Laser-mediated dissection methods have been used for many years to micro-irradiate biological samples, but recent technological progress has rendered this technique more precise, powerful, and easy to use. Today pulsed lasers can be operated with diffraction limited, sub-micrometer precision to ablate intracellular structures. Here, we discuss laser nanosurgery setups and the instrumentation in our laboratory. We describe how to use this technique to ablate cytoskeletal elements in living cells. We also show how this technique can be used in multicellular organisms, to micropuncture and/or ablate cells of interest and finally how to monitor a successful laser nanosurgery.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/978-1-4939-6810-7_8 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi.
Microsyst Nanoeng
April 2024
Institute of Physics Belgrade, University of Belgrade, National Institute of the Republic of Serbia, Pregrevica 118, 11080 Belgrade, Serbia.
Studying the membrane physiology of filamentous fungi is key to understanding their interactions with the environment and crucial for developing new therapeutic strategies for disease-causing pathogens. However, their plasma membrane has been inaccessible for a micron-sized patch-clamp pipette for pA current recordings due to the rigid chitinous cell wall. Here, we report the first femtosecond IR laser nanosurgery of the cell wall of the filamentous fungi, which enabled patch-clamp measurements on protoplasts released from hyphae.
View Article and Find Full Text PDFEpithelial restitution in 3D - Revealing biomechanical and physiochemical dynamics in intestinal organoids via fs laser nanosurgery.
iScience
November 2023
Institute of Quantum Optics, Leibniz University Hannover, 30167 Hannover, Germany.
Intestinal organoids represent a three-dimensional cell culture system mimicking the mammalian intestine. The application of single-cell ablation for defined wounding via a femtosecond laser system within the crypt base allowed us to study cell dynamics during epithelial restitution. Neighboring cells formed a contractile actin ring encircling the damaged cell, changed the cellular aspect ratio, and immediately closed the barrier.
View Article and Find Full Text PDFMimicking acute airway tissue damage using femtosecond laser nanosurgery in airway organoids.
Front Cell Dev Biol
September 2023
Institute of Quantum Optics, Leibniz University Hannover, Hannover, Germany.
Airway organoids derived from adult murine epithelial cells represent a complex 3D system mimicking the airway epithelial tissue's native cell composition and physiological properties. In combination with a precise damage induction via femtosecond laser-based nanosurgery, this model might allow for the examination of intra- and intercellular dynamics in the course of repair processes with a high spatio-temporal resolution, which can hardly be reached using approaches. For characterization of the organoids' response to single or multiple-cell ablation, we first analyzed overall organoid survival and found that airway organoids were capable of efficiently repairing damage induced by femtosecond laser-based ablation of a single to ten cells within 24 h.
View Article and Find Full Text PDFA single cell death is disruptive to spontaneous Ca activity in astrocytes.
Front Cell Neurosci
July 2022
Department of Bioengineering, Institute of Engineering in Medicine, University of California, San Diego, San Diego, CA, United States.
Astrocytes in the brain are rapidly recruited to sites of injury where they phagocytose damaged material and take up neurotransmitters and ions to avoid the spreading of damaging molecules. In this study we investigate the calcium (Ca) response in astrocytes to nearby cell death. To induce cell death in a nearby cell we utilized a laser nanosurgery system to photolyze a selected cell from an established astrocyte cell line (Ast1).
View Article and Find Full Text PDFInvestigation of Colonic Regeneration via Precise Damage Application Using Femtosecond Laser-Based Nanosurgery.
Cells
March 2022
Institute of Quantum Optics, Leibniz University Hannover, 30167 Hannover, Germany.
Organoids represent the cellular composition of natural tissue. So called colonoids, organoids derived from colon tissue, are a good model for understanding regeneration. However, next to the cellular composition, the surrounding matrix, the cell-cell interactions, and environmental factors have to be considered.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!