Optimizing Multi-photon Fluorescence 

Microscopy Light Collection by Total 

Emission Detection (TED)

Christian Combs, Jay Knutson, Robert S. 

Balaban

National Heart Lung and Blood Institute

National Institutes of Health

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National Institutes of Health
• Multi-photon fluorescence microscopy (MPFM) is an 

imaging technique that can investigate biological 

processes to sub-cellular resolution at depths of hundreds 

of microns below the surface of biological tissues. 

• MPFM is currently limited by the use of inefficient light 

collection systems—detects a fraction of the light emitted 

from sample 

Technology

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New system maximizes probability of collecting all 

emitted fluorescent light to a detector

• Enhanced brightness of light from the sample

• Increase signal-to-noise ratio (SNR). 

• Improved time resolution

• Reduced laser power requirements

• Reduced photodynamic damage

• Stage of research: Prototype

• PCT Application #PCT/US2007/17478 filed 08/06/2007 

US Application #11/979,600 filed 11/06/2007 

Total Emission Detection (TED)

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Total Emission Detection: Concept

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TED II for in vivo Studies

Design: Jay Knutson

Built by: Brad Bobbitt 

of LSM Tech.

DET

2

from

IR

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Parabola Collected Light Objective Collected Light

TED II collects 2.5x more light than objective alone

DS-red tagged actin in blood vessel of 

intact mouse brain

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Optimized MPFM emission detection of deep 

fluorescence imaging of biological tissues and cells

Commercial Applications

Licensing Status: Available for licensing

Collaborative Research Opportunity:

Further develop, evaluate, or commercialize a total 

emission detection system for multi-photon imaging.

Collaboration Opportunities

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National Institutes of Health
Collaborators and Contributors to This Work

• Dr. Robert Balaban and Dr. Merav Lugar-Hammer 

(NHLBI Lab of Cardiac Energetics)

• Drs. Jay Knutson and Aleksander Smirnov

(NHLBI Lab of Biophysical Chemistry)

• Drs. Amir Gandjbakhche and Jason Riley

(NICHD Section on Biomedical Stochastic Physics) 

• Dr. Christian A. Combs

(NHLBI Light Microscopy Core Facility)

• Brad Bobbitt

(LSM Technologies, Stewartstown, PA)

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Contact Information

For further information contact:

Licensing:

Michael Shmilovich Esq.

NIH Office of Technology Transfer

shmilovm@mail.nih.gov

301-435-5019

Collaboration:

Alan Deutch, Ph.D.

NHLBI Office of Technology Transfer and Development

deutcha@nhlbi.nih.gov

301-402-5579

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National Institutes of Health