Viewing life without labels under optical microscopes

Ghosh, Biswajoy; Agarwal, Krishna

doi:10.1038/s42003-023-04934-8

Cited by 22 publications

(9 citation statements)

References 159 publications

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“…A long-sought ideal is to identify early modifications in the matrix density label-free with high-resolution optical microscopy. The occurrence of several autofluorescent molecules in biological cells and tissues 15 – 17 has prompted their use for label-free investigations using conventional optical microscopes in several niche areas like cancer diagnosis 18 and regenerative medicine 19 . Autofluorescence seems to be ideal for super-resolution imaging 8 , 10 as it is source-emitted (better localization and fidelity in distribution); therefore, it overcomes labeling limitations.…”

Section: Introductionmentioning

confidence: 99%

Molecular histopathology of matrix proteins through autofluorescence super-resolution microscopy

Ghosh,

Chatterjee,

Paul

et al. 2024

Sci Rep

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Extracellular matrix diseases like fibrosis are elusive to diagnose early on, to avoid complete loss of organ function or even cancer progression, making early diagnosis crucial. Imaging the matrix densities of proteins like collagen in fixed tissue sections with suitable stains and labels is a standard for diagnosis and staging. However, fine changes in matrix density are difficult to realize by conventional histological staining and microscopy as the matrix fibrils are finer than the resolving capacity of these microscopes. The dyes further blur the outline of the matrix and add a background that bottlenecks high-precision early diagnosis of matrix diseases. Here we demonstrate the multiple signal classification method-MUSICAL-otherwise a computational super-resolution microscopy technique to precisely estimate matrix density in fixed tissue sections using fibril autofluorescence with image stacks acquired on a conventional epifluorescence microscope. We validated the diagnostic and staging performance of the method in extracted collagen fibrils, mouse skin during repair, and pre-cancers in human oral mucosa. The method enables early high-precision label-free diagnosis of matrix-associated fibrotic diseases without needing additional infrastructure or rigorous clinical training.

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Section: Introductionmentioning

confidence: 99%

Molecular histopathology of matrix proteins through autofluorescence super-resolution microscopy

Ghosh,

Chatterjee,

Paul

et al. 2024

Sci Rep

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“…In a short span of less than three decades, several important breakthroughs in light microscopy have revolutionized life sciences. These include, but are not limited to: (i) genetically encoded fluorescent proteins for live cell imaging 4 – 6 , (ii) light sheet microscopy 7 – 12 , (iii) super-resolution microscopy 13 – 24 , (iv) label-free imaging approaches 25 – 29 , (v) machine learning 30 – 34 , and (vi) imaging technologies capable of adapting to the biology of the specimens 35 – 46 . Together, these innovations have allowed biologists to understand the fundamental processes of life across a large range of spatiotemporal scales or under conditions previously considered incompatible with imaging.…”

Section: Introductionmentioning

confidence: 99%

“…Effective checks include assessing whether the labeled samples maintain normal morphology and behavior, and whether consistent results are obtained by using different tags. In certain scenarios, the intrinsic optical characteristics of biological molecules can be leveraged for label-free imaging 26 , 29 . Quantitative phase imaging 132 , 133 , stimulated Raman microscopy 134 – 136 (Fig.…”

Section: Introductionmentioning

confidence: 99%

Imagining the future of optical microscopy: everything, everywhere, all at once

Balasubramanian,

Hobson,

Chew

et al. 2023

Commun Biol

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The optical microscope has revolutionized biology since at least the 17th Century. Since then, it has progressed from a largely observational tool to a powerful bioanalytical platform. However, realizing its full potential to study live specimens is hindered by a daunting array of technical challenges. Here, we delve into the current state of live imaging to explore the barriers that must be overcome and the possibilities that lie ahead. We venture to envision a future where we can visualize and study everything, everywhere, all at once – from the intricate inner workings of a single cell to the dynamic interplay across entire organisms, and a world where scientists could access the necessary microscopy technologies anywhere.

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“…Traditional optical observation methods encounter challenges such as limited depth of penetration and difficulties in imaging live samples 20 . Additionally, the use of lasers in conventional methods poses a potential risk of damaging sensitive biological samples 21 .…”

Section: Introductionmentioning

confidence: 99%

Uncertainty Quantification in Acoustic Impedance of Atlantic Salmon Fish Scale using Scanning Acoustic Microscopy

Agarwal,

Ojha,

Dalmo

et al. 2023

Preprint

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Scanning Acoustic Microscopy (SAM) emerges as a versatile label-free imaging technology with broad applications in biomedical imaging, non-destructive testing, and material research. This article presents a framework for the estimation of stochastic impedance through SAM, with a particular focus on its application to the salmon fish scale. The framework leverages uncertain reflectance, marking its pioneering application to uncertainty quantification in the acoustic impedance of fish scales through acoustic responses. The study uses maximal overlap discrete wavelet transform, to decompose acoustic responses effectively and is further processed to predict the acoustic impedance. To establish the effectiveness of the proposed framework, well-known materials like a pair of target medium (polyvinylidene fluoride) and reference medium (polyimide) are employed for impedance characterization. Results demonstrate over 90%accuracy in PVDF impedance estimation, validating the framework. A stochastic impedance map, using Kriging with a Gaussian variogram, offers insights into the complex biomechanics of a fish’s scale.

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Viewing life without labels under optical microscopes

Cited by 22 publications

References 159 publications

Molecular histopathology of matrix proteins through autofluorescence super-resolution microscopy

Molecular histopathology of matrix proteins through autofluorescence super-resolution microscopy

Imagining the future of optical microscopy: everything, everywhere, all at once

Uncertainty Quantification in Acoustic Impedance of Atlantic Salmon Fish Scale using Scanning Acoustic Microscopy

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