Interferometric backward third harmonic generation microscopy for axial imaging with accuracy beyond the diffraction limit.
A new nonlinear microscopy technique based on interference of backward-reflected third harmonic generation (I-THG) from multiple interfaces is presented. The technique is used to measure height variations or changes of a layer thickness with an accuracy of up to 5 nm. Height variations of a patterned glass surface and thickness variations of fibroblasts are visualized with the interferometric epi-THG microscope with an accuracy at least two orders of magnitude better than diffraction limit. The microscopy technique can be broadly applied for measuring distance variations between membranes or multilayer structures inside biological tissue and for surface height variation imaging.
Department of Physics and Institute for Optical Sciences, University of Toronto, Toronto, Ontario, Canada; Department of Chemical and Physical Sciences, University of Toronto Mississauga, Mississauga, Ontario, Canada.
Daaf
D
Sandkuijl
Department of Physics and Institute for Optical Sciences, University of Toronto, Toronto, Ontario, Canada; Department of Chemical and Physical Sciences, University of Toronto Mississauga, Mississauga, Ontario, Canada.
Lukas
L
Kontenis
Matrix Dynamics Group, University of Toronto, Toronto, Ontario, Canada.
Nuno M
NM
Coelho
Matrix Dynamics Group, University of Toronto, Toronto, Ontario, Canada.
Christopher
C
McCulloch
Department of Physics and Institute for Optical Sciences, University of Toronto, Toronto, Ontario, Canada; Department of Chemical and Physical Sciences, University of Toronto Mississauga, Mississauga, Ontario, Canada.
Virginijus
V
Barzda
D000818:Animals; D002448:Cell Adhesion; D005347:Fibroblasts / Q000166:cytology; D005353:Fibronectins / Q000737:chemistry; D005898:Glass / Q000737:chemistry; D006801:Humans; D051379:Mice; D008857:Microscopy, Interference / Q000379:methods*; D041681:NIH 3T3 Cells
D005353:Fibronectins