Eintrag weiter verarbeiten
Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V
Gespeichert in:
| Zeitschriftentitel: | Proceedings of the National Academy of Sciences |
|---|---|
| Personen und Körperschaften: | , , , , , |
| In: | Proceedings of the National Academy of Sciences, 109, 2012, 14, S. 5294-5298 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Proceedings of the National Academy of Sciences
|
| Schlagwörter: |
| author_facet |
Ohmachi, Masashi Komori, Yasunori Iwane, Atsuko H. Fujii, Fumihiko Jin, Takashi Yanagida, Toshio Ohmachi, Masashi Komori, Yasunori Iwane, Atsuko H. Fujii, Fumihiko Jin, Takashi Yanagida, Toshio |
|---|---|
| author |
Ohmachi, Masashi Komori, Yasunori Iwane, Atsuko H. Fujii, Fumihiko Jin, Takashi Yanagida, Toshio |
| spellingShingle |
Ohmachi, Masashi Komori, Yasunori Iwane, Atsuko H. Fujii, Fumihiko Jin, Takashi Yanagida, Toshio Proceedings of the National Academy of Sciences Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V Multidisciplinary |
| author_sort |
ohmachi, masashi |
| spelling |
Ohmachi, Masashi Komori, Yasunori Iwane, Atsuko H. Fujii, Fumihiko Jin, Takashi Yanagida, Toshio 0027-8424 1091-6490 Proceedings of the National Academy of Sciences Multidisciplinary http://dx.doi.org/10.1073/pnas.1118472109 <jats:p>Single molecule fluorescence polarization techniques have been used for three-dimensional (3D) orientation measurements to observe the dynamic properties of single molecules. However, only few techniques can simultaneously measure 3D orientation and position. Furthermore, these techniques often require complex equipment and cumbersome analysis. We have developed a microscopy system and synthesized highly fluorescent, rod-like shaped quantum dots (Q rods), which have linear polarizations, to simultaneously measure the position and 3D orientation of a single fluorescent probe. The optics splits the fluorescence from the probe into four different spots depending on the polarization angle and projects them onto a CCD camera. These spots are used to determine the 2D position and 3D orientation. Q rod orientations could be determined with better than 10° accuracy at 33 ms time resolution. We applied our microscopy and Q rods to simultaneously measure myosin V movement along an actin filament and rotation around its own axis, finding that myosin V rotates 90° for each step. From this result, we suggest that in the two-headed bound state, myosin V necks are perpendicular to one another, while in the one-headed bound state the detached trailing myosin V head is biased forward in part by rotating its lever arm about its own axis. This microscopy system should be applicable to a wide range of dynamic biological processes that depend on single molecule orientation dynamics.</jats:p> Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V Proceedings of the National Academy of Sciences |
| doi_str_mv |
10.1073/pnas.1118472109 |
| facet_avail |
Online Free |
| format |
ElectronicArticle |
| fullrecord |
blob:ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTA3My9wbmFzLjExMTg0NzIxMDk |
| id |
ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTA3My9wbmFzLjExMTg0NzIxMDk |
| institution |
DE-Zi4 DE-Gla1 DE-15 DE-Pl11 DE-Rs1 DE-14 DE-105 DE-Ch1 DE-L229 DE-D275 DE-Bn3 DE-Brt1 DE-Zwi2 DE-D161 |
| imprint |
Proceedings of the National Academy of Sciences, 2012 |
| imprint_str_mv |
Proceedings of the National Academy of Sciences, 2012 |
| issn |
0027-8424 1091-6490 |
| issn_str_mv |
0027-8424 1091-6490 |
| language |
English |
| mega_collection |
Proceedings of the National Academy of Sciences (CrossRef) |
| match_str |
ohmachi2012fluorescencemicroscopyforsimultaneousobservationof3dorientationandmovementanditsapplicationtoquantumrodtaggedmyosinv |
| publishDateSort |
2012 |
| publisher |
Proceedings of the National Academy of Sciences |
| recordtype |
ai |
| record_format |
ai |
| series |
Proceedings of the National Academy of Sciences |
| source_id |
49 |
| title |
Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V |
| title_unstemmed |
Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V |
| title_full |
Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V |
| title_fullStr |
Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V |
| title_full_unstemmed |
Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V |
| title_short |
Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V |
| title_sort |
fluorescence microscopy for simultaneous observation of 3d orientation and movement and its application to quantum rod-tagged myosin v |
| topic |
Multidisciplinary |
| url |
http://dx.doi.org/10.1073/pnas.1118472109 |
| publishDate |
2012 |
| physical |
5294-5298 |
| description |
