Multiphoton fluorescence excitation: new spectral windows for biological imaging

Chris Xu; Jerome Mertz; J. B. Shear; Watt W. Webb

doi:10.1117/12.258073

13 November 1996 Multiphoton fluorescence excitation: new spectral windows for biological imaging

Chris Xu, Jerome Mertz, J. B. Shear, Watt W. Webb

Proceedings Volume 2819, Imaging Spectrometry II; (1996) https://doi.org/10.1117/12.258073
Event: SPIE's 1996 International Symposium on Optical Science, Engineering, and Instrumentation, 1996, Denver, CO, United States

Abstract

Nonlinear excitation of fluorophores through molecular absorption of two or three near infrared photons from the tightly focused femtosecond pulses of a mode-locked laser offers the cellular biologist an unprecedented panoply of biomolecular indicators for microscopic imaging and cellular analysis. Measurements of the two-photon excitation spectra of more than twenty ultra-violet and visible absorbing fluorophores from 690 to 1050 nm reveal useful cross sections for near infrared excitation, providing an artist's palette of emission markers and chemical indicators for living biological preparations. Measurements of three-photon fluorophore excitation spectra now define alternative windows of relatively benign wavelength to excite deeper UV fluorophores. The three-photon excitation spectrum of the amino acid tryptophan, measured 700-900 nm, delivers native fluorescence for imaging and assay of proteins and neurotransmitter sin living tissues. The inherent optical sectioning capabilities of focused nonlinear excitation provides 3D resolution for imaging and avoids out of focus background. Here, we describe the characteristics of the measured nonlinear excitation spectra and define the resulting features of nonlinear microscopy for biological imaging.

Citation Download Citation

Chris Xu, Jerome Mertz, J. B. Shear, and Watt W. Webb "Multiphoton fluorescence excitation: new spectral windows for biological imaging", Proc. SPIE 2819, Imaging Spectrometry II, (13 November 1996); https://doi.org/10.1117/12.258073

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
3 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Luminescence

Microscopy

Multiphoton fluorescence microscopy

Multiphoton microscopy

Photons

Confocal microscopy

Ultraviolet radiation

Show All Keywords

Keywords/Phrases

Search In:

Publication Years