Simultaneous imaging of multiple focal planes using a two-photon scanning microscope.
Despite all the advances in nonlinear microscopy, all existing instruments are constrained to obtain images of one focal plane at a time. In this Letter we demonstrate a two-photon absorption fluorescence scanning microscope capable of imaging two focal planes simultaneously. This is accomplished by temporally demultiplexing the signal coming from two focal volumes at different sample depths. The scheme can be extended to three or more focal planes.
Amir
W
W
Department of Physics, Colorado School of Mines, Golden 80401, USA.
Carriles
R
R
Hoover
E E
EE
Planchon
T A
TA
Durfee
C G
CG
Squier
J A
JA
D004867:Equipment Design; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional; D007546:Isoquinolines / Q000737:chemistry / Q000494:pharmacology; D008855:Microscopy, Electron, Scanning / Q000295:instrumentation* / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D008863:Microspheres; D055095:Optics and Photonics; D017785:Photons; D013053:Spectrophotometry; D013997:Time Factors
D007546:Isoquinolines; C017475:lucifer yellow