<jats:p>Single molecule fluorescence polarization techniques have been used for three-dimensional (3D) orientation measurements to observe the dynamic properties of single molecules. However, only few techniques can simultaneously measure 3D orientation and position. Furthermore, these techniques often require complex equipment and cumbersome analysis. We have developed a microscopy system and synthesized highly fluorescent, rod-like shaped quantum dots (Q rods), which have linear polarizations, to simultaneously measure the position and 3D orientation of a single fluorescent probe. The optics splits the fluorescence from the probe into four different spots depending on the polarization angle and projects them onto a CCD camera. These spots are used to determine the 2D position and 3D orientation. Q rod orientations could be determined with better than 10° accuracy at 33 ms time resolution. We applied our microscopy and Q rods to simultaneously measure myosin V movement along an actin filament and rotation around its own axis, finding that myosin V rotates 90° for each step. From this result, we suggest that in the two-headed bound state, myosin V necks are perpendicular to one another, while in the one-headed bound state the detached trailing myosin V head is biased forward in part by rotating its lever arm about its own axis. This microscopy system should be applicable to a wide range of dynamic biological processes that depend on single molecule orientation dynamics.</jats:p> |
| container_issue |
14 |
| container_start_page |
5294 |
| container_title |
Proceedings of the National Academy of Sciences |
| container_volume |
109 |
| format_de105 |
Article, E-Article |
| format_de14 |
Article, E-Article |
| format_de15 |
Article, E-Article |
| format_de520 |
Article, E-Article |
| format_de540 |
Article, E-Article |
| format_dech1 |
Article, E-Article |
| format_ded117 |
Article, E-Article |
| format_degla1 |
E-Article |
| format_del152 |
Buch |
| format_del189 |
Article, E-Article |
| format_dezi4 |
Article |
| format_dezwi2 |
Article, E-Article |
| format_finc |
Article, E-Article |
| format_nrw |
Article, E-Article |
| _version_ |
1792337728113737728 |
| geogr_code |
not assigned |
| last_indexed |
2024-03-01T15:20:57.47Z |
| geogr_code_person |
not assigned |
| openURL |
url_ver=Z39.88-2004&ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fvufind.svn.sourceforge.net%3Agenerator&rft.title=Fluorescence+microscopy+for+simultaneous+observation+of+3D+orientation+and+movement+and+its+application+to+quantum+rod-tagged+myosin+V&rft.date=2012-04-03&genre=article&issn=1091-6490&volume=109&issue=14&spage=5294&epage=5298&pages=5294-5298&jtitle=Proceedings+of+the+National+Academy+of+Sciences&atitle=Fluorescence+microscopy+for+simultaneous+observation+of+3D+orientation+and+movement+and+its+application+to+quantum+rod-tagged+myosin+V&aulast=Yanagida&aufirst=Toshio&rft_id=info%3Adoi%2F10.1073%2Fpnas.1118472109&rft.language%5B0%5D=eng |
| SOLR | |
| _version_ | 1792337728113737728 |
| author | Ohmachi, Masashi, Komori, Yasunori, Iwane, Atsuko H., Fujii, Fumihiko, Jin, Takashi, Yanagida, Toshio |
| author_facet | Ohmachi, Masashi, Komori, Yasunori, Iwane, Atsuko H., Fujii, Fumihiko, Jin, Takashi, Yanagida, Toshio, Ohmachi, Masashi, Komori, Yasunori, Iwane, Atsuko H., Fujii, Fumihiko, Jin, Takashi, Yanagida, Toshio |
| author_sort | ohmachi, masashi |
| container_issue | 14 |
| container_start_page | 5294 |
| container_title | Proceedings of the National Academy of Sciences |
| container_volume | 109 |
| description | <jats:p>Single molecule fluorescence polarization techniques have been used for three-dimensional (3D) orientation measurements to observe the dynamic properties of single molecules. However, only few techniques can simultaneously measure 3D orientation and position. Furthermore, these techniques often require complex equipment and cumbersome analysis. We have developed a microscopy system and synthesized highly fluorescent, rod-like shaped quantum dots (Q rods), which have linear polarizations, to simultaneously measure the position and 3D orientation of a single fluorescent probe. The optics splits the fluorescence from the probe into four different spots depending on the polarization angle and projects them onto a CCD camera. These spots are used to determine the 2D position and 3D orientation. Q rod orientations could be determined with better than 10° accuracy at 33 ms time resolution. We applied our microscopy and Q rods to simultaneously measure myosin V movement along an actin filament and rotation around its own axis, finding that myosin V rotates 90° for each step. From this result, we suggest that in the two-headed bound state, myosin V necks are perpendicular to one another, while in the one-headed bound state the detached trailing myosin V head is biased forward in part by rotating its lever arm about its own axis. This microscopy system should be applicable to a wide range of dynamic biological processes that depend on single molecule orientation dynamics.</jats:p> |
