We report the development of the Three-dimensional Real-time Uninvasive Imaging and Evaluation (TRUImagE) system
based on digital holographic microscopy to study the morphological changes in cells undergoing photodynamic therapyinduced
cell death. The optical system, based on the Michelson interferometer and configured in transmission mode, and
the sample holder incorporating a stage incubator have been developed for monitoring various tumorigenic cell samples
without the use of markers. Off-axis digital holograms were recorded with a CCD sensor and numerically reconstructed
to provide quantitative phase imaging and 3D morphology of the cells in real time. The system was used to continuously
monitor and study, at different time points, the changes in cells after incubation with the photosensitizer followed by
activation by the appropriate light dose. Results obtained from the TRUImagE system and biochemical assays will be
given.
Micro optics characterization by use of digital holographic microscopy (DHM) is proposed recently. DHM can provide
phase image and very suitable for the quantitative mapping of transmission material with a certain refractive index.
However, it has been found that in DHM the microscope objective introduces a spherical phase curvature to the object
wave which may disturb the measurement especially for the micro-lens array. We present single lens characterization
and uniformity inspection of micro-lens array by use of a new concept DHM system developed recently. The new
concept DHM is based on a single cube beam-splitter (SCBS) configuration using an MO to provide high resolution on
the test specimen. The SCBS is put into the optical path with a small angle between the optical axis and its central semireflecting
layer. In this way, light is split into two parts when in and combined to form two holograms when out of SCBS.
For the symmetrical configuration of the beam splitter cube, the spherical phase curvature introduced by the MO can be
physically compensated during interference. Because no separated light propagation outside the SCBS, the whole system
is insensitive to vibration. As light coming out of the MO serves not only the object beam but also the reference beam, it
enables the inspection of the uniformity across a whole micro-lens array. Geometrical characterisation of the shape and
surface roughness of micro-lens is given as well as the uniformity analysis across the whole array.
The inspection and characterisation of laser marks using digital holographic microscopy (DHM) is presented in this
paper. A DHM system in transmission mode was designed and the reconstruction algorithm for this configuration was
investigated. The software was developed to provide live reconstruction of holograms for real time numerical evaluation
of amplitude and phase contrast images. A CO2 laser-based marking system was employed to create marks on glass
substrates. By analysing the quality, 3D profile measurement, and material distribution of the marked area, the
parameters of the laser system could be optimised to achieve the desired mark. The phase contrast images provide
quantitative refractive index analysis and 3D profile studies. The results were compared with those obtained using white
light confocal microscopy. The capabilities and advantages of the DHM system for the analysis of laser marks are also
presented.
KEYWORDS: Digital holography, Holograms, Holography, Microscopes, 3D metrology, 3D image processing, Reflection, 3D image reconstruction, Glasses, Silicon
We report the development of a simple commercial digital holographic microscope. The hologram is
recorded using a CCD sensor and numerically reconstructed to provide quantitative analysis of the
object. The laser source is coupled via fibre optics and the opto-mechanical setup is flexible and
customizable for either the reflection or transmission mode. The user-friendly software allows live
reconstruction, simultaneously providing both the amplitude and phase images. System performance is
improved with phase unwrapping and interferometric comparison. Additional features include various
image enhancements, cross-sectional and line profiling, measurement and data analysis tools for
quantitative 3D imaging and surface topography measurement. The performance of the product is
tested on different micro devices, glass and silicon surfaces.
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