Digital holographic microscopy (DHM) is a technique that has high potential for analyzing biological samples and has been successfully applied to the study of cells and cell lines providing information about important parameters such as refractive index, morphology, and dry mass, among others; it has also found applicability to study the effects of therapeutic treatments. Finding the size and shape of cells is important since they tend to change in the presence of some pathologies. In this research work, we obtain the morphology thickness and refractive index of the A375 melanoma cell line through a slight tilting of the cell in a DHM setup. Further, the development of a novel mathematical expression based on this tilt and in the optical phase difference is presented. We show images of melanoma cells with the refractive index information included, and their morphology thickness as rendered from the holographic phase maps recorded with DHM.

Current industrial demand for optical nondestructive testing includes the displacement analysis of large object areas. This paper reports on the use of a digital holographic interferometer to measure displacements over an area of $1.14m2$. The object under study is a framed working table covered with a Formica layer fixed to a granite bench, and it is observed and illuminated employing a high speed and high resolution camera and a continuous wave high output power laser, respectively. A stabilization procedure needs to be established as long illumination distances are required in order to retrieve the entire surface optical phase during a series of continuous deformations. As a proof of principle, two different tests are presented: the first involves a slow continuous loading process and the second a vibration condition. The wrapped phase and displacement maps are both displayed.

Dennis Gabor’s seminal idea of a simple all-optical setup aimed at reconstructing the object wavefront stored on a photographic plate gave birth a little over seven decades ago to the field of holography. In 1971 Gabor obtained the Nobel Prize in Physics for this invention. Still, the road in the early days after his two first papers on the subject was one full of obstacles, so his scientific and engineering contemporaries put his idea to rest for more than 10 years, until the invention of the laser. This fact made his holographic concept take off to new and unsuspected applications. This invited review paper is a homage to Dennis Gabor’s 50th anniversary of his Nobel Prize accolade. For this purpose, the review departs from the typical common route, i.e., those written following a timeline fashion, and instead is written with the intent to cover only a few of the holography applications in optics while scanning the electromagnetic spectrum. In doing this, the authors are aware that other invited papers for this special issue will tackle other subjects not dealt with in this review non-timeline paper.

Recently an optoelectronic holography system was deployed in the clinic with the purpose of quantifying the tympanic membrane (TM) displacements of various mammal species, the objective being the understanding of their middle ear biomechanics. The optoelectronic holography system has an in-line configuration where the data gathered is decoded using lensless digital holography with the Fresnel approximation. This paper presents quantitative data obtained from an acoustically excited postmortem chinchilla’s TM. To achieve this we used a robust customized windowed unwrapping method to unwrap the noisy optical phase obtained by subtracting phase maps of two recorded holograms and the results were compared with those obtained when using the unwrapping branch-cut algorithm. Additionally, phase maps obtained by the phase-stepping technique were compared applying both unwrapping methods. For in vivo applications particular emphasis is made on post-processing dual-shot-acquisition of holograms as one of various acquisition strategies and algorithms to diminish measurement error due to heartbeat, breathing, and patient’s head motion as well as environment induced mechanical disturbances present in a noncontrolled environment, such as in a clinic. By recording only two holograms representing a stationary and deformed state of eardrum, respectively, we can increase the acquisition speed of the camera used to record faster events happening on the TM surface.

Bee honey is an exceptionally nutritious food with unique chemical and mineral contents. This report introduces the use of the second-harmonic generation (SHG) microscopy for imaging honey sugar crystals’ morphology as an alternative for its authentication process. The crystals and their boundaries are clearly observed with SHG compared with bright-field microscopy, where the liquid honey avoids the visualization of a sharp image. Four different honey samples of Mexico’s various floral origins and geographical regions are analyzed in our study. These samples are representative of the diversity and valuable quality of bee honey production. The SHG image information is complemented with Raman spectroscopy (RS) analysis, since this optical technique is widely used to validate the bee’s honey composition stated by its floral origin. We relate the SHG imaging of honey crystals with the well-defined fructose and glucose peaks measured by RS. Size measurement is introduced using the crystal´s length ratio to differentiate its floral origin. From our observations, we can state that SHG is a promising and suitable technique to provide a sort of optical fingerprint based on the floral origin of bee honey.

Second harmonic generation (SHG) microscopy is applied to image honey sugar crystals, with the advantage of avoiding the surrounding signal of the liquid honey. It is a non-contact method where the specialized preparation of the honey is not required. Since the crystallization process of honey depends upon its floral origin, among other conditions, analyzing its crystallographic behavior results in a challenging task as the crystals are mixed and overlapped. This work introduces a simple, multi-variable data analysis requiring only one SHG image, where its gray-level histogram was retrieved and processed from eight Mexican bee honey samples. These honey samples represent a broad range of regions from the central–east part of Mexico, describing different environments and climates. The results obtained with this simplified methodology can differentiate among the botanical origin of the honey samples at different crystallization times and storage conditions. A repeatability test was performed using three honey samples with different crystal morphologies to expand the method to all of the samples. The results show that the proposed methodology could be a helpful alternative to identifying the botanical origin of honey despite its crystallization time.