In vivo three-photon microscopy of subcortical structures within an intact mouse brain
Two-photon fluorescence microscopy 1 enables scientists in various fields including neuroscience 2 , 3 , embryology 4 and oncology 5 to visualize in vivo and ex vivo tissue morphology and physiology at a cellular level deep within scattering tissue. However, tissue scattering limits the maximum imag...
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| Veröffentlicht in: | Nature photonics 2013-03, Vol.7 (3), p.205-209 |
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| creator | Horton, Nicholas G. Wang, Ke Kobat, Demirhan Clark, Catharine G. Wise, Frank W. Schaffer, Chris B. Xu, Chris |
| description | Two-photon fluorescence microscopy
1
enables scientists in various fields including neuroscience
2
,
3
, embryology
4
and oncology
5
to visualize
in vivo
and
ex vivo
tissue morphology and physiology at a cellular level deep within scattering tissue. However, tissue scattering limits the maximum imaging depth of two-photon fluorescence microscopy to the cortical layer within mouse brain, and imaging subcortical structures currently requires the removal of overlying brain tissue
3
or the insertion of optical probes
6
,
7
. Here, we demonstrate non-invasive, high-resolution,
in vivo
imaging of subcortical structures within an intact mouse brain using three-photon fluorescence microscopy at a spectral excitation window of 1,700 nm. Vascular structures as well as red fluorescent protein-labelled neurons within the mouse hippocampus are imaged. The combination of the long excitation wavelength and the higher-order nonlinear excitation overcomes the limitations of two-photon fluorescence microscopy, enabling biological investigations to take place at a greater depth within tissue.
Three-photon microscopy performed at the infrared wavelength of 1,700 nm makes it possible to image hard-to-reach vascular structures and labelled neurons in the hippocampus of a mouse brain. |
| doi_str_mv | 10.1038/nphoton.2012.336 |
| format | Article |
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1
enables scientists in various fields including neuroscience
2
,
3
, embryology
4
and oncology
5
to visualize
in vivo
and
ex vivo
tissue morphology and physiology at a cellular level deep within scattering tissue. However, tissue scattering limits the maximum imaging depth of two-photon fluorescence microscopy to the cortical layer within mouse brain, and imaging subcortical structures currently requires the removal of overlying brain tissue
3
or the insertion of optical probes
6
,
7
. Here, we demonstrate non-invasive, high-resolution,
in vivo
imaging of subcortical structures within an intact mouse brain using three-photon fluorescence microscopy at a spectral excitation window of 1,700 nm. Vascular structures as well as red fluorescent protein-labelled neurons within the mouse hippocampus are imaged. The combination of the long excitation wavelength and the higher-order nonlinear excitation overcomes the limitations of two-photon fluorescence microscopy, enabling biological investigations to take place at a greater depth within tissue.
Three-photon microscopy performed at the infrared wavelength of 1,700 nm makes it possible to image hard-to-reach vascular structures and labelled neurons in the hippocampus of a mouse brain.</description><identifier>ISSN: 1749-4885</identifier><identifier>EISSN: 1749-4893</identifier><identifier>DOI: 10.1038/nphoton.2012.336</identifier><identifier>PMID: 24353743</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>639/624/1107/328/2057 ; 639/624/1111/55 ; Applied and Technical Physics ; Embryology ; Fluorescence ; Fluorescence microscopy ; letter ; Microscopy ; Photonics ; Physics ; Quantum Physics ; Tissues</subject><ispartof>Nature photonics, 2013-03, Vol.7 (3), p.205-209</ispartof><rights>Springer Nature Limited 2013</rights><rights>Copyright Nature Publishing Group Mar 2013</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c612t-2b2558ebf09ce3570b5f6e7dfdbd22248cf0e62809fa9ef9c0e97e4430fcad083</citedby><cites>FETCH-LOGICAL-c612t-2b2558ebf09ce3570b5f6e7dfdbd22248cf0e62809fa9ef9c0e97e4430fcad083</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nphoton.2012.336$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nphoton.2012.336$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>230,314,776,780,881,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24353743$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Horton, Nicholas G.</creatorcontrib><creatorcontrib>Wang, Ke</creatorcontrib><creatorcontrib>Kobat, Demirhan</creatorcontrib><creatorcontrib>Clark, Catharine G.</creatorcontrib><creatorcontrib>Wise, Frank W.</creatorcontrib><creatorcontrib>Schaffer, Chris B.</creatorcontrib><creatorcontrib>Xu, Chris</creatorcontrib><title>In vivo three-photon microscopy of subcortical structures within an intact mouse brain</title><title>Nature photonics</title><addtitle>Nature Photon</addtitle><addtitle>Nat Photonics</addtitle><description>Two-photon fluorescence microscopy
1
enables scientists in various fields including neuroscience
2
,
3
, embryology
4
and oncology
5
to visualize
in vivo
and
ex vivo
tissue morphology and physiology at a cellular level deep within scattering tissue. However, tissue scattering limits the maximum imaging depth of two-photon fluorescence microscopy to the cortical layer within mouse brain, and imaging subcortical structures currently requires the removal of overlying brain tissue
3
or the insertion of optical probes
6
,
7
. Here, we demonstrate non-invasive, high-resolution,
in vivo
imaging of subcortical structures within an intact mouse brain using three-photon fluorescence microscopy at a spectral excitation window of 1,700 nm. Vascular structures as well as red fluorescent protein-labelled neurons within the mouse hippocampus are imaged. The combination of the long excitation wavelength and the higher-order nonlinear excitation overcomes the limitations of two-photon fluorescence microscopy, enabling biological investigations to take place at a greater depth within tissue.
