Abstract
We report combined temporal and spatial laser pulse shaping to perform lateral and depth dependent two-photon excited fluorescence of dyes. For generating the specific spatially and temporally phase tailored pulses a temporal pulse shaper and a subsequent spatial pulse shaper are employed. Simultaneous spatial and temporal shaping is presented for two-photon excited fluorescence by applying temporal third order phase functions on spatially different light field components. Moreover, the prospects of spatial shaping are demonstrated by applying various lateral two-photon fluorescence pattern. In particular, a depth dependent excitation of different dyes is performed which leads to a high axially resolved fluorescence contrast. The introduced spatial and temporal shaping technique provides new perspectives for biophotonic imaging applications.
Highlights
In the last years laser pulse shaping has become a powerful and versatile tool in several research fields (e.g. [1,2,3,4,5,6])
We report combined temporal and spatial laser pulse shaping to perform lateral and depth dependent two-photon excited fluorescence of dyes
Simultaneous spatial and temporal shaping is presented for two-photon excited fluorescence by applying temporal third order phase functions on spatially different light field components
Summary
In the last years laser pulse shaping has become a powerful and versatile tool in several research fields (e.g. [1,2,3,4,5,6]). Similar to the temporal shaping it is possible to modify the spatial laser profile by using focussing lenses and a two dimensional liquid crystal array combined to a spatial light shaper [13, 14] This wavefront shaping technique leads to definable tailored beam profiles in the focal plane. The combined shaping method is applied to first establish depth dependent selective excitation of different dyes, which leads to a high axially resolved fluorescence contrast. These optical tools will be relevant for biophotonic imaging
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