The power of single and multibeam two-photon microscopy for high-resolution and high-speed deep tissue and intravital imaging.
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
MetadataShow full item record
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.
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.
AffiliationHelmholtz Centre for Infection Research, Junior Research Group Immunodynamics, D-38124 Braunschweig, Germany.
- Intravital two-photon microscopy: focus on speed and time resolved imaging modalities.
- Authors: Niesner RA, Andresen V, Gunzer M
- Issue date: 2008 Feb
- Resolution enhancement of two-photon microscopy via intensity-modulated laser scanning structured illumination.
- Authors: Yeh CH, Chen SY
- Issue date: 2015 Mar 20
- An evaluation of two-photon excitation versus confocal and digital deconvolution fluorescence microscopy imaging in Xenopus morphogenesis.
- Authors: Periasamy A, Skoglund P, Noakes C, Keller R
- Issue date: 1999 Nov 1
- Evaluation of confocal microscopy system performance.
- Authors: Zucker RM, Price O
- Issue date: 2001 Aug 1
- Two-photon excitation of fluorescence for three-dimensional optical imaging of biological structures.
- Authors: Diaspro A, Robello M
- Issue date: 2000 Mar