Directed Positioning of Micronuclei in <i>Paramecium tetraurelia</i> with Laser Tweezers: Absence of Detectible Damage After Manipulation

Aufderheide, Karl J.; Du, Qingguo; Fry, Edward S.

doi:10.1111/j.1550-7408.1993.tb04476.x

Cited by 16 publications

(5 citation statements)

References 19 publications

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“…In addition to qualitative observations, the MMC is useful for quantitative measurements of immobilized specimens as well as various other kinds of microsurgery and manipulations (Aufderheide et al, 1992, 1993). The open nature of the device’s microfluidics system allow the experimenter to position and hold worms, for example, in a custom-designed trapping structure or “bed” microfabricated using PDMS (Fig.…”

Section: Discussionmentioning

confidence: 99%

A Microfluidic-Enabled Mechanical Microcompressor for the Immobilization of Live Single- and Multi-Cellular Specimens

et al. 2014

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A microcompressor is a precision mechanical device that flattens and immobilizes living cells and small organisms for optical microscopy, allowing enhanced visualization of sub-cellular structures and organelles. We have developed an easily fabricated device, which can be equipped with microfluidics, permitting the addition of media or chemicals during observation. This device can be used on both upright and inverted microscopes. The apparatus permits micrometer precision flattening for nondestructive immobilization of specimens as small as a bacterium, while also accommodating larger specimens, such as Caenorhabditis elegans, for long-term observations. The compressor mount is removable and allows easy specimen addition and recovery for later observation. Several customized specimen beds can be incorporated into the base. To demonstrate the capabilities of the device, we have imaged numerous cellular events in several protozoan species, in yeast cells, and in Drosophila melanogaster embryos. We have been able to document previously unreported events, and also perform photobleaching experiments, in conjugating Tetrahymena thermophila.

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Section: Discussionmentioning

confidence: 99%

A Microfluidic-Enabled Mechanical Microcompressor for the Immobilization of Live Single- and Multi-Cellular Specimens

et al. 2014

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“…These forces are sufficient to draw cells or organelles into the focus of the laser beam where they are trapped (for reviews, see: Greulich and Weber 1992;Weber and Greulich 1992;Simmons and Finer 1993). The optical tweezers can be applied to manipulate living cells without detectible damage (Aufderheide et al 1993). We have previously communicated our ability to use optical tweezers to move organelles in living protists and to study the viscoelastic properties of artificial cytoplasmic strands .…”

Section: Introductionmentioning

confidence: 99%

Micromanipulation of statoliths in gravity-sensing Chara rhizoids by optical tweezers

Leitz

Schnepf

Greulich

1995

Planta

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Infrared laser traps (optical tweezers) were used to micromanipulate statoliths in gravity-sensing rhizoids of the green alga Chara vulgaris Vail. We were able to hold and move statoliths with high accuracy and to observe directly the effects of statolith position on cell growth in horizontally positioned rhizoids. The first step in gravitropism, namely the physical action of gravity on statoliths, can be simulated by optical tweezers. The direct laser microirradiation of the rhizoid apex did not cause any visible damage to the cells. Through lateral positioning of statoliths a differential growth of the opposite flank of the cell wall could be induced, corresponding to bending growth in gravitropism. The acropetal displacement of the statolith complex into the extreme apex of the rhizoid caused a temporary decrease in cell growth rate. The rhizoids regained normal growth after remigration of the statoliths to their initial position 10-30 micrometers basal to the rhizoid apex. During basipetal displacement of statoliths, cell growth continued and the statoliths remigrated towards the rhizoid tip after release from the optical trap. The resistance to statolith displacement increased towards the nucleus. The basipetal displacement of the whole complex of statoliths for a long distance (>100 micrometers) caused an increase in cell diameter and a subsequent regaining of normal growth after the statoliths reappeared in the rhizoid apex. We conclude that the statolith displacement interferes with the mechanism of tip growth, i.e. with the transport of Golgi vesicles, either directly by mechanically blocking their flow and/or, indirectly, by disturbing the actomyosin system. In the presence of the actin inhibitor cytochalasin B the optical forces required for acropetal and basipetal displacement of statoliths were significantly reduced to a similar level. The lateral displacement of statoliths was not changed by cytochalasin B. The results indicate: (i) the viscous resistance to optical displacement of statoliths depend mainly on actin, (ii) the lateral displacement of statoliths is not impeded by actin filaments, (iii) the axially directed actin-mediated forces against optical displacement of statoliths (for a distance of 10 micrometers) are stronger in the basipetal than in the acropetal direction, (iv) the forces acting on single statoliths by axially oriented actin filaments are estimated to be in the range of 11-110 pN for acropetal and of 18-180 pN for basipetal statolith displacements.

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“…Then, by moving these crystals until contact was establish with a nucleus, it was possible to push and drag the nucleus around. Moreover, no damage was reported in the organisms during or for some time after interrupting the optical manipulation (Aufderheide et al, 1993). Ketelaar et al (2002) successfully applied the technique directly to nuclei in plant cells when they optically trapped nuclei in Arabidopsis thaliana root hair cells.…”

Section: Genetic Materials Manipulationmentioning

confidence: 99%

Genetic Material Manipulation and Modification by Optical Trapping and Nanosurgery-A Perspective

Blázquez‐Castro

Fernández‐Piqueras

Santos

2020

Front. Bioeng. Biotechnol.

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Light can be employed as a tool to alter and manipulate matter in many ways. An example has been the implementation of optical trapping, the so called optical tweezers, in which light can hold and move small objects with 3D control. Of interest for the Life Sciences and Biotechnology is the fact that biological objects in the size range from tens of nanometers to hundreds of microns can be precisely manipulated through this technology. In particular, it has been shown possible to optically trap and move genetic material (DNA and chromatin) using optical tweezers. Also, these biological entities can be severed, rearranged and reconstructed by the combined use of laser scissors and optical tweezers. In this review, the background, current state and future possibilities of optical tweezers and laser scissors to manipulate, rearrange and alter genetic material (DNA, chromatin and chromosomes) will be presented. Sources of undesirable effects by the optical procedure and measures to avoid them will be discussed. In addition, first tentative approaches at cellular-level genetic and organelle surgery, in which genetic material or DNA-carrying organelles are extracted out or introduced into cells, will be presented.

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Directed Positioning of Micronuclei in Paramecium tetraurelia with Laser Tweezers: Absence of Detectible Damage After Manipulation

Cited by 16 publications

References 19 publications

A Microfluidic-Enabled Mechanical Microcompressor for the Immobilization of Live Single- and Multi-Cellular Specimens

A Microfluidic-Enabled Mechanical Microcompressor for the Immobilization of Live Single- and Multi-Cellular Specimens

Micromanipulation of statoliths in gravity-sensing Chara rhizoids by optical tweezers

Genetic Material Manipulation and Modification by Optical Trapping and Nanosurgery-A Perspective

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