The power of single and multibeam two-photon microscopy for high-resolution and high-speed deep tissue and intravital imaging.

2.50
Hdl Handle:
http://hdl.handle.net/10033/16233
Title:
The power of single and multibeam two-photon microscopy for high-resolution and high-speed deep tissue and intravital imaging.
Authors:
Niesner, Raluca; Andresen, Volker; Neumann, Jens; Spiecker, Heinrich; Gunzer, Matthias
Abstract:
Two-photon microscopy is indispensable for deep tissue and intravital imaging. However, current technology based on single-beam point scanning has reached sensitivity and speed limits because higher performance requires higher laser power leading to sample degradation. We utilize a multifocal scanhead splitting a laser beam into a line of 64 foci, allowing sample illumination in real time at full laser power. This technology requires charge-coupled device field detection in contrast to conventional detection by photomultipliers. A comparison of the optical performance of both setups shows functional equivalence in every measurable parameter down to penetration depths of 200 microm, where most actual experiments are executed. The advantage of photomultiplier detection materializes at imaging depths >300 microm because of their better signal/noise ratio, whereas only charge-coupled devices allow real-time detection of rapid processes (here blood flow). We also find that the point-spread function of both devices strongly depends on tissue constitution and penetration depth. However, employment of a depth-corrected point-spread function allows three-dimensional deconvolution of deep-tissue data up to an image quality resembling surface detection.
Affiliation:
Helmholtz Centre for Infection Research, Junior Research Group Immunodynamics, D-38124 Braunschweig, Germany.
Citation:
The power of single and multibeam two-photon microscopy for high-resolution and high-speed deep tissue and intravital imaging. 2007, 93 (7):2519-29 Biophys. J.
Journal:
Biophysical journal
Issue Date:
1-Oct-2007
URI:
http://hdl.handle.net/10033/16233
DOI:
10.1529/biophysj.106.102459
PubMed ID:
17557785
Type:
Article
Language:
en
ISSN:
0006-3495
Appears in Collections:
Publications of JRG Immunodynamics (ID)

Full metadata record

DC FieldValue Language
dc.contributor.authorNiesner, Raluca-
dc.contributor.authorAndresen, Volker-
dc.contributor.authorNeumann, Jens-
dc.contributor.authorSpiecker, Heinrich-
dc.contributor.authorGunzer, Matthias-
dc.date.accessioned2008-01-17T09:55:02Z-
dc.date.available2008-01-17T09:55:02Z-
dc.date.issued2007-10-01-
dc.identifier.citationThe power of single and multibeam two-photon microscopy for high-resolution and high-speed deep tissue and intravital imaging. 2007, 93 (7):2519-29 Biophys. J.en
dc.identifier.issn0006-3495-
dc.identifier.pmid17557785-
dc.identifier.doi10.1529/biophysj.106.102459-
dc.identifier.urihttp://hdl.handle.net/10033/16233-
dc.description.abstractTwo-photon microscopy is indispensable for deep tissue and intravital imaging. However, current technology based on single-beam point scanning has reached sensitivity and speed limits because higher performance requires higher laser power leading to sample degradation. We utilize a multifocal scanhead splitting a laser beam into a line of 64 foci, allowing sample illumination in real time at full laser power. This technology requires charge-coupled device field detection in contrast to conventional detection by photomultipliers. A comparison of the optical performance of both setups shows functional equivalence in every measurable parameter down to penetration depths of 200 microm, where most actual experiments are executed. The advantage of photomultiplier detection materializes at imaging depths >300 microm because of their better signal/noise ratio, whereas only charge-coupled devices allow real-time detection of rapid processes (here blood flow). We also find that the point-spread function of both devices strongly depends on tissue constitution and penetration depth. However, employment of a depth-corrected point-spread function allows three-dimensional deconvolution of deep-tissue data up to an image quality resembling surface detection.en
dc.language.isoenen
dc.subject.meshAnimalsen
dc.subject.meshBrainen
dc.subject.meshCalibrationen
dc.subject.meshCell Nucleusen
dc.subject.meshEquipment Designen
dc.subject.meshHippocampusen
dc.subject.meshImage Processing, Computer-Assisteden
dc.subject.meshLasersen
dc.subject.meshLighten
dc.subject.meshMiceen
dc.subject.meshMice, Transgenicen
dc.subject.meshMicroscopyen
dc.subject.meshMicroscopy, Confocalen
dc.subject.meshPhotonsen
dc.subject.meshSepharoseen
dc.titleThe power of single and multibeam two-photon microscopy for high-resolution and high-speed deep tissue and intravital imaging.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre for Infection Research, Junior Research Group Immunodynamics, D-38124 Braunschweig, Germany.en
dc.identifier.journalBiophysical journalen

Related articles on PubMed

All Items in HZI are protected by copyright, with all rights reserved, unless otherwise indicated.