A multifocal multiphoton volumetric imaging approach for high speed time-resolved FRET imaging in vivo

In this paper we will discuss the development of a
multifocal multiphoton fluorescent lifetime imaging
system where 4 individual fluorescent intensity and
lifetime planes are acquired simultaneously, allowing us
to obtain volumetric data without the need for sequential
scanning at different axial depths. Using a phase-only
SLM with an appropriate algorithm to generate a
holographic pattern, we project a beamlet array within a
sample volume of a size, which can be pre-programmed
by the user. We demonstrate the capabilities of the
system to image live-cell interactions. Whilst only 4
planes are shown, this technique can be rescaled to a
large number of focal planes enabling full 3D acquisition
and reconstruction.