Freeform optical surface shapes have evolved from an intriguing optical design concept to a practical necessity for applications ranging from space and defense to consumer electronics. The demand for freeform solutions is no more strongly felt than in the development of digital immersive displays for augmented and mixed reality, where the combination of exceptionally high performance combines with ergonomic constraints of wearable interactive technologies. Some of these advanced designs call for diffraction-limited performance at large fields of view in off-axis orientations, often through conformal surfaces. Freeform optics are often the only way to correct the resultant aberrations, but their manufacture demands high-precision, high resolution surface figure metrology data such as can be obtained using coherence scanning interferometric microstitching (CSIM).
Near-eye display performance is usually summarized with a few simple metrics such as field of view, resolution, brightness, size, and weight, which are derived from the display industry. In practice, near-eye displays often suffer from image artifacts not captured in traditional display metrics. This work defines several immersive near-eye display metrics such as gaze resolution, pupil swim, image contrast, and stray light. We will discuss these metrics and their trade-offs through review of a few families of viewing optics. Fresnel lenses are used in most commercial virtual reality near-eye displays in part due to their light weight, low volume and acceptable pupil swim performance. However, Fresnel lenses can suffer from significant stray light artifacts. We will share our measurements of several lenses and demonstrate ways to improve performance. Smooth refractive lens systems offer the option for lower stray-light viewing but usually at the cost of a much larger size and weight in order to get to the same pupil swim performance. This can be addressed by using a curved image plane but requires new display technology. Polarization-based pancake optics is promising and can provide excellent image resolution and pupil swim performance within an attractive form-factor. This approach, however, generally results in low light efficiency and poor image contrast due to severe ghosting. We will discuss some of the main limitations of that technology.
Electro-Chemical Polishing is routinely used in the anodizing industry to achieve specular surface finishes
of various metals products prior to anodizing. Electro-Chemical polishing functions by leveling the
microscopic peaks and valleys of the substrate, thereby increasing specularity and reducing light scattering.
The rate of attack is dependent of the physical characteristics (height, depth, and width) of the microscopic
structures that constitute the surface finish. To prepare the sample, mechanical polishing such as buffing or
grinding is typically required before etching. This type of mechanical polishing produces random
microscopic structures at varying depths and widths, thus the electropolishing parameters are determined in
an ad hoc basis. Alternatively, single point diamond turning offers excellent repeatability and highly
specific control of substrate polishing parameters. While polishing, the diamond tool leaves behind an
associated tool mark, which is related to the diamond tool geometry and machining parameters. Machine
parameters such as tool cutting depth, speed and step over can be changed in situ, thus providing control of
the spatial frequency of the microscopic structures characteristic of the surface topography of the substrate.
By combining single point diamond turning with subsequent electro-chemical etching, ultra smooth
polishing of both rotationally symmetric and free form mirrors and molds is possible. Additionally,
machining parameters can be set to optimize post polishing for increased surface quality and reduced
processing times. In this work, we present a study of substrate surface finish based on diamond turning tool
mark spatial frequency with subsequent electro-chemical polishing.
Significant progress has been made in the application of optogenetic stimulation as a means to modulate and control
cellular functions within chemically-identical groups of cells. High resolution imaging can detect subtle morphological
(shape/refractive index) changes in cells subsequent to optogenetic stimulation. Invasive topographical measurement
methods such as mainstream AFM and other scanning probe techniques suffer from low temporal resolution and
restricted field of view, resulting in reduced throughput, even though these methods exhibit high sensitivity to
morphological changes. QPM, integrated with optogenetic stimulation incorporates a wide-field, label-free, non-invasive
optical imaging technique for all optical stimulation and detection with high spatial and temporal resolution. We
dynamically monitored phase of cells, sensitized with and without ChR2, using quantitative phase microscopy with and
without light stimulation. The variation of phase in optogenetically stimulated cells (expressing ChR2) was found to be
higher than that of the control cells. We report that our method could potentially evaluate effectiveness of various opsins
and stimulation parameters including cellular function under different physiological surroundings via spatiallymodulated
optogenetic stimulation and wide-field quantitative phase imaging.
Refractive index (RI) and its dispersion play a major role in interaction of electromagnetic wave with matter. Quantitative
phase imaging (QPI) has proven to be a useful tool to estimate the RI from the sample-induced phase delay measurement at
high spatio-temporal resolution. Here, we introduce near-infrared dispersive quantitative phase imaging (NIRD-QPI) of
microscopic objects. The setup uses a new geometry for quantitative phase microscopy by use of spatial frequency filtering
in Fourier plane. High resolution refractive index spectroscopic measurement over a range from 690 to 840nm in interval of
25nm is reported. This method could prove to be very useful for characterizing wide range of nano and biomaterials.
RBC has been shown to possess shape memory subsequent to
shear-induced shape transformation. However, this
property of RBC may not be generalized to all kinds of stresses. Here, we report our observation on the action of
radiation pressure forces on RBC's shape memory using optical manipulation and quantitative phase microscopy
(OMQPM). QPM, based on Mach-Zehnder interferrometry, allowed measurement of dynamic changes of shape of RBC
in optical tweezers at different trapping laser powers. In high power near-infrared optical tweezers (>200mW), the RBC
was found to deform significantly due to optical forces. Upon removal of the tweezers, hysteresis in recovering its
original resting shape was observed. In very high power tweezers or long-term stretching events, shape memory was
almost erased. This irreversibility of the deformation may be due to temperature rise or stress-induced phase
transformation of lipids in RBC membrane.
