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2015 ; 5
(ä): 18303
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Development of a real-time flexible multiphoton microendoscope for label-free
imaging in a live animal
#MMPMID26673905
Ducourthial G
; Leclerc P
; Mansuryan T
; Fabert M
; Brevier J
; Habert R
; Braud F
; Batrin R
; Vever-Bizet C
; Bourg-Heckly G
; Thiberville L
; Druilhe A
; Kudlinski A
; Louradour F
Sci Rep
2015[Dec]; 5
(ä): 18303
PMID26673905
show ga
We present a two-photon microendoscope capable of in vivo label-free deep-tissue
high-resolution fast imaging through a very long optical fiber. First, an
advanced light-pulse spectro-temporal shaping device optimally precompensates for
linear and nonlinear distortions occurring during propagation within the
endoscopic fiber. This enables the delivery of sub-40-fs duration infrared
excitation pulses at the output of 5?meters of fiber. Second, the endoscopic
fiber is a custom-made double-clad polarization-maintaining photonic crystal
fiber specifically designed to optimize the imaging resolution and the intrinsic
luminescence backward collection. Third, a miniaturized fiber-scanner of 2.2?mm
outer diameter allows simultaneous second harmonic generation (SHG) and
two-photon excited autofluorescence (TPEF) imaging at 8 frames per second. This
microendoscope's transverse and axial resolutions amount respectively to 0.8??m
and 12??m, with a field-of-view as large as 450??m. This microendoscope's
unprecedented capabilities are validated during label-free imaging, ex vivo on
various fixed human tissue samples, and in vivo on an anesthetized mouse kidney
demonstrating an imaging penetration depth greater than 300??m below the surface
of the organ. The results reported in this manuscript confirm that nonlinear
microendoscopy can become a valuable clinical tool for real-time in situ
assessment of pathological states.