Optimizing the fluorescent yield in two-photon laser scanning microscopy with dispersion compensation

Opt Express. 2010 Jun 21;18(13):13661-72. doi: 10.1364/OE.18.013661.


A challenge for nonlinear imaging in living tissue is to maximize the total fluorescent yield from each fluorophore. We investigated the emission rates of three fluorophores-rhodamine B, a red fluorescent protein, and CdSe quantum dots-while manipulating the phase of the laser excitation pulse at the focus. In all cases a transform-limited pulse maximized the total yield to insure the highest signal-to-noise ratio. Further, we find evidence of fluorescence antibleaching in quantum dot samples.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Biology / instrumentation
  • Cadmium Compounds / chemistry
  • Luminescent Proteins / chemistry*
  • Microscopy, Confocal / methods*
  • Microscopy, Fluorescence / methods*
  • Models, Theoretical
  • Photobleaching
  • Plant Cells
  • Quantum Dots*
  • Rhodamines / chemistry*
  • Selenium Compounds / chemistry


  • Cadmium Compounds
  • Luminescent Proteins
  • Rhodamines
  • Selenium Compounds
  • red fluorescent protein
  • cadmium selenide
  • rhodamine B