More than double the fun with two-photon excitation microscopy
Abstract For generations researchers have been observing the dynamic processes of life through the lens of a microscope. This has offered tremendous insights into biological phenomena that span multiple orders of time- and length-scales ranging from the pure magic of molecular reorganization at the...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-03-01
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| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-024-06057-0 |
| _version_ | 1797233368486641664 |
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| author | Peter Luu Scott E. Fraser Falk Schneider |
| author_facet | Peter Luu Scott E. Fraser Falk Schneider |
| author_sort | Peter Luu |
| collection | DOAJ |
| description | Abstract For generations researchers have been observing the dynamic processes of life through the lens of a microscope. This has offered tremendous insights into biological phenomena that span multiple orders of time- and length-scales ranging from the pure magic of molecular reorganization at the membrane of immune cells, to cell migration and differentiation during development or wound healing. Standard fluorescence microscopy techniques offer glimpses at such processes in vitro, however, when applied in intact systems, they are challenged by reduced signal strengths and signal-to-noise ratios that result from deeper imaging. As a remedy, two-photon excitation (TPE) microscopy takes a special place, because it allows us to investigate processes in vivo, in their natural environment, even in a living animal. Here, we review the fundamental principles underlying TPE aimed at basic and advanced microscopy users interested in adopting TPE for intravital imaging. We focus on applications in neurobiology, present current trends towards faster, wider and deeper imaging, discuss the combination with photon counting technologies for metabolic imaging and spectroscopy, as well as highlight outstanding issues and drawbacks in development and application of these methodologies. |
| first_indexed | 2024-04-24T16:15:04Z |
| format | Article |
| id | doaj.art-0c91252f46e94ffbbbff264b4994bd6f |
| institution | Directory Open Access Journal |
| issn | 2399-3642 |
| language | English |
| last_indexed | 2024-04-24T16:15:04Z |
| publishDate | 2024-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Biology |
| spelling | doaj.art-0c91252f46e94ffbbbff264b4994bd6f2024-03-31T11:29:12ZengNature PortfolioCommunications Biology2399-36422024-03-017111510.1038/s42003-024-06057-0More than double the fun with two-photon excitation microscopyPeter Luu0Scott E. Fraser1Falk Schneider2Translational Imaging Center, Michelson Center for Convergent Bioscience, University of Southern CaliforniaTranslational Imaging Center, Michelson Center for Convergent Bioscience, University of Southern CaliforniaTranslational Imaging Center, Michelson Center for Convergent Bioscience, University of Southern CaliforniaAbstract For generations researchers have been observing the dynamic processes of life through the lens of a microscope. This has offered tremendous insights into biological phenomena that span multiple orders of time- and length-scales ranging from the pure magic of molecular reorganization at the membrane of immune cells, to cell migration and differentiation during development or wound healing. Standard fluorescence microscopy techniques offer glimpses at such processes in vitro, however, when applied in intact systems, they are challenged by reduced signal strengths and signal-to-noise ratios that result from deeper imaging. As a remedy, two-photon excitation (TPE) microscopy takes a special place, because it allows us to investigate processes in vivo, in their natural environment, even in a living animal. Here, we review the fundamental principles underlying TPE aimed at basic and advanced microscopy users interested in adopting TPE for intravital imaging. We focus on applications in neurobiology, present current trends towards faster, wider and deeper imaging, discuss the combination with photon counting technologies for metabolic imaging and spectroscopy, as well as highlight outstanding issues and drawbacks in development and application of these methodologies.https://doi.org/10.1038/s42003-024-06057-0 |
| spellingShingle | Peter Luu Scott E. Fraser Falk Schneider More than double the fun with two-photon excitation microscopy Communications Biology |
| title | More than double the fun with two-photon excitation microscopy |
| title_full | More than double the fun with two-photon excitation microscopy |
| title_fullStr | More than double the fun with two-photon excitation microscopy |
| title_full_unstemmed | More than double the fun with two-photon excitation microscopy |
| title_short | More than double the fun with two-photon excitation microscopy |
| title_sort | more than double the fun with two photon excitation microscopy |
| url | https://doi.org/10.1038/s42003-024-06057-0 |
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