Three-photon microscopy performed at the infrared wavelength of 1,700 nm makes it possible to image hard-to-reach vascular structures and labelled neurons in the hippocampus of a mouse brain.</description><subject>639/624/1107/328/2057</subject><subject>639/624/1111/55</subject><subject>Applied and Technical Physics</subject><subject>Embryology</subject><subject>Fluorescence</subject><subject>Fluorescence microscopy</subject><subject>letter</subject><subject>Microscopy</subject><subject>Photonics</subject><subject>Physics</subject><subject>Quantum Physics</subject><subject>Tissues</subject><issn>1749-4885</issn><issn>1749-4893</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>BENPR</sourceid><recordid>eNqFkc9rFDEUx4MotlbvniTgxcus-TUzyUWQorZQ8KJeQybz0k2ZSdYks9L_vhl2XaognhJ4n_dJ3vsi9JqSDSVcvg-7bSwxbBihbMN59wSd016oRkjFn57usj1DL3K-I6TlirHn6IwJ3vJe8HP04zrgvd9HXLYJoDn48OxtitnG3T2ODudlsDEVb82Ec0mLLUuCjH_5svUBm4B9KMYWPMclAx6S8eEleubMlOHV8bxA3z9_-nZ51dx8_XJ9-fGmsR1lpWEDa1sJgyPKAm97MrSug3504zAyxoS0jkDHJFHOKHDKElA9CMGJs2Ykkl-gDwfvbhlmGC2Eksykd8nPJt3raLz-sxL8Vt_GveayE7JnVfDuKEjx5wK56NlnC9NkAtRxNJWsa-umKP0_yhlnggm1Wt_-hd7FJYW6iUqtnFRiFZIDtS47J3Cnf1Oi13z1MV-95qtrvrXlzeN5Tw2_A60APQC5lsItpEcv_0v6ANdhtZY</recordid><startdate>20130301</startdate><enddate>20130301</enddate><creator>Horton, Nicholas G.</creator><creator>Wang, Ke</creator><creator>Kobat, Demirhan</creator><creator>Clark, Catharine G.</creator><creator>Wise, Frank W.</creator><creator>Schaffer, Chris B.</creator><creator>Xu, Chris</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>7SP</scope><scope>7U5</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>GNUQQ</scope><scope>H8D</scope><scope>HCIFZ</scope><scope>L7M</scope><scope>LK8</scope><scope>M7P</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PHGZM</scope><scope>PHGZT</scope><scope>PKEHL</scope><scope>PQEST</scope><scope>PQGLB</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>7TK</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20130301</creationdate><title>In vivo three-photon microscopy of subcortical structures within an intact mouse brain</title><author>Horton, Nicholas G. ; 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1
enables scientists in various fields including neuroscience
2
,
3
, embryology
4
and oncology
5
to visualize
in vivo
and
ex vivo
tissue morphology and physiology at a cellular level deep within scattering tissue. However, tissue scattering limits the maximum imaging depth of two-photon fluorescence microscopy to the cortical layer within mouse brain, and imaging subcortical structures currently requires the removal of overlying brain tissue
3
or the insertion of optical probes
6
,
7
. Here, we demonstrate non-invasive, high-resolution,
in vivo
imaging of subcortical structures within an intact mouse brain using three-photon fluorescence microscopy at a spectral excitation window of 1,700 nm. Vascular structures as well as red fluorescent protein-labelled neurons within the mouse hippocampus are imaged. The combination of the long excitation wavelength and the higher-order nonlinear excitation overcomes the limitations of two-photon fluorescence microscopy, enabling biological investigations to take place at a greater depth within tissue.
Three-photon microscopy performed at the infrared wavelength of 1,700 nm makes it possible to image hard-to-reach vascular structures and labelled neurons in the hippocampus of a mouse brain.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>24353743</pmid><doi>10.1038/nphoton.2012.336</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1749-4885 |
| ispartof | Nature photonics, 2013-03, Vol.7 (3), p.205-209 |
| issn | 1749-4885 1749-4893 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3864872 |
| source | Springer Nature - Complete Springer Journals; Nature Journals Online |
| subjects | 639/624/1107/328/2057 639/624/1111/55 Applied and Technical Physics Embryology Fluorescence Fluorescence microscopy letter Microscopy Photonics Physics Quantum Physics Tissues |
| title | In vivo three-photon microscopy of subcortical structures within an intact mouse brain |
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