Non-Contact Total Emission Detection Methods for Multiphoton Microscopy: Improved Image Fidelity and Biological Sample Analysis

The technology offered for licensing and for further development is in the field of multiphoton microscopy (MPM). More specifically, the invention describes and claims optical designs that can enhance and extend the capabilities of MPM in spectral imaging of biological samples. The unique design of the light collection and the detection optics maximizes the collection of emitted light, thus increasing the signal and hence the signal-to-noise ratio (SNR). Improvement in image fidelity will result in improved analysis of biological samples and thus will favorably impact medical research and possibly clinical diagnosis. The present technology is a further improvement on the TED (Total Emission Detection) technology, first disclosed by Dr. Robert Balaban et al. at the NIH in 2006 and claimed in US patent application 11/979,600, now allowed (Patent Publication US-2008-0063345 A1, March 13, 2008). The earlier NIH TED technology proposed an optical design based on enveloping the entirety of a small sample in a parabolic mirror/condenser combination so light emanated by a sample in all directions is redirected to the detector. The present technology further expands the capabilities of TED as its unique design employing parabolic, toric and conic mirrors ensures maximum light collection from large samples in cases where there is only access to one side of the tissues (e.g., in vivo or ex vivo). This is accomplished by the redirection of all attainable light (i.e., light escaping the tissue or a whole animal in the epi and sideway directions) to the detector.

Potential Commercial Applications: Competitive Advantages:
  • Tissue and cell analysis in biomedical research
  • Potential applications in clinical diagnostics
  The advent of multiphoton microscopy (MPM) provided several advantages in comparison to single-photon confocal microscopy. In particular the nonlinear optics used with this technology, combined with the elimination of a confocal pinhole aperture, led to direct sectioning and the use of lower energy photons. This approach preserves the integrity of the observed object (i.e. tissue) thus improving imaging results. The technology presented here further enhances the capabilities of MPM by providing the following advantages:
  • Increased signal-to-noise ratio
  • Enhanced image resolution due to SNR
  • Improved analytical capabilities
  • Non-contact
  • May readily be adaptable to commercial microscopes

Jay Knutson (NHLBI)

Intellectual Property:
PCT Application No. PCT/US2010/041723
US Application No. 61/224,772
US Application No. 13/383,248

US Patent Application Publication US-2008-0063345 A1, March 13, 2008.
Presentation, 7th EBSA European Biophysics Congress, July 11-15 2009, Genova, Italy (
CA Combs, AV Smirnov, JD Riley, AH Gandjbakhche, JR Knutson, RS Balaban. Optimization of multiphoton excitation microscopy by total emission detection using a parabolic light reflector. J Micros. 2007 Dec;228(Pt3):330-337. PubMed: 18045327

Collaboration Opportunity:

The NHLBI Laboratory of Molecular Biophysics is seeking statements of capability or interest from parties interested in collaborative research to further develop, evaluate, or commercialize an enhanced method of multiphoton microscopy that is suitable for the spectral imaging of biological samples. Please contact Brian W. Bailey, Ph.D. at for more information.

Licensing Contact:
Michael Shmilovich , Esq.
NIH Office of Technology Transfer
Phone: 301-435-5019

OTT Reference No: E-236-2009/0

Updated: Dec-28-2009