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Literature DB >> 23931522

Circumventing photodamage in live-cell microscopy.

Abstract

Fluorescence microscopy has become an essential tool in cell biology. This technique allows researchers to visualize the dynamics of tissue, cells, individual organelles, and macromolecular assemblies inside the cell. Unfortunately, fluorescence microscopy is not completely "noninvasive" as the high-intensity excitation light required for excitation of fluorophores is inherently toxic for live cells. Physiological changes induced by excessive illumination can lead to artifacts and abnormal responses. In this chapter, we review major factors that contribute to phototoxicity and discuss practical solutions for circumventing photodamage. These solutions include the proper choice of image acquisition parameters, optimization of filter sets, hardware synchronization, and the use of intelligent illumination to avoid unnecessary light exposure.

Entities: CellLine Chemical Disease Gene Species

Keywords: Fluorescence; GFP; Live-cell microscopy; Photodamage; Phototoxicity

Mesh：

Substances：

Year: 2013 PMID： 23931522 PMCID： PMC3843244 DOI： 10.1016/B978-0-12-407761-4.00023-3

Source DB: PubMed Journal: Methods Cell Biol ISSN： 0091-679X Impact factor: 1.441

22 in total

1. Optical sectioning deep inside live embryos by selective plane illumination microscopy.

Authors: Jan Huisken; Jim Swoger; Filippo Del Bene; Joachim Wittbrodt; Ernst H K Stelzer
Journal: Science Date: 2004-08-13 Impact factor: 47.728

2. Speckle microscopic evaluation of microtubule transport in growing nerve processes.

Authors: S Chang; T M Svitkina; G G Borisy; S V Popov
Journal: Nat Cell Biol Date: 1999-11 Impact factor: 28.824

Review 3. The p38-mediated stress-activated checkpoint. A rapid response system for delaying progression through antephase and entry into mitosis.

Authors: Alexei Mikhailov; Mio Shinohara; Conly L Rieder
Journal: Cell Cycle Date: 2005-01-11 Impact factor: 4.534

4. Major signal increase in fluorescence microscopy through dark-state relaxation.

Authors: Gerald Donnert; Christian Eggeling; Stefan W Hell
Journal: Nat Methods Date: 2006-12-10 Impact factor: 28.547

5. Microtubule treadmilling in vivo.

Authors: V I Rodionov; G G Borisy
Journal: Science Date: 1997-01-10 Impact factor: 47.728

6. The sudden recruitment of gamma-tubulin to the centrosome at the onset of mitosis and its dynamic exchange throughout the cell cycle, do not require microtubules.

Authors: A Khodjakov; C L Rieder
Journal: J Cell Biol Date: 1999-08-09 Impact factor: 10.539

Circumventing photodamage in live-cell microscopy.

1. Optical sectioning deep inside live embryos by selective plane illumination microscopy.

2. Speckle microscopic evaluation of microtubule transport in growing nerve processes.

Review 3. The p38-mediated stress-activated checkpoint. A rapid response system for delaying progression through antephase and entry into mitosis.

4. Major signal increase in fluorescence microscopy through dark-state relaxation.

5. Microtubule treadmilling in vivo.

6. The sudden recruitment of gamma-tubulin to the centrosome at the onset of mitosis and its dynamic exchange throughout the cell cycle, do not require microtubules.

7. Centrosomes enhance the fidelity of cytokinesis in vertebrates and are required for cell cycle progression.

8. Kinetochore microtubule dynamics and the metaphase-anaphase transition.

9. A mechanism for nuclear positioning in fission yeast based on microtubule pushing.

10. Noninvasive imaging beyond the diffraction limit of 3D dynamics in thickly fluorescent specimens.

1. Improved two-photon imaging of living neurons in brain tissue through temporal gating.

Review 2. Inside single cells: quantitative analysis with advanced optics and nanomaterials.

3. Expanded palette of Nano-lanterns for real-time multicolor luminescence imaging.

4. Long-term Brillouin imaging of live cells with reduced absorption-mediated damage at 660 nm wavelength.

5. Quantitative assessment of neural outgrowth using spatial light interference microscopy.

6. Super-resolution microscopy for biological specimens: lensless phase retrieval in noisy conditions.

Review 7. Genetically Encoded Voltage Indicators: Opportunities and Challenges.

Review 8. Challenges in long-term imaging and quantification of single-cell dynamics.

Review 9. Spatially Resolved Analytical Chemistry in Intact, Living Tissues.

10. Measuring Synaptic Vesicle Endocytosis in Cultured Hippocampal Neurons.