Nonlinear optical potentiometric dyes optimized for imaging with 1064-nm light.
Nonlinear optical phenomena, such as two-photon fluorescence (2PF) and second harmonic generation (SHG), in combination with voltage sensitive dyes, can be used to acquire high-resolution spatio temporal maps of electrical activity in excitable cells and tissue. Developments in 1064-nm fiber laser technology have simplified the generation of high-intensity, long-wavelength, femtosecond light pulses, capable of penetrating deep into tissue. To merge these two advances requires the design and synthesis of new dyes that are optimized for longer wavelengths and that produce fast and sensitive responses to membrane potential changes. In this work, we have systematically screened a series of new dyes with varying chromophores and sidechains that anchor them in cell membranes. We discovered several dyes that could potentially be used for in vivo measurements of cellular electrical activity because of their rapid and sensitive responses to membrane potential. Some of these dyes show optimal activity for SHG; others for 2PF. This regulated approach to dye screening also allows significant insight into the molecular mechanisms behind both SHG and 2PF. In particular, the differing patterns of sensitivity and kinetics for these two nonlinear optical modalities indicate that their voltage sensitivity originates from differing mechanisms.
Teisseyre
Thomas Z
TZ
University of Connecticut Health Center, Richard D. Berlin Center for Cell Analysis and Modeling, 263 Farmington Avenue, MC-1507, Farmington, Connecticut 06030-1507, USA.
Millard
Andrew C
AC
Yan
Ping
P
Wuskell
Joseph P
JP
Wei
Mei-de
MD
Lewis
Aaron
A
Loew
Leslie M
LM
D000200:Action Potentials / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D005456:Fluorescent Dyes / Q000737:chemistry; D007089:Image Enhancement / Q000379:methods*; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D017711:Nonlinear Dynamics; D055095:Optics and Photonics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
D005456:Fluorescent Dyes