Home Personnel Research Application Notes Collaborations Hardware Software Publications 2PE Cross-sections Contact
 
   

 

An NIH-NIBIB Biomedical Technology Resource

 

 

 

Research

Project I - Technology Development for Improved Intravital Multiphoton Microscopy

 

This page is currently under construction.

 

 

 

 

 

 

 

 

 

 

 

 

A

 

 

We are using mouse peritoneum as a model tissue because it is easy to handle and represents a tissue that is relatively difficult to image through. We have constructed a 2f system in which the phase modulator is optically conjugate to the back aperture of a 20x/0.5 NA dipping objective. The PSF from this objective (fluorescent spot in a solution of fluorophore) is then imaged from the side through a coverslip using a 50x/0.55 NA Mitutoyo objective with a 13mm working distance (Figure. I-18). This strategy allows us to directly view aberrations and the effects on them by introduced phase profiles.  Currently, the wavefront surface and its 2p-modulo counterpart are controlled by algorithms written in the IDL image analysis environment. These algorithms will be converted to C/C++ for faster operation necessary for live specimens.  Correction phase profiles are calculated using a simple algorithm that steps through Zernike modes and optimizes for the maximum integrated value from the imaged PSF.  Preliminary results suggest that the PSF can be relatively well corrected through peritoneum (Figure I-18).  We will use this system initially to characterize the types of aberrations that are introduced by a variety of biological tissues.  

 

Enlarge This Image

 

 

References

Beaurepaire, E. and J. Mertz (2002). "Epifluorescence collection in two-photon microscopy." Appl. Opt. 41(25): 5376-5382.


Benninger, R., W. Ashby, E. A. Ring and P. W. Piston (2008). "Single-photon-counting detector for increased sensitivity in two-photon laser scanning microscopy." OPTICS LETTERS 33(24): 2895-2897.


Booth, M. J., M. A. A. Neil and T. Wilson (1998). "Aberration correction for confocal imaging in refractive-index-mismatched media." Journal of Microscopy-Oxford 192: 90-98.


Bor, Z. (1989). "Distortion of femtosecond laser pulses in lenses." Optics Letters 14(2): 119-121.


Burns, A., P. Sengupta, T. Zedayko, B. Baird and U. Wiesner (2006). "Core/shell fluorescent silica nanopartictes for chemical sensing: Towards single-particle laboratories." Small 2(6): 723-726.


. G., M. L. Ellsworth and R. N. Pittman (1990). "Determination of red blood cell oxygenation in vivo by dual video densitometric image analysis." Am J Physiol 258(4 Pt 2): H1216-23.


Fork, R. L., O. E. Martinez and J. P. Gordon (1984). "Negative Dispersion Using Pairs of Prisms." Optics Letters 9(5): 150-152.



Patterson, G. H. and D. W. Piston (2000). "Photobleaching in two-photon excitation microscopy." Biophys J 78(4): 2159-62.


Planchon, T. A., S. Ferre, G. Hamoniaux, G. Cheriaux and J. P. Chambaret (2004). "Experimental evidence of 25-fs laser pulse distortion in singlet beam expanders." Opt Lett 29(19): 2300-2.


Prieto, P. M., E. J. Fernandez, S. Manzanera and P. Artal (2004). "Adaptive optics with a programmable phase modulator: applications in the human eye." Optics Express 12(17): 4059-4071.


Rueckel, M., J. A. Mack-Bucher and W. Denk (2006). "Adaptive wavefront correction in two-photon microscopy using coherence-gated wavefront sensing." Proc Natl Acad Sci U S A 103(46): 17137-42.


Shapiro, I. M. and V. Srinivas (2007). "Metabolic consideration of epiphyseal growth: survival responses in a taxing environment." Bone 40(3): 561-7.


 

 



: Lab Schedule

: Group Presentations

: Protocols


- Biomedical Engineering
- Applied & Eng. Physics
- College of Engineering
- Cornell University

- Bio-Imaging Seminar
- Biophysics Seminar

- MPM Documentation
- DRBIO MPM Blog
- DRBIO Subversion

 



Search Google
DRBIO

 

Home · Personnel · Research · Application Notes · Collaborations · Hardware · Software · Publications · 2PE Cross Sections