4D imaging and analysis of multicellular tumour spheroid cell migration and invasion

Richards, R. C.; Mason, David; Kelly, Ryan R.; Lévy, Raphaël; Bearon, Rachel N.

doi:10.1101/443648

Cited by 13 publications

(15 citation statements)

References 47 publications

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“…We harnessed this property for wavelength-encoded tagging and tracking of thousands of densely populated cells in a 3D scattering tissue. The spectral encoding offers a compelling advantage for cell identification and tracking, compared to image-based tracking, for example, by light-sheet microscopy, which requires frequent imaging (every few minutes) 41 . Improvements of multiplexing capability are possible.…”

Section: Discussionmentioning

confidence: 99%

Wavelength-encoded laser particles for massively multiplexed cell tagging

et al. 2019

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Large-scale single-cell analyses have become increasingly important given the role of cellular heterogeneity in complex biological systems. However, no current techniques enable optical imaging of uniquely-tagged individual cells. Fluorescence-based approaches can only distinguish a small number of distinct cells or cell groups at a time because of spectral crosstalk between conventional fluorophores. Here we investigate large-scale cell tracking using intracellular laser particles as imaging probes that emit coherent laser light with a characteristic wavelength. Made of silica-coated semiconductor microcavities, these laser particles have single-mode emission over a broad range from 1170 to 1580 nm with sub-nm linewidths, enabling massive spectral Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:

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Section: Discussionmentioning

confidence: 99%

Wavelength-encoded laser particles for massively multiplexed cell tagging

et al. 2019

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“…Interestingly, the skewness is more pronounced for the Monotonic decrease of pressure from the core to periphery of the MCS: To assess the effectiveness of the TPs as sensors of the local microenvironment, we calculated the radial pressure (Eq. (10) in the methods section) profile for both CCs and TPs at t = 7.5τ . In order to distinguish between the local pressure due to the TPs and CCs, we calculated pressure experienced just by the TPs.…”

Section: Simulationsmentioning

confidence: 99%

Tracer particles sense local stresses in an evolving multicellular spheroid without affecting the anomalous dynamics of the cancer cells

Samanta

Sinha

Thirumalai

2022

Preprint

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Measurements of local stresses on the cancer cells (CCs), inferred by embedding inert compressible tracer particles (TPs) in a growing multicellular spheroid (MCS), show that pressure decreases monotonically as the distance from the core of the MCS increases. How faithfully do the TPs report the local stresses in the CCs is an important question because pressure buildup in the MCS is dynamically generated due to CC division, which implies that the CC dynamics should be minimally altered by the TPs. Here using theory and simulations, we show that although the TP dynamics is unusual, exhibiting sub-diffusive behavior on times less than the CC division times and hyper-diffusive dynamics on in the long-time limit, they do not affect the long-time CC dynamics or the local CC stress distributions. The CC pressure profile within the MCS, which decays from a high value at the core to the periphery, is almost identical with and without the TPs. That the TPs have insignificant effect on the local stresses in the MCS implies that they are reliable reporters of the CC microenvironment.

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“…The ability of fluorescent proteins to be genetically encoded into a cell as a non-invasive fluorescent tag, without the need for fixing or permeabilising cells, makes them indispensable for live cell imaging, and may be increasingly useful in 4D imaging of opaque samples such as spheroids [ 202 ], where the penetration depth of antibodies and dyes may be limited. Imaging in four dimensions can shed light on intracellular dynamics, such as protein trafficking and organelle interplay [ 203 ], and has also been applied to small single-celled systems such as yeast [ 204 ].…”

Section: Imaging Agents Used For Fluorescence Microscopymentioning

confidence: 99%

Fluorescence Microscopy—An Outline of Hardware, Biological Handling, and Fluorophore Considerations

Hickey

Ung

Bader

et al. 2021

Cells

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Fluorescence microscopy has become a critical tool for researchers to understand biological processes at the cellular level. Micrographs from fixed and live-cell imaging procedures feature in a plethora of scientific articles for the field of cell biology, but the complexities of fluorescence microscopy as an imaging tool can sometimes be overlooked or misunderstood. This review seeks to cover the three fundamental considerations when designing fluorescence microscopy experiments: (1) hardware availability; (2) amenability of biological models to fluorescence microscopy; and (3) suitability of imaging agents for intended applications. This review will help equip the reader to make judicious decisions when designing fluorescence microscopy experiments that deliver high-resolution and informative images for cell biology.

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4D imaging and analysis of multicellular tumour spheroid cell migration and invasion

Cited by 13 publications

References 47 publications

Wavelength-encoded laser particles for massively multiplexed cell tagging

Wavelength-encoded laser particles for massively multiplexed cell tagging

Tracer particles sense