The proposed system is compact and easy to align because of its common-path geometry.
DOI
Friday, August 6, 2010
Thursday, August 5, 2010
Biophotonic techniques for the study of malaria-infected red blood cells
Jakob M. A. Mauritz, Alessandro Esposito, Teresa Tiffert, Jeremy N. Skepper, Alice Warley, Young-Zoon Yoon, Pietro Cicuta, Virgilio L. Lew, Jochen R. Guck and Clemens F. Kaminski
Investigation of the homeostasis of red blood cells upon infection by Plasmodium falciparum poses complex experimental challenges. Changes in red cell shape, volume, protein, and ion balance are difficult to quantify. In this article, we review a wide range of optical techniques for quantitative measurements of critical homeostatic parameters in malaria-infected red blood cells. Fluorescence lifetime imaging and tomographic phase microscopy, quantitative deconvolution microscopy, and X-ray microanalysis, are used to measure haemoglobin concentration, cell volume, and ion contents. Atomic force microscopy is briefly reviewed in the context of these optical methodologies. We also describe how optical tweezers and optical stretchers can be usefully applied to empower basic malaria research to yield diagnostic information on cell compliance changes upon malaria infection. The combined application of these techniques sheds new light on the detailed mechanisms of malaria infection providing potential for new diagnostic or therapeutic approaches.
DOI
Investigation of the homeostasis of red blood cells upon infection by Plasmodium falciparum poses complex experimental challenges. Changes in red cell shape, volume, protein, and ion balance are difficult to quantify. In this article, we review a wide range of optical techniques for quantitative measurements of critical homeostatic parameters in malaria-infected red blood cells. Fluorescence lifetime imaging and tomographic phase microscopy, quantitative deconvolution microscopy, and X-ray microanalysis, are used to measure haemoglobin concentration, cell volume, and ion contents. Atomic force microscopy is briefly reviewed in the context of these optical methodologies. We also describe how optical tweezers and optical stretchers can be usefully applied to empower basic malaria research to yield diagnostic information on cell compliance changes upon malaria infection. The combined application of these techniques sheds new light on the detailed mechanisms of malaria infection providing potential for new diagnostic or therapeutic approaches.
DOI
Probing oxidative stress in single erythrocytes with Raman Tweezers
E. Zachariah, A. Bankapur, C. Santhosh, M. Valiathan and D. Mathur
Raman Tweezers have been successfully applied to characterize chemically-induced oxidative stress on optically-trapped live, single erythrocytes. There is significant enhancement in Raman peak intensities corresponding to S=S and C–S stretching modes that are induced by oxidative stress. This is consistent with the formation of mixed disulphides between protein SH groups and low-molecular-mass thiols such as glutathione during oxidative damage to cells. Enhancement in glutathione level as a protective response against oxidative stress has been observed. Principal component analysis of the data yields good discrimination between spectra of normal and stress-induced red blood cells.
Raman Tweezers have been successfully applied to characterize chemically-induced oxidative stress on optically-trapped live, single erythrocytes. There is significant enhancement in Raman peak intensities corresponding to S=S and C–S stretching modes that are induced by oxidative stress. This is consistent with the formation of mixed disulphides between protein SH groups and low-molecular-mass thiols such as glutathione during oxidative damage to cells. Enhancement in glutathione level as a protective response against oxidative stress has been observed. Principal component analysis of the data yields good discrimination between spectra of normal and stress-induced red blood cells.
Tunable optical gradient trap by radial varying polarization Bessel-Gauss beam
Gao, X.-M., Hu, S., Li, J.-S., Ding, Z.-H., Guo, H.-M., Zhuang, S.-L.
Optical tweezers play an important role in many domains, especially in life science. And optical gradient force is necessary for constructing optical tweezers. In this paper, the optical gradient force in the focal region of radial varying polarization Bessel- Gauss beam is investigated numerically by means of vector diffraction theory. Results show that the beam parameter and vary rate parameter that indicates the change speed of polarization rotation angle affect the optical gradient force pattern very considerably, and some novel force distributions may come into being, such as multiple force minimums, force ring, and force crust. Therefore, the focusing of radial varying polarization Bessel-Gauss beam can be used to construct optical traps.
DOI
Optical tweezers play an important role in many domains, especially in life science. And optical gradient force is necessary for constructing optical tweezers. In this paper, the optical gradient force in the focal region of radial varying polarization Bessel- Gauss beam is investigated numerically by means of vector diffraction theory. Results show that the beam parameter and vary rate parameter that indicates the change speed of polarization rotation angle affect the optical gradient force pattern very considerably, and some novel force distributions may come into being, such as multiple force minimums, force ring, and force crust. Therefore, the focusing of radial varying polarization Bessel-Gauss beam can be used to construct optical traps.
