title: Highly confined surface imaging by solid immersion total internal reflection fluorescence microscopy.
abstract: We report the use of a high-refractive-index aplanatic solid immersion lens (ASIL) in total internal reflection fluorescence (TIRF) microscopy. This new solid immersion total internal reflection fluorescence (SITIRF) microscopy allows highly confined surface imaging with a significantly reduced imaging depth compared with conventional TIRF microscopy. We explore the application of a high refractive index, low optical dispersion material zirconium dioxide in the SITIRF microscope and also introduce a novel system design which enables the SITIRF microscope to work either in the epi-fluorescence or TIRF modes with variable illumination angles. We use both synthetic and biological samples to demonstrate that the imaging depth in the SITIRF microscope can be confined to a few tens of nanometers. SITIRF microscopy has the advantages of performing highly selective imaging and high-resolution high-contrast imaging. Potential applications in biological imaging and future developments of SITIRF microscopy are proposed.
authors.affiliation: Department of Physics and Astronomy, The University of Sheffield, The Hicks Building, Hounsfield Road, Sheffield, S3 7RH, UK. l.wang@sheffield.ac.uk
authors.forename: Lin
authors.initials: L
authors.lastname: Wang
authors.forename: Cvetelin
authors.initials: C
authors.lastname: Vasilev
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Canniffe
authors.forename: Luke R
authors.initials: LR
authors.lastname: Wilson
authors.forename: C Neil
authors.initials: CN
authors.lastname: Hunter
authors.forename: Ashley J
authors.initials: AJ
authors.lastname: Cadby
mesh_terms: D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000295:instrumentation*; D007909:Lenses; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods*