TY - JOUR
T1 - Non-scanning motionless fluorescence three-dimensional holographic microscopy
AU - Rosen, Joseph
AU - Brooker, Gary
N1 - Funding Information:
This work was supported by National Science Foundation grant no. 0420382 and CellOptic. We thank M. DeBernardi, B. Storrie and S. Krueger for valuable comments and A. Hermerschmidt, who provided the special firmware for the SLM. We also thank W.R. Kennedy and G. Wendelschager-Crabb for the immunolabelled slide of skin and K. Ryan for deconvolving the stack of images created from the hologram. Correspondence should be addressed to G.B. Supplementary information accompanies this paper on www.nature.com/naturephotonics.
PY - 2008/3/1
Y1 - 2008/3/1
N2 - Holography is an attractive imaging technique as it offers the ability to view a complete three-dimensional volume from one image. However, holography is not widely applied to the field of three-dimensional fluorescence microscopic imaging, because fluorescence is incoherent and creating holograms requires a coherent interferometer system. Although scanning one beam of an interferometer pattern across the rear aperture of an objective to excite fluorescence in a specimen overcomes the coherence limitation, the mechanical scanning is complicated, which makes the image capturing slow, and the process is limited to low-numerical-aperture objectives. Here we present the first demonstration of a motionless microscopy system (FINCHSCOPE) based on Fresnel incoherent correlation holography, and its use in recording high-resolution three-dimensional fluorescent images of biological specimens. By using high-numerical-aperture objectives, a spatial light modulator, a CCD camera and some simple filters, FINCHSCOPE enables the acquisition of three-dimensional microscopic images without the need for scanning.
AB - Holography is an attractive imaging technique as it offers the ability to view a complete three-dimensional volume from one image. However, holography is not widely applied to the field of three-dimensional fluorescence microscopic imaging, because fluorescence is incoherent and creating holograms requires a coherent interferometer system. Although scanning one beam of an interferometer pattern across the rear aperture of an objective to excite fluorescence in a specimen overcomes the coherence limitation, the mechanical scanning is complicated, which makes the image capturing slow, and the process is limited to low-numerical-aperture objectives. Here we present the first demonstration of a motionless microscopy system (FINCHSCOPE) based on Fresnel incoherent correlation holography, and its use in recording high-resolution three-dimensional fluorescent images of biological specimens. By using high-numerical-aperture objectives, a spatial light modulator, a CCD camera and some simple filters, FINCHSCOPE enables the acquisition of three-dimensional microscopic images without the need for scanning.
UR - http://www.scopus.com/inward/record.url?scp=40249118456&partnerID=8YFLogxK
U2 - 10.1038/nphoton.2007.300
DO - 10.1038/nphoton.2007.300
M3 - Article
AN - SCOPUS:40249118456
SN - 1749-4885
VL - 2
SP - 190
EP - 195
JO - Nature Photonics
JF - Nature Photonics
IS - 3
ER -