| doi_str_mv | 10.1073/pnas.1118472109 |
| facet_avail | Online, Free |
| format | ElectronicArticle |
| format_de105 | Article, E-Article |
| format_de14 | Article, E-Article |
| format_de15 | Article, E-Article |
| format_de520 | Article, E-Article |
| format_de540 | Article, E-Article |
| format_dech1 | Article, E-Article |
| format_ded117 | Article, E-Article |
| format_degla1 | E-Article |
| format_del152 | Buch |
| format_del189 | Article, E-Article |
| format_dezi4 | Article |
| format_dezwi2 | Article, E-Article |
| format_finc | Article, E-Article |
| format_nrw | Article, E-Article |
| geogr_code | not assigned |
| geogr_code_person | not assigned |
| id | ai-49-aHR0cDovL2R4LmRvaS5vcmcvMTAuMTA3My9wbmFzLjExMTg0NzIxMDk |
| imprint | Proceedings of the National Academy of Sciences, 2012 |
| imprint_str_mv | Proceedings of the National Academy of Sciences, 2012 |
| institution | DE-Zi4, DE-Gla1, DE-15, DE-Pl11, DE-Rs1, DE-14, DE-105, DE-Ch1, DE-L229, DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161 |
| issn | 0027-8424, 1091-6490 |
| issn_str_mv | 0027-8424, 1091-6490 |
| language | English |
| last_indexed | 2024-03-01T15:20:57.47Z |
| match_str | ohmachi2012fluorescencemicroscopyforsimultaneousobservationof3dorientationandmovementanditsapplicationtoquantumrodtaggedmyosinv |
| mega_collection | Proceedings of the National Academy of Sciences (CrossRef) |
| physical | 5294-5298 |
| publishDate | 2012 |
| publishDateSort | 2012 |
| publisher | Proceedings of the National Academy of Sciences |
| record_format | ai |
| recordtype | ai |
| series | Proceedings of the National Academy of Sciences |
| source_id | 49 |
| spelling | Ohmachi, Masashi Komori, Yasunori Iwane, Atsuko H. Fujii, Fumihiko Jin, Takashi Yanagida, Toshio 0027-8424 1091-6490 Proceedings of the National Academy of Sciences Multidisciplinary http://dx.doi.org/10.1073/pnas.1118472109 <jats:p>Single molecule fluorescence polarization techniques have been used for three-dimensional (3D) orientation measurements to observe the dynamic properties of single molecules. However, only few techniques can simultaneously measure 3D orientation and position. Furthermore, these techniques often require complex equipment and cumbersome analysis. We have developed a microscopy system and synthesized highly fluorescent, rod-like shaped quantum dots (Q rods), which have linear polarizations, to simultaneously measure the position and 3D orientation of a single fluorescent probe. The optics splits the fluorescence from the probe into four different spots depending on the polarization angle and projects them onto a CCD camera. These spots are used to determine the 2D position and 3D orientation. Q rod orientations could be determined with better than 10° accuracy at 33 ms time resolution. We applied our microscopy and Q rods to simultaneously measure myosin V movement along an actin filament and rotation around its own axis, finding that myosin V rotates 90° for each step. From this result, we suggest that in the two-headed bound state, myosin V necks are perpendicular to one another, while in the one-headed bound state the detached trailing myosin V head is biased forward in part by rotating its lever arm about its own axis. This microscopy system should be applicable to a wide range of dynamic biological processes that depend on single molecule orientation dynamics.</jats:p> Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V Proceedings of the National Academy of Sciences |
| spellingShingle | Ohmachi, Masashi, Komori, Yasunori, Iwane, Atsuko H., Fujii, Fumihiko, Jin, Takashi, Yanagida, Toshio, Proceedings of the National Academy of Sciences, Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V, Multidisciplinary |
| title | Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V |
| title_full | Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V |
| title_fullStr | Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V |
| title_full_unstemmed | Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V |
| title_short | Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V |
| title_sort | fluorescence microscopy for simultaneous observation of 3d orientation and movement and its application to quantum rod-tagged myosin v |
| title_unstemmed | Fluorescence microscopy for simultaneous observation of 3D orientation and movement and its application to quantum rod-tagged myosin V |
| topic | Multidisciplinary |
| url | http://dx.doi.org/10.1073/pnas.1118472109 |