Significance: Tumor heterogeneity poses a challenge for the chemotherapeutic treatment of cancer. Tissue dynamics spectroscopy captures dynamic contrast and can capture the response of living tissue to applied therapeutics, but the current analysis averages over the complicated spatial response of living biopsy samples.
Aim: To develop tissue dynamics spectroscopic imaging (TDSI) to map the heterogeneous spatial response of tumor tissue to anticancer drugs.
Approach: TDSI is applied to tumor spheroids grown from cell lines and to ex vivo living esophageal biopsy samples. Doppler fluctuation spectroscopy is performed on a voxel basis to extract spatial maps of biodynamic biomarkers. Functional images and bivariate spatial maps are produced using a bivariate color merge to represent the spatial distribution of pairs of signed drug-response biodynamic biomarkers.
Results: We have mapped the spatial variability of drug responses within biopsies and have tracked sample-to-sample variability. Sample heterogeneity observed in the biodynamic maps is associated with histological heterogeneity observed using inverted selective-plane illumination microscopy.
Conclusion: We have demonstrated the utility of TDSI as a functional imaging method to measure tumor heterogeneity and its potential for use in drug-response profiling.
Diagnoses performed on the basis of histopathological evaluation depend on the premise that information derived from a small number of samples is valid for the entire tissue volume. By insufficiently sampling a biopsy volume the ability of pathologists to draw meaningful inferences from the sample is impeded. This work attempts to apply an information theoretic approach to biopsy sampling rates informed by variation in tissue morphology identified by persistent homology. By quantifying the diagnostic information present in a sample may be possible to prevent under sampling by the clinician by creating a “Nyquist limit" for histopathological sampling given the frequency of morphologically distinct regions in a single biopsy.
The use of iSPIM and optical-clearing enables 3D high-content histological imaging of large tissues with subcellular resolution and almost 2 mm imaging depth, providing 3D structural information for generating tissue models or for validating low-resolution imaging modalities.
Prostate cancer comprises the second most common cancer in men. One of the most powerful and established prognostic indicators of adenocarcinoma of the prostate is the Gleason score, a subjective assessment of the pattern of tumor growth and extent of glandular differentiation in H&E stained histology slides. Despite being the most dominant prostate grading method in use, the Gleason score suffers from high variability between grading pathologists, and due to its 2D nature, fails to effectively capture potentially prognostic information contained in 3D glandular growth patterns. We have previously demonstrated that persistent homology, a subset of topological data analysis (TDA), is effective in generating a quantitative morphological descriptor capable of differentiating Gleason grade in 2D. By capturing glands as loops in 2D, and voids in 3D, persistent homology lends itself naturally to the assessment of 3D glandular growth patterns while maintaining a correspondence to their 2D analogue. Dual-view inverted selective plane illumination microscopy (diSPIM) with a fluorescent H&E analogue was leveraged for volumetric imaging of optically-cleared prostate biopsies. The two orthogonal views of the diSPIM system yielded isotropic resolution in all dimensions, facilitating reconstruction of tissue histology in 3D for quantitative morphological assessment by persistent homology. The use of a nuclei specific hematoxylin analog (DRAQ5), in addition to the isotropic resolution of the system, enabled accurate 3D nuclei segmentation, thereby facilitating application of persistent homology to the corresponding nuclei 3D point clouds. Through TDA a quantitative, reproducible descriptor for 3D prostate cancer morphology will be demonstrated.
Current gold-standard histopathology for cancerous biopsies is destructive, time consuming, and limited to 2D slices, which do not faithfully represent true 3D tumor micro-morphology. Light sheet microscopy has emerged as a powerful tool for 3D imaging of cancer biospecimens. Here, we utilize the versatile dual-view inverted selective plane illumination microscopy (diSPIM) to render digital histological images of cancer biopsies. Dual-view architecture enabled more isotropic resolution in X, Y, and Z; and different imaging modes, such as adding electronic confocal slit detection (eCSD) or structured illumination (SI), can be used to improve degraded image quality caused by background signal of large, scattering samples. To obtain traditional H&E-like images, we used DRAQ5 and eosin (D&E) staining, with 488nm and 647nm laser illumination, and multi-band filter sets. Here, phantom beads and a D&E stained buccal cell sample have been used to verify our dual-view method. We also show that via dual view imaging and deconvolution, more isotropic resolution has been achieved for optical cleared human prostate sample, providing more accurate quantitation of 3D tumor architecture than was possible with single-view SPIM methods. We demonstrate that the optimized diSPIM delivers more precise analysis of 3D cancer microarchitecture in human prostate biopsy than simpler light sheet microscopy arrangements.
Current methods for breast tumor margin detection are invasive, time consuming, and typically result in a reoperative rate of over 25%. This marks a clear clinical need to develop improved tools to intraoperatively differentiate negative versus positive tumor margins. Here, we utilize photoacoustic tomography (PAT), ultrasound (US), and inverted Selective Plane Illumination Microscopy (iSPIM) to assess breast tumor margins in eight human breast biopsies. Our PAT/US system consists of a tunable Nd:YAG laser (NT 300, EKSPLA) coupled with a 40MHz central frequency US probe (Vevo2100, FUJIFILM Visual Sonics). This system allows for the delivery of 10Hz, 5ns pulses with fluence of 40mJ/cm2 to the tissue with PAT and US axial resolutions of 125μm and 40μm, respectively. For this study, we used a linear stepper motor to acquire volumetric PAT/US images of the breast biopsies using 1100nm light to identify bloodrich “tumor” regions and 1210nm light to identify lipid-rich “healthy” regions. iSPIM (Applied Scientific Instrumentation) is an advanced microscopy technique with lateral resolution of 1.5μm and axial resolution of 7μm. We used 488nm laser excitation and acridine orange as a general comprehensive histology stain. Our results show that PAT/US can be used to identify lipid-rich regions, dense areas of arterioles and arteries, and other internal structures such as ducts. iSPIM images correlate well with histopathology slides and can verify nuclear features, cell type and density, stromal features, and microcalcifications. Together, this multimodality approach has the potential to improve tumor margin detection with a high degree of sensitivity and specificity.
High-throughput optical imaging is critical to obtain large-scale neural connectivity information of brain in neuroscience. Using a digital mirror device and a scientific complementary metal-oxide semiconductor camera, we report a significant speed improvement of structured illumination microscopy (SIM), which produces a maximum SIM net frame rate of 133 Hz. We perform three-dimensional (3-D) imaging of mouse brain slices at diffraction-limited resolution and demonstrate the fast 3-D imaging capability to a large sample with an imaging rate of 6.9×10 7 pixel/s of our system, an order of magnitude faster than previously reported.
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.