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2016 ; 7
(10
): 3958-3967
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Scanless functional imaging of hippocampal networks using patterned two-photon
illumination through GRIN lenses
#MMPMID27867707
Moretti C
; Antonini A
; Bovetti S
; Liberale C
; Fellin T
Biomed Opt Express
2016[Oct]; 7
(10
): 3958-3967
PMID27867707
show ga
Patterned illumination through the phase modulation of light is increasingly
recognized as a powerful tool to investigate biological tissues in combination
with two-photon excitation and light-sensitive molecules. However, to date
two-photon patterned illumination has only been coupled to traditional microscope
objectives, thus limiting the applicability of these methods to superficial
biological structures. Here, we show that phase modulation can be used to
efficiently project complex two-photon light patterns, including arrays of points
and large shapes, in the focal plane of graded index (GRIN) lenses. Moreover,
using this approach in combination with the genetically encoded calcium indicator
GCaMP6, we validate our system performing scanless functional imaging in rodent
hippocampal networks in vivo ~1.2 mm below the brain surface. Our results open
the way to the application of patterned illumination approaches to deep regions
of highly scattering biological tissues, such as the mammalian brain.