High Magnification Microscopic System Focusing on Transparent Liquid/Liquid Interface Formed in Thin-Layer Two-Phase Microcell

Abstract

A thin-layer two-phase microcell was fabricated without an adhesive, which showed no fluorescence and resisted acids. The lower hollow of the microcell was thin (0.18 mm), which was in the range of the working distance of the used objective of high magnification (60×) and high numerical aperture (1.2). A reflection method using probe light was newly employed for focusing the objective on a transparent dodecane/water interface strictly with an inverted microscope. The system was applied to in situ fluorescence microscopic measurements of the dynamics of single DNA molecules (165600 base pairs) at the dodecane/water interface.