Atomic Force Microscope (AFM) imaging, due to the scanning method of recording, requires significant recording
time for examination of wide sample area. In contrast, digital holographic microscopy (DHM), owing to the wide-field
method, allows recording of the hologram in very fast rate which could be numerically analyzed to reveal surface of the
sample with axial resolution at the nanometer scale. However, DHM yields quantitative phase properties of the sample,
and therefore sensitive to changes in refractive index along with physical thickness. Therefore, to accurately determine
the refractive index map, it is imperative to estimate the physical thickness map of the sample. This was achieved by
AFM imaging. Further, since the transverse resolution of DHM is limited by diffraction limit, co-registration of AFM
image provided higher transverse resolution at nanometer scale. The interference of the AFM probe was observed to be
minimal during simultaneous AFM and DHM recording due to the transparent nature and bent configuration of the
optical fiber based AFM cantilever. Integration of DHM and AFM led to realization of a powerful platform for
nanoscale imaging. The integrated AFM-DHM system was built on an inverted fluorescence microscope to enable
fluorescence imaging of the sample. The integrated system was employed to analyze fluorescent polystyrene
microspheres, two-photon polymerized microstructures and red blood cells.
Red blood cells (RBC) possess unique viscoelastic characteristics which allow them to pass through capillaries
narrower than their size. Measurement of viscoelastic property of cells (e.g. RBC) in low-force regime is of high
significance as it represents conditions of membrane fluctuation in response to physiological conditions.
Estimation of visco-elastic properties of RBC requires measurement of extent of deformation in RBC subjected
to known force. Optical tweezers, being gentle and absolutely sterile, are emerging as the tool of choice for
application of localized force on cells. However, stretching of RBC in very low force regime has not been
quantified. Further, though deformations in transverse directions have been measured, vertical deformations due
to stretching of cells cannot be quantified by classical microscopic images. Here, we report realization of offaxis
digital holographic microscopy (DHM) for highly sensitive axial changes in RBC shape due to stretching
by optical tweezers without attaching microscopic beads. The RBC was stretched in axial direction with
nanometer precision by change of divergence of the trapping beam. The obtained deformation patterns were
compared with the axial position of the tweezers focus. Since the pathophysiology of progression of diseases
like malaria and cancer is reflected in the biophysical (both mechanical and material) properties of the cells, it is
possible to identify the changes by simultaneous measurement of refractive index and elasticity using this
approach.
High throughput analysis of trapped samples requires effective loading and unloading into the trap in a microfluidic
environment. We demonstrate development of a hybrid optical transport trap (HOTT) which combines a tapered fiberoptic
2D trap for transport of microscopic objects into and out of the optical tweezers trap in an orthogonal geometry.
For small cone angle of the tip, the microscopic objects (polystyrene and red blood cells) were found to be trapped in
two-dimensions and pushed along the axial direction by domination of scattering force. This was found to be in
consistence with the estimated axial forces caused by the beam profiles emerging from the small-cone tapered fiber tip.
While for loading of the microscopic objects into the optical tweezers trap, the fiber tip was placed ~ 30μm away from
the tweezers trap, unloading was carried out in presence of the tip close (<15 μm) to the tweezers trap. Further, for a
fixed fiber trap and tweezers separation (~ 30 μm), both loading and unloading could be achieved by reducing the
tweezers trap power so that the scattering force exerted by the fiber trap exceeded the transverse gradient force of
tweezers trap. Since the tapered tip can be easily integrated onto a microfluidic channel, the proposed configuration can
find potential applications in lab-on-a-chip devices. We demonstrate analysis of transported microscopic objects using
digital holographic microscopy integrated with the HOTT.
While optical tweezers have been widely used for the manipulation and organization of microscopic objects in three
dimensions, observing the manipulated objects along axial direction has been quite challenging. In order to visualize
organization and orientation of objects along axial direction, we report development of a Digital holographic microscopy
combined with optical tweezers. Digital holography is achieved by use of a modified Mach-Zehnder interferometer with
digital recording of interference pattern of the reference and sample laser beams by use of a single CCD camera. In this
method, quantitative phase information is retrieved dynamically with high temporal resolution, only limited by frame
rate of the CCD. Digital focusing, phase-unwrapping as well as online analysis and display of the quantitative phase
images was performed on a software developed on LabView platform. Since phase changes observed in DHOT is very
sensitive to optical thickness of trapped volume, estimation of number of particles trapped in the axial direction as well
as orientation of non-spherical objects could be achieved with high precision. Since in diseases such as malaria and
diabetics, change in refractive index of red blood cells occurs, this system can be employed to map such disease-specific
changes in biological samples upon immobilization with optical tweezers.
Interaction of red blood cells (RBC) with optical tweezers has been found to differ under varied physiological and
pathological conditions as compared to its normal conditions. Earlier, we reported difference in rotation of trapped RBC
in hypertonic conditions for detection of malaria infection. Disk-like RBC when trapped in optical tweezers get oriented
in the vertical plane to maximize interaction with trapping beam. However, classical bright field, phase contrast or epifluorescence
microscopy cannot confirm its orientation, thus leading to ambiguous conclusions such as folding of RBC
during trapping by some researchers. Now, with use of digital holographic microscopy (DHM), we achieved high axial
sensitivity that confirmed orientation of trapped red blood cell. Further, DHM enabled quantitative phase imaging of
RBC under hypertonic condition. Dynamic changes of rotating RBC under optical tweezers at different trapping laser
power were evaluated by the use of DHM. The deviation from linear dependence of rotation speed of RBC on laser
power, was attributed towards deformation of RBC shape due to higher laser power (or speed).
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