This editorial reflects on the shape of biomedical engineering as a discipline, and its relation to biomedical optics.

Photoacoustic (PA) imaging is an emerging medical imaging modality that combines optical excitation and ultrasound detection. Because ultrasound scatters much less than light in biological tissues, PA generates high-resolution images at centimeters depth. In recent years, wavelengths in the second near-infrared (NIR-II) window (1000 to 1700 nm) have been increasingly explored due to its potential for preclinical and clinical applications. In contrast to the conventional PA imaging in the visible (400 to 700 nm) and the first NIR-I (700 to 1000 nm) window, PA imaging in the NIR-II window offers numerous advantages, including high spatial resolution, deeper penetration depth, reduced optical absorption, and tissue scattering. Moreover, the second window allows a fivefold higher light excitation energy density compared to the visible window for enhancing the imaging depth significantly. We highlight the importance of the second window for PA imaging and discuss the various NIR-II PA imaging systems and contrast agents with strong absorption in the NIR-II spectral region. Numerous applications of NIR-II PA imaging, including whole-body animal imaging and human imaging, are also discussed.

Elastography imaging is a promising tool—in both research and clinical settings—for diagnosis, staging, and therapeutic treatments of various life-threatening diseases (including brain tumors, breast cancers, prostate cancers, and Alzheimer’s disease). Large variation in the physical (elastic) properties of tissue, from normal to diseased stages, enables highly sensitive characterization of pathophysiological states of the diseases. On the other hand, over the last decade or so, photoacoustic (PA) imaging—an imaging modality that combines the advantageous features of two separate imaging modalities, i.e., high spatial resolution and high contrast obtainable, respectively, from ultrasound- and optical-based modalities—has been emerging and widely studied. Recently, recovery of elastic properties of soft biological tissues—in addition to prior reported recovery of vital tissue physiological information (Hb, HbO₂, SO, and total Hb), noninvasively and nondestructively, with unprecedented spatial resolution (μm) at penetration depth (cm)—has been reported. Studies demonstrating that combined recovery of mechanical tissue properties and physiological information—by a single (PA) imaging unit—pave a promising platform in clinical diagnosis and therapeutic treatments. We offer a comprehensive review of PA imaging technology, focusing on recent advances in relation to elastography. Our review draws out technological challenges pertaining to PA elastography (PAE) imaging, and viable approaches. Currently, PAE imaging is in the nurture stage of its development, where the technology is limited to qualitative study. The prevailing challenges (specifically, quantitative measurements) may be addressed in a similar way by which ultrasound elastography and optical coherence elastography were accredited for quantitative measurements.

Near-infrared spectroscopy (NIRS) is emerging as a rapid, low-cost approach for point-of-care triage of hematomas resulting from traumatic brain injury. However, there remains a lack of standardized test methods for benchtop performance assessment of these devices and incomplete understanding of relevant light–tissue interactions. We propose a phantom-based test method for systems operating near the 800-nm oxy-/deoxy-hemoglobin isosbestic point and implement it to evaluate a clinical system. Semi-idealized phantom geometries are designed to represent epidural/subdural, subarachnoid, and intracerebral hemorrhages. Measurements of these phantoms are made with a commercial NIRS-based hematoma detector to quantify the effect of hematoma type, depth, and size, as well as measurement repeatability and detector positioning relative to the hematoma. Results indicated high sensitivity to epidural/subdural and subarachnoid hematomas. Intracerebral hematomas are detectable to a maximum depth of ∼2.5  cm, depending on thickness and diameter. The maximum lateral detection area for the single-emitter/single-collector device studied here appears elliptical and decreases strongly with inclusion depth. Overall, this study provides unique insights into hematoma detector function and indicates the utility of modular polymer tissue phantoms in performance tests for emerging NIRS-based cerebral diagnostic technology.

We analyze transducer-matched multipulse excitation as a method for improving of the signal-to-noise ratio (SNR) for diode laser-based photoacoustic systems. We discuss the principle of the technique, its advantages, and potential drawbacks and perform measurements to analyze the obtainable SNR increase. We show in experiment and computationally that a lower boundary estimate of 1.2 to 1.8 fold SNR improvement can be provided using transducer-matched pulse bursts, depending on the transducer and particular arrangement. Finally, we analyze implications that the transducer resonance effects may have on the recently introduced advanced photoacoustic techniques. The findings are of immediate interest to modalities utilizing dense pulse sequences and systems possessing limited pulse energy. In particular, transducer-matched multipulse excitation may be beneficial for diode-based photoacoustic systems operated with transducers in the range of 1 to 5 MHz since the required hardware is readily available.

To identify the microstructural modification of the corneal layers during the course of the disease, optical technologies have been pushing the boundary of innovation to achieve cellular resolution of deep layers of the cornea. Gabor-domain optical coherence microscopy (GD-OCM), an optical coherence tomography-based technique that can achieve an isotropic of ∼2-μm resolution over a volume of 1  mm  ×  1  mm  ×  1.2  mm, was developed to investigate the microstructural modifications of corneal layers in four common corneal diseases. Since individual layer visualization without cutting through several layers is challenging due to corneal curvature, a flattening algorithm was developed to remove the global curvature of the endothelial layer and display the full view of the endothelium and Descemet’s membrane in single en face images. As a result, GD-OCM revealed the qualitative changes in size and reflectivity of keratocytes in Fuchs endothelial corneal dystrophy (FECD), which varied by the degree of disease. More importantly, elongated shape and hyperactivation characteristics of keratocytes, associated with the early development of guttae, appeared to start in the posterior stroma very early in the disease process and move toward the anterior stroma during disease progression. This work opens a venue into the pathogenesis of FECD.

The effects of dispersion on optical coherence tomography (OCT) images have long been documented. The imbalance of spectral broadening, caused by dispersion mismatches in the two arms of the OCT interferometer, can result in significant resolution degradation. Efforts to correct this phenomenon have resulted in improved image quality using various techniques. However, dispersion is also present and varies in tissues. As a result, group velocity dispersion (GVD) can be used to detect changes in tissues and provide useful information for diagnosis. Several methods can be utilized to measure the GVD from OCT images: (i) the degradation of the point spread function (PSF), (ii) the shift (walk-off) between images taken at different wavelengths, (iii) the changes in the second derivative of the spectral phase, as well as two new methods, which do not require a reflector and are applicable in intact tissues, i.e., using (iv) the speckle degradation, and (v) the speckle cross correlation. A systematic, experimental, evaluation of these methods is presented to elucidate the capabilities, the limitations, and the accuracy of each technique when attempting to estimate the GVD in scattering samples. The most precise values were obtained from the estimation of the PSF degradation, whereas using the phase derivative method was only applicable to minimally scattering samples. Speckle broadening appears to be the most robust method for tissue GVD measurements.

Given that breast cancer is the second leading cause of cancer-related deaths among women in the United States, it is necessary to continue improving the sensitivity and specificity of breast imaging systems that diagnose breast lesions. Photoacoustic (PA) imaging can provide functional information during in vivo studies and can augment the structural information provided by ultrasound (US) imaging. A full-ring, all-reflective, illumination system for photoacoustic tomography (PAT) coupled to a full-ring US receiver is developed and tested. The US/PA tomography system utilizes a cone mirror and conical reflectors to optimize light delivery for PAT imaging and has the potential to image objects that are placed within the ring US transducer. The conical reflector used in this system distributes the laser energy over a circular cross-sectional area, thereby reducing the overall fluence. This, in turn, allows the operator to increase the laser energy achieving better cross-sectional penetration depth. A proof-of-concept design utilizing a single cone mirror and a parabolic reflector is used for imaging cylindrical phantoms with light-absorbing objects. For the given phantoms, it has been shown that there was no restriction in imaging a given targeted cross-sectional area irrespective of vertical depth, demonstrating the potential of mirror-based, ring-illuminated PAT system. In addition, the all-reflective ring illumination method shows a uniform PA signal across the scanned cross-sectional area.

Optical coherence tomography angiography (OCTA) provides in-vivo images of microvascular perfusion in high resolution. For its application to basic and clinical research, an automatic and robust quantification of the capillary architecture is mandatory. Only this makes it possible to reliably analyze large amounts of image data, to establish biomarkers, and to monitor disease developments. However, due to its optical properties, OCTA images of skin often suffer from a poor signal-to-noise ratio and contain imaging artifacts. Previous work on automatic vessel segmentation in OCTA mostly focuses on retinal and cerebral vasculature. Its applicability to skin and, furthermore, its robustness against imaging artifacts had not been systematically evaluated. We propose a segmentation method that improves the quality of vascular quantification in OCTA images even if corrupted by imaging artifacts. Both the combination of image processing methods and the choice of their parameters are systematically optimized to match the manual labeling of an expert for OCTA images of skin. The efficacy of this optimization-based vessel segmentation is further demonstrated on sample images as well as by a reduced error of derived quantitative vascular network characteristics.

We demonstrate improved optical sectioning in light sheet fluorescence microscopy using tunable structured illumination (SI) frequencies to optimize image quality in scattering specimens. The SI patterns are generated coherently using a one-dimensional spatial light modulator for maximum pattern contrast, and the pattern spatial frequency is adjustable up to half the incoherent cutoff frequency of our detection objective. At this frequency, we demonstrate background reductions of 2 orders of magnitude.

Coherent anti-Stokes Raman scattering (CARS) generates a strong label-free signal in the long wavenumber C─H stretching region. Lipid-rich myelinated tissues, such as brain and spinal cord, would appear to be ideal subjects for imaging with CARS laser-scanning microscopy. However, the highly ordered, biochemically complex, and highly scattering nature of such tissues complicate the use of the technique. A CARS microscopy approach is presented that overcomes the challenges of imaging myelinated tissue to achieve chemically and orientationally sensitive high-resolution images.

We used phase microscopy and Raman spectroscopic measurements to assess the response of in vitro rat C6 glial cells following methamphetamine treatment in real time. Digital holographic microscopy (DHM) and three-dimensional (3-D) tomographic nanoscopy allow measurements of live cell cultures, which yield information about cell volume changes. Tomographic phase imaging provides 3-D information about the refractive index distribution associated with the morphology of biological samples. DHM provides similar information, but for a larger population of cells. Morphological changes in cells are associated with alterations in cell cycle and initiation of cell death mechanisms. Raman spectroscopy measurements provide information about chemical changes within the cells. Our Raman data indicate that the chemical changes in proteins preceded morphological changes, which were seen with DHM. Our study also emphasizes that tomographic phase imaging, DHM, and Raman spectroscopy are imaging tools that can be utilized for noninvasive simultaneous monitoring of morphological and chemical changes in cells during apoptosis and can also be used to monitor other dynamic cell processes.