The Benefits of Unnatural Amino Acid Incorporation as Protein Labels for Single Molecule Localization Microscopy
Single Molecule Localization Microscopy (SMLM) is an imaging method that allows for the visualization of structures smaller than the diffraction limit of light (~200 nm). This is achieved through techniques such as stochastic optical reconstruction microscopy (STORM) and photoactivated localization...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2021-03-01
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| Series: | Frontiers in Chemistry |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fchem.2021.641355/full |
| _version_ | 1828896634919976960 |
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| author | Pooja Laxman Shirin Ansari Katharina Gaus Jesse Goyette |
| author_facet | Pooja Laxman Shirin Ansari Katharina Gaus Jesse Goyette |
| author_sort | Pooja Laxman |
| collection | DOAJ |
| description | Single Molecule Localization Microscopy (SMLM) is an imaging method that allows for the visualization of structures smaller than the diffraction limit of light (~200 nm). This is achieved through techniques such as stochastic optical reconstruction microscopy (STORM) and photoactivated localization microscopy (PALM). A large part of obtaining ideal imaging of single molecules is the choice of the right fluorescent label. An upcoming field of protein labeling is incorporating unnatural amino acids (UAAs) with an attached fluorescent dye for precise localization and visualization of individual molecules. For this technique, fluorescent probes are conjugated to UAAs and are introduced into the protein of interest (POI) as a label. Here we contrast this labeling method with other commonly used protein-based labeling methods such as fluorescent proteins (FPs) or self-labeling tags such as Halotag, SNAP-tags, and CLIP-tags, and highlight the benefits and shortcomings of the site-specific incorporation of UAAs coupled with fluorescent dyes in SMLM. |
| first_indexed | 2024-12-13T14:44:20Z |
| format | Article |
| id | doaj.art-0709080b2d2b4d2b999b0a91d1f8b95f |
| institution | Directory Open Access Journal |
| issn | 2296-2646 |
| language | English |
| last_indexed | 2024-12-13T14:44:20Z |
| publishDate | 2021-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Chemistry |
| spelling | doaj.art-0709080b2d2b4d2b999b0a91d1f8b95f2022-12-21T23:41:30ZengFrontiers Media S.A.Frontiers in Chemistry2296-26462021-03-01910.3389/fchem.2021.641355641355The Benefits of Unnatural Amino Acid Incorporation as Protein Labels for Single Molecule Localization MicroscopyPooja LaxmanShirin AnsariKatharina GausJesse GoyetteSingle Molecule Localization Microscopy (SMLM) is an imaging method that allows for the visualization of structures smaller than the diffraction limit of light (~200 nm). This is achieved through techniques such as stochastic optical reconstruction microscopy (STORM) and photoactivated localization microscopy (PALM). A large part of obtaining ideal imaging of single molecules is the choice of the right fluorescent label. An upcoming field of protein labeling is incorporating unnatural amino acids (UAAs) with an attached fluorescent dye for precise localization and visualization of individual molecules. For this technique, fluorescent probes are conjugated to UAAs and are introduced into the protein of interest (POI) as a label. Here we contrast this labeling method with other commonly used protein-based labeling methods such as fluorescent proteins (FPs) or self-labeling tags such as Halotag, SNAP-tags, and CLIP-tags, and highlight the benefits and shortcomings of the site-specific incorporation of UAAs coupled with fluorescent dyes in SMLM.https://www.frontiersin.org/articles/10.3389/fchem.2021.641355/fullunnatural (non-canonical) amino acidssingle molecule localization microscopyfluorescent proteinunnatural amino acid incorporationstochastic optical reconstruction microscopyphoto-activated localization microscopy |
| spellingShingle | Pooja Laxman Shirin Ansari Katharina Gaus Jesse Goyette The Benefits of Unnatural Amino Acid Incorporation as Protein Labels for Single Molecule Localization Microscopy Frontiers in Chemistry unnatural (non-canonical) amino acids single molecule localization microscopy fluorescent protein unnatural amino acid incorporation stochastic optical reconstruction microscopy photo-activated localization microscopy |
| title | The Benefits of Unnatural Amino Acid Incorporation as Protein Labels for Single Molecule Localization Microscopy |
| title_full | The Benefits of Unnatural Amino Acid Incorporation as Protein Labels for Single Molecule Localization Microscopy |
| title_fullStr | The Benefits of Unnatural Amino Acid Incorporation as Protein Labels for Single Molecule Localization Microscopy |
| title_full_unstemmed | The Benefits of Unnatural Amino Acid Incorporation as Protein Labels for Single Molecule Localization Microscopy |
| title_short | The Benefits of Unnatural Amino Acid Incorporation as Protein Labels for Single Molecule Localization Microscopy |
| title_sort | benefits of unnatural amino acid incorporation as protein labels for single molecule localization microscopy |
| topic | unnatural (non-canonical) amino acids single molecule localization microscopy fluorescent protein unnatural amino acid incorporation stochastic optical reconstruction microscopy photo-activated localization microscopy |
| url | https://www.frontiersin.org/articles/10.3389/fchem.2021.641355/full |
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