Digital lensless holographic microscopy (DLHM) allows the design of cost-effective systems using off-the-shelf materials, making this type of lensless microscope accessible to many users worldwide. However, these materials may have a limited optomechanical performance that is aggravated due to the sought compactness and the intended cost-effective manufacturing process. This problem particularly affects the illumination source, which is of critical importance for DLHM, as it defines the optical performance of the microscope. While recent reports show that the required point source can be built from a low-cost laser diode coupled to an also low-cost aspheric lens, the resulting illumination has a distorted wavefront that limits the performance of the microscope. A simple homemade setup to correct the distortion of such illumination source and its integration into a compact, cost-effective, DIY, and open-source-certifiable digital lensless holographic microscope, is presented. The distortion-corrected DLHM is validated by imaging calibrated test targets and biological samples, achieving a 12-fold extension on the distortion-free magnification range of previous designs and a doubling of the effective spatial resolution without significant increments in its overall cost.
A multi-view occlusion removal method for digital lensless holographic microscopy (DLHM) is presented. Multiple DLHM holograms, whose individual reconstructions show occluded or partially occluded sample details, are recorded for different sample placements at its plane. A coordinated addition of the multiple DLHM recordings produces a composite hologram whose reconstruction allows the removal of the occlusions for a given imaging plane while increasing the reconstructed field of view. A theoretical model supports the method and its feasibility is tested with phase bio- and non-bio samples.
The light source in digital lensless holographic microscopy (DLHM) plays a key role in the optical performance of this microscopy methodology. The underlying physics that support DLHM rely on having an outward propagating spherical wavefront illuminating the sample to record on the surface of a digital camera the intensity of the diffracted wavefield that is magnified by free-space propagation; this first stage constitutes the recording. The information of the sample is numerically retrieved by computing the diffraction that an inward propagating spherical wavefront undergoes in the recorded intensity in a stage known as reconstruction. In both stages of this lensless microscopy method, it is assumed that perfect spherical wavefronts are utilized; hence, the closer the illumination light sources to exp[±ikr]/|r| are, the better the performance of the microscope. For the recovery of the information of the sample, the accuracy of the inward propagating spherical wavefront exp[−ikr]/|r| is guaranteed by its correct sampling in the numerical computation of the diffraction process; thus, the main challenge is the physical production of light sources to illuminate the sample whose amplitude profile is accurately described by exp[ikr]/|r|. In this work, the different approaches that our research group has explored to produce illuminating point sources for DLHM are evaluated. Point sources based on pinholes, coned-shaped optical fibers, optical pick-up units, and aspheric lenses are evaluated by rating their robustness, compactness, reliability, cost and easiness of production, and their performance on imaging test targets and biosamples. Some additional DLHM light sources reported in the literature are also briefly reviewed based on their available data.
Since the discovery of the wave behavior of light, diffraction has been a cornerstone in optics. The teaching of the diffraction theory has been usually done theoretically based on a mathematical approach that could hinder the understanding of the physical phenomena. In this work, the simplicity in the architecture of an accessible, cost-effective, and 3D-printed digital lensless holographic microscope is used as an educational tool to study the diffraction theory by providing experimental validations of the phenomena for undergraduate students. The recording and reconstruction steps of the lensless holographic technique take the students to the bidirectionally of the diffraction phenomena in a completely hands-on approach. The integration of the theory with an accessible experimental setup generates an innovative way of teaching the diffraction phenomena in a classroom.
In this work, the content for an undergraduate-level holography workshop is presented. The session is divided into two parts: an instructional section and a hands-on application activity supported by research-grade open-source software. The first section starts with a brief theoretical review of conventional imaging, interference, and diffraction as the underlying physical phenomena. These concepts are then used for the description of the recording, processing, and reconstruction stages of analog holography, emphasizing in each case the phenomenology rather than the mathematical framework. Finally, the translation of these stages to digital holography is presented, introducing the principles of digital recording and numerical reconstruction. The contents of the instructional section are then applied in a lecturer-guided activity, in which the participants generate a computational off-axis hologram and calculate its reconstruction. All the operations are performed using the “Numerical Propagation” plugin of the open-source software ImageJ. This research-grade software allows the modeling and manipulation of complex-valued wavefields from a user-friendly graphical interface. It thus allows the participants to recreate step-by-step the recording and reconstruction stages of the holographic process, while directly identifying when each physical phenomenon is at play. The proposed content can be implemented either as a stand-alone workshop or as an applied component of an undergraduate optics course.
A single-shot procedure to reduce speckle noise in numerically computed complex-valued wavefields is presented. The method is supported by the possibility of numerically producing multiple speckle realizations of a calculated complex-valued wavefield to reduce the speckle noise through a noncoherent superposition of the produced realizations. Although the method is applied to digital holographic microscopy, it could be utilized in other techniques where a numerical representation of the complex-valued wavefield of interest can be obtained. Experimental results with nonbiological and biological samples are presented to support the feasibility of the method.
In phase-shifting digital holographic microscopy (PS-DHM), the reconstructed phase map is obtained after processing several holograms of the same scene with a phase shift between them. Most reconstruction algorithms in PS-DHM require an accurate and known phase shift between the holograms, requirement that limits the PS-DHM applicability. This work presents an iterative-blind phase shift extraction method based on the demodulation of the different components of the holograms using three-frame holograms with arbitrary and unequal phase-shifts. Both simulated and experimental results demonstrate the goodness and feasibility of the proposed technique.
A self-contained platform for realistic modeling of digital holographic microscopy (DHM) is presented. Amplitude and phase samples, imaged in different architectures, can be modeled to produce numerical DHM holograms that include all the parameters that are present in real experiments, providing an accessible way for newcomers to have a first approach to this research field. The platform is based on considering the imaging arm of the DHM that produces the object wave as the result of the convolution process between the geometrical-optics image prediction of the sample with the point spread function introduced by diffraction. The DHM hologram is produced by the amplitude superposition of complex-valued object wave with a reference wave of arbitrary description. The sampling of the analytically produced DHM holograms is set from the input discretized image according to the specifications of the digital camera aimed to be used. The feasibility of the realistic platform is exemplified by contrasting the wrong results of a nonrealistic simulation with experimental results to show the need for using a complete realistic simulation like the one presented; further applications of the platform to numerical modeling speckle noise reduction over samples with controlled levels of roughness, and phase-shifting DHM techniques, are included.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.