Rapid and label-free histological imaging of unprocessed surgical tissues via dark-field reflectance ultraviolet microscopy

Ye, Shiwei; Zou, Junjie; Huang, Chenming; Feng, Xiang; Wen, Zong-Hua; Wang, Nannan; Yu, Jia; He, Yu; Liu, Peng; Xin, Mei; Li, Hui; Niu, Lili; Gong, Peng; Zheng, Wei

doi:10.1016/j.isci.2022.105849

Cited by 8 publications

(3 citation statements)

References 53 publications

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“…Recently, it has been demonstrated for fast hematology analysis of whole blood smears and blood samples in custom microfluidic cartridges for point-of-care applications enabled by advancements in low-cost UV LEDs and sensors [9][10][11][12][13]. UV microscopy has also been used for multi-spectral, label-free histopathology of tissue samples [14][15][16]. In these prior works, deep neural networks have been required to pseudo-colorize images to mimic conventional biochemical stains (i.e., hematoxylin and eosin, Giemsa) [12,13,15].…”

Section: Introductionmentioning

confidence: 99%

Deep-ultraviolet microscopy for analysis of bone marrow aspirate adequacy

Gorti,

Subramanian,

Aljudi

et al. 2024

Optical Diagnostics and Sensing XXIV: Toward Point-of-Care Diagnostics

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Bone marrow aspiration procedures play an important role in the assessment of patients with blood and marrow diseases, including cancers. Evaluating the adequacy of aspirates, indicated by the presence of bony spicules, is crucial to ensure the procedural success and collection of relevant diagnostic material. Unfortunately, inadequate samples occur in approximately 50 % of cases, requiring patients to undergo repeat procedures. This is particularly problematic for pediatric patients who need to be anesthetized before each procedure. The current gold standard is hematopathologist examination of Giemsa-stained slides, which is time consuming and requires expensive biochemical reagents and trained technicians. Recently, Here we present a portable, LED-based UV microscope designed for real-time inspection of bone marrow aspirates. We discuss results from a clinical trial with pediatric oncology patients demonstrating excellent agreement between UV examination of unstained slides and ground truth pathologist examination of stained slides. Furthermore, we demonstrate whole slide imaging using a previously developed, compact UV microscopy system and automated spicule detection with deep neural networks to work towards point-of-care applications.

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Section: Introductionmentioning

confidence: 99%

Deep-ultraviolet microscopy for analysis of bone marrow aspirate adequacy

Gorti,

Subramanian,

Aljudi

et al. 2024

Optical Diagnostics and Sensing XXIV: Toward Point-of-Care Diagnostics

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“…To this end, different widefield label-free imaging techniques have recently been developed for slide-free histological imaging. For instance, dark-field reflectance ultraviolet microscopy (DRUM) [ 9 ] and computational high-throughput autofluorescence microscopy by pattern illumination (CHAMP) [ 10 ] are two examples that have demonstrated the use of the high absorption property of deep-UV in cell nuclei to provide nuclear contrast on fresh and unprocessed tissues. DRUM is a simple light-emitting diode (LED)-based widefield histological imaging technique that leverages both dark field reflectance contrast for hematoxylin analog and single emission channel autofluorescence contrast for eosin analog.…”

Section: Introductionmentioning

confidence: 99%

Deep learning-assisted low-cost autofluorescence microscopy for rapid slide-free imaging with virtual histological staining

Wong,

Chen,

Shi

et al. 2024

Biomed. Opt. Express

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Slide-free imaging techniques have shown great promise in improving the histological workflow. For example, computational high-throughput autofluorescence microscopy by pattern illumination (CHAMP) has achieved high resolution with a long depth of field, which, however, requires a costly ultraviolet laser. Here, simply using a low-cost light-emitting diode (LED), we propose a deep learning-assisted framework of enhanced widefield microscopy, termed EW-LED, to generate results similar to CHAMP (the learning target). Comparing EW-LED and CHAMP, EW-LED reduces the cost by 85×, shortening the image acquisition time and computation time by 36× and 17×, respectively. This framework can be applied to other imaging modalities, enhancing widefield images for better virtual histology.

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“…Traditional methods such as immunochemistry and histology rely on extensive fixation or cryo-freezing protocols, followed by laborious multi-step staining procedures. All of these processes can destroy scaffolds and alter the presentation of deposited ECM in both research [38] and clinical settings [39, 40]. Molecular techniques, such as qPCR, offer insight on gene expression that might not reflect protein deposition nor spatial distribution.…”

Section: Introductionmentioning

confidence: 99%

Extensive collagen deposition by mesenchymal stem cells cultured in 3D self-assembled peptide scaffolds as revealed by nanoplasmonic colorimetric histology

Chong

Charnley

Ratcliffe

et al. 2022

Preprint

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Ultrashort self-assembling peptides of typically less than 5 amino acids in length can be modified with highly aromatic protecting groups such as Fluorenylmethyloxycarbonyl (Fmoc) resulting in the rapid formation of nanofibrillar hydrogels. These hydrogels show great promise as scaffolds for tissue engineering and 3D cell culture. If further modified with bioactive peptide sequences, the hydrogels can be used to promote and control cell adhesion, and even support the ability for cultured mesenchymal stem cells (MSCs) to differentiate into therapeutically useful lineages, such as chondrocytes. Whilst there have been numerous examples of how the addition of such sequences can promote the bioactivity of cultured cells, the effect adding these sequences has on the nanoarchitecture, mechanical properties and stability of the resultant hydrogels is less well explored. Here, we have performed a series of experiments investigating the effect incorporating stoichiometric amounts of Fmoc-Arg-Gly-Asp (Fmoc-RGD) into Fmoc-Phe-Phe (Fmoc-FF) hydrogels has on nanofibril morphology, and the rheological properties and stability of the formed gels. Furthermore, we correlate these variations in nano-architectural and mechanical properties with their ability to support the culture of clinically valuable bone marrow derived human mesenchymal stem cells (hMSCs). Our results show that at relatively low concentrations Fmoc-RGD is incorporated into the self-assembled Fmoc-FF nanofibrils creating biocompatible hydrogels capable of supporting hMSC growth for up to a week. At increased concentrations of Fmoc-RGD, incorporation into the nanofibrils is lost and the hydrogels that form are no longer stable enough to support the culture of hMSCs for multiple days. These findings shed light on the importance of balancing the positive effects of adding bioactive ligands to 3D scaffolds with the negative effects these ligands can have on the stability and mechanical properties of these scaffolds. Further, the findings will have implications for the choice of ultrashort peptide scaffolds for tissue engineering applications to support MSCs towards chondrogenic differentiation, and their application as gel-cell constructs in cartilage tissue engineering for osteoarthritis.

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Rapid and label-free histological imaging of unprocessed surgical tissues via dark-field reflectance ultraviolet microscopy

Cited by 8 publications

References 53 publications

Deep-ultraviolet microscopy for analysis of bone marrow aspirate adequacy

Deep-ultraviolet microscopy for analysis of bone marrow aspirate adequacy

Deep learning-assisted low-cost autofluorescence microscopy for rapid slide-free imaging with virtual histological staining

Extensive collagen deposition by mesenchymal stem cells cultured in 3D self-assembled peptide scaffolds as revealed by nanoplasmonic colorimetric histology

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