DOI
Membrane nanotubes drawn by optical tweezers transmit electrical signals between mammalian cells over long distances
Pedro Pascoal, Davor Kosanic, Marinela Gjoni and Horst Vogel
Biological cells continuously change shape allowing essential functions such as cell motility, vesicle-mediated release/uptake of soluble and membrane components or nanotube-mediated cell–cell communications. Here we use single cell micromanipulation to induce functional changes of cell shape for nanobiotechnological applications. Optical tweezers are focused on the plasma membrane of living cells to pull membrane nanotubes of 200 nanometre diameters and 100 micrometre lengths. Upon switching off the laser tweezer membrane nanotubes relax back to the cell surface. Single-exponential relaxation times deliver local mechanical properties of cells' plasma membrane. Nanotubes pulled beyond 100 micrometre tear off and form micrometre-sized vesicles carrying functional membrane receptors and cytoplasmic signaling components. Membrane nanotubes from one cell can be contacted to adjacent cells forming via connexins intercellular electrical connections within seconds in all directions. Our method opens broad applications for multiplexing single-cell analytics to submicrometer/subfemtoliter ranges and for creating artificial intercellular signaling networks, both not attainable by current methodologies.
DOI
Biological cells continuously change shape allowing essential functions such as cell motility, vesicle-mediated release/uptake of soluble and membrane components or nanotube-mediated cell–cell communications. Here we use single cell micromanipulation to induce functional changes of cell shape for nanobiotechnological applications. Optical tweezers are focused on the plasma membrane of living cells to pull membrane nanotubes of 200 nanometre diameters and 100 micrometre lengths. Upon switching off the laser tweezer membrane nanotubes relax back to the cell surface. Single-exponential relaxation times deliver local mechanical properties of cells' plasma membrane. Nanotubes pulled beyond 100 micrometre tear off and form micrometre-sized vesicles carrying functional membrane receptors and cytoplasmic signaling components. Membrane nanotubes from one cell can be contacted to adjacent cells forming via connexins intercellular electrical connections within seconds in all directions. Our method opens broad applications for multiplexing single-cell analytics to submicrometer/subfemtoliter ranges and for creating artificial intercellular signaling networks, both not attainable by current methodologies.
DOI
TweezPal – Optical tweezers analysis and calibration software
Natan Osterman
Optical tweezers, a powerful tool for optical trapping, micromanipulation and force transduction, have in recent years become a standard technique commonly used in many research laboratories and university courses. Knowledge about the optical force acting on a trapped object can be gained only after a calibration procedure which has to be performed (by an expert) for each type of trapped objects. In this paper we present TweezPal, a user-friendly, standalone Windows software tool for optical tweezers analysis and calibration. Using TweezPal, the procedure can be performed in a matter of minutes even by non-expert users. The calibration is based on the Brownian motion of a particle trapped in a stationary optical trap, which is being monitored using video or photodiode detection. The particle trajectory is imported into the software which instantly calculates position histogram, trapping potential, stiffness and anisotropy.
DOI
Optical tweezers, a powerful tool for optical trapping, micromanipulation and force transduction, have in recent years become a standard technique commonly used in many research laboratories and university courses. Knowledge about the optical force acting on a trapped object can be gained only after a calibration procedure which has to be performed (by an expert) for each type of trapped objects. In this paper we present TweezPal, a user-friendly, standalone Windows software tool for optical tweezers analysis and calibration. Using TweezPal, the procedure can be performed in a matter of minutes even by non-expert users. The calibration is based on the Brownian motion of a particle trapped in a stationary optical trap, which is being monitored using video or photodiode detection. The particle trajectory is imported into the software which instantly calculates position histogram, trapping potential, stiffness and anisotropy.
DOI
Monitoring of laser micromanipulated optically trapped cells by digital holographic microscopy
Björn Kemper, Patrik Langehanenberg, Alexander Höink, Gert von Bally, Falk Wottowah, Stefan Schinkinger, Jochen Guck, Josef Käs, Ilona Bredebusch, Jürgen Schnekenburger, Karin Schütze
For a precise manipulation of particles and cells with laser light as well as for the understanding and the control of the underlying processes it is important to visualize and quantify the response of the specimens. Thus, we investigated if digital holographic microscopy (DHM) can be used in combination with microfluidics to observe optically trapped living cells in a minimally invasive fashion during laser micromanipulation. The obtained results demonstrate that DHM multi-focus phase contrast provides label-free quantitative monitoring of optical manipulation with a temporal resolution of a few milliseconds.
DOI
For a precise manipulation of particles and cells with laser light as well as for the understanding and the control of the underlying processes it is important to visualize and quantify the response of the specimens. Thus, we investigated if digital holographic microscopy (DHM) can be used in combination with microfluidics to observe optically trapped living cells in a minimally invasive fashion during laser micromanipulation. The obtained results demonstrate that DHM multi-focus phase contrast provides label-free quantitative monitoring of optical manipulation with a temporal resolution of a few milliseconds.
DOI
Insights into the mechanisms of myosin and kinesin molecular motors from the single-molecule unbinding force measurements
Sergey V. Mikhailenko, Yusuke Oguchi and Shin'ichi Ishiwata
In cells, ATP (adenosine triphosphate)-driven motor proteins, both cytoskeletal and nucleic acid-based, operate on their corresponding ‘tracks’, that is, actin, microtubules or nucleic acids, by converting the chemical energy of ATP hydrolysis into mechanical work. During each mechanochemical cycle, a motor proceeds via several nucleotide states, characterized by different affinities for the ‘track’ filament and different nucleotide (ATP or ADP) binding kinetics, which is crucial for a motor to efficiently perform its cellular functions. The measurements of the rupture force between the motor and the track by applying external loads to the individual motor–substrate bonds in various nucleotide states have proved to be an important tool to obtain valuable insights into the mechanism of the motors' performance. We review the application of this technique to various linear molecular motors, both processive and non-processive, giving special attention to the importance of the experimental geometry.
In cells, ATP (adenosine triphosphate)-driven motor proteins, both cytoskeletal and nucleic acid-based, operate on their corresponding ‘tracks’, that is, actin, microtubules or nucleic acids, by converting the chemical energy of ATP hydrolysis into mechanical work. During each mechanochemical cycle, a motor proceeds via several nucleotide states, characterized by different affinities for the ‘track’ filament and different nucleotide (ATP or ADP) binding kinetics, which is crucial for a motor to efficiently perform its cellular functions. The measurements of the rupture force between the motor and the track by applying external loads to the individual motor–substrate bonds in various nucleotide states have proved to be an important tool to obtain valuable insights into the mechanism of the motors' performance. We review the application of this technique to various linear molecular motors, both processive and non-processive, giving special attention to the importance of the experimental geometry.
Two-dimensional dielectrophoretic particle trapping in a hybrid crystal/PDMS-system
Michael Esseling, Frank Holtmann, Mike Woerdemann, and Cornelia Denz
Dielectrophoretic forces originating from highly modulated electric fields can be used to trap particles on surfaces. An all-optical way to induce such fields is the use of a photorefractive material, where the fields that modulate the refractive index are present at the surface. We present a method for two-dimensional particle alignment on an optically structured photorefractive lithium niobate crystal. The structuring is done using an amplitude-modulating spatial light modulator and laser illumination. We demonstrate trapping of uncharged graphite particles in periodic and arbitrary patterns and provide a discussion of the limitations and the necessary boundary conditions for maximum trapping efficiency. The photorefractive crystal is utilized as bottom part of a PDMS channel in order to demonstrate two-dimensional dielectrophoretic trapping in a microfluidic system.
DOI
Dielectrophoretic forces originating from highly modulated electric fields can be used to trap particles on surfaces. An all-optical way to induce such fields is the use of a photorefractive material, where the fields that modulate the refractive index are present at the surface. We present a method for two-dimensional particle alignment on an optically structured photorefractive lithium niobate crystal. The structuring is done using an amplitude-modulating spatial light modulator and laser illumination. We demonstrate trapping of uncharged graphite particles in periodic and arbitrary patterns and provide a discussion of the limitations and the necessary boundary conditions for maximum trapping efficiency. The photorefractive crystal is utilized as bottom part of a PDMS channel in order to demonstrate two-dimensional dielectrophoretic trapping in a microfluidic system.
DOI
Single cell optical transfection
David J. Stevenson, Frank J. Gunn-Moore, Paul Campbell and Kishan Dholakia
The plasma membrane of a eukaryotic cell is impermeable to most hydrophilic substances, yet the insertion of these materials into cells is an extremely important and universal requirement for the cell biologist. To address this need, many transfection techniques have been developed including viral, lipoplex, polyplex, capillary microinjection, gene gun and electroporation. The current discussion explores a procedure called optical injection, where a laser field transiently increases the membrane permeability to allow species to be internalized. If the internalized substance is a nucleic acid, such as DNA, RNA or small interfering RNA (siRNA), then the process is called optical transfection. This contactless, aseptic, single cell transfection method provides a key nanosurgical tool to the microscopist—the intracellular delivery of reagents and single nanoscopic objects. The experimental possibilities enabled by this technology are only beginning to be realized. A review of optical transfection is presented, along with a forecast of future applications of this rapidly developing and exciting technology.
The plasma membrane of a eukaryotic cell is impermeable to most hydrophilic substances, yet the insertion of these materials into cells is an extremely important and universal requirement for the cell biologist. To address this need, many transfection techniques have been developed including viral, lipoplex, polyplex, capillary microinjection, gene gun and electroporation. The current discussion explores a procedure called optical injection, where a laser field transiently increases the membrane permeability to allow species to be internalized. If the internalized substance is a nucleic acid, such as DNA, RNA or small interfering RNA (siRNA), then the process is called optical transfection. This contactless, aseptic, single cell transfection method provides a key nanosurgical tool to the microscopist—the intracellular delivery of reagents and single nanoscopic objects. The experimental possibilities enabled by this technology are only beginning to be realized. A review of optical transfection is presented, along with a forecast of future applications of this rapidly developing and exciting technology.
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