Measurement of thickness and profile of a transparent material using fluorescent stereo microscopy.

Full-field thickness measurement for a thin transparent film is of interest for biological, medical, electronic, and packaging materials. It is a challenging task when the film is curvy, delicate and its thickness varies with location. We report herein a method to measure the thickness of a transparent (flat or curved) material and its topography using a stereo fluorescent profilometry technique. In this technique, two different types of fluorescent particles are deposited to both sides of the transparent film. Selected fluorescent excitation and emission are used to allow the observation of each one surface of the film at a time to determine the surface profile using stereo-based digital image correlation techniques. After the surface profiles for both surfaces are determined, subtraction of one surface profile from the other provides accurate thickness distribution of the film. Validation experiments were conducted using transparent films with known thickness. As an application, a measurement on a contact lens was conducted. The technique is appropriate for measurement of the full-field thickness of objects at other scales, such as soft transparent or translucent biofilms, with which thickness can hardly be measured accurately with other techniques.