Accurate reconstruction methods are needed to extend the reach of quantitative 3D microscopy to diverse samples in biology with illumination in an arbitrary states of spatial and temporal coherence. Recent research in optical diffraction tomography (ODT) reconstruction has incorporated non-invertible physics-based forward models, such as beam propagation or the Lippman-Schwinger equation, to overcome these effects. However, partially coherent methods have yet to incorporate more accurate physical models, as these artifacts are reduced by the incoherence of the illumination and require heavy computation. Here we leverage the sparsity of the 3D Green’s function solution to the Helmholtz equation in k-space to reduce the problem’s dimensionality to rapidly compute a partially coherent forward model, allowing for a gradient descent-type iterative solver to reconstruct 3D thick biological samples in a variety of illuminations, including broadband light.
We demonstrate a fiber-based quantitative phase imaging (QPI) system with epi-illumination to acquire tissue and cellar level structure. Our approach is based on quantitative oblique back-illumination microscopy (qOBM), which keeps the advantages of QPI—label-free and non-destructive with nanometer-scale sensitivity—while also delivering tomographic sectioning capabilities in thick scattering samples using epi-illumination. The developed system uses a simple and robust configuration consisting of a flexible fiber bundle and a GRIN lens. Here data are presented with histopathological feature analysis. This technique, with its compact setup and real-time processing algorithm, can lead to in-vivo medical diagnosis, for clinical surgery and endoscopy.
The first-line treatment for brain cancer is surgery, which focuses on maximizing the percentage of the tumor removed during surgery (i.e., extent of resection) while minimizing damage to healthy brain tissue. Data show that extent of resection is one of the most critical factors associated with prolonged survival. However, differentiating between tumor and healthy tissue intraoperatively remains a significant clinical challenge, resulting in an exceedingly low 5-year survival rate of only ~35%. In this work, we show that quantitative oblique back illumination microscopy (qOBM), a novel label-free optical imaging technique that achieves tomographic quantitative phase imaging (QPI) in thick scattering samples, clearly differentiates between tumor and healthy tissue. Using a 9L gliosarcoma rat tumor model, we show that quantitative image features from qOBM provide a robust set of biomarkers for disease. In addition, tumor regions, including diffuse tumor, and healthy brain structures, show excellent structural agreement with H&E stained and sliced brightfield images, the gold standard for cancer detection. The unique attribute of qOBM—low-cost, easy-to-use, label-free, and real-time—make this technology ideally suited to help guide neurosurgery and address this important unmet need. Here we describe our free-space qOBM system and present quantitative results from the 9L gliosarcoma rat tumor model.
We have recently developed quantitative oblique back-illumination microscopy (qOBM), which enables full-field quantitative phase imaging (QPI) of objects embedded in a thick scattering medium. This epi-mode technique makes use of multiple scattering as a source of transmissive illumination from within, allowing for rich structural detail based on optical path delay. We now produce quantitative 3D renderings of index of refraction with sub- cellular detail by computing a 3D transfer function of the entire optical system, including the multiply scattered illumination, and deconvolving it from a vertical stack of phase gradient contrast images. This approach allows truly non-invasive, label-free, tomographic quantitative reconstructions if index of refraction in thick scattering samples including thick tissue samples, at low cost and with simple operation, bringing QPI’s unparalleled access to sub-cellular structural detail to previously unavailable domains of investigation.
Quantitative phase imaging (QPI) provides unique access to cellular and subcellular structures with nanometer-scale sensitivity, making it a valuable tool for non-destructive, label-free imaging of biological samples. However, implementation of QPI typically involves a transmission-based geometry and requires thin samples, preventing use of QPI in many important clinical settings, including endoscopy. In this work we demonstrate a fiber-optic device, with epi-illumination, capable of providing quantitative phase information that is well suited for clinical endoscopy, among other biomedical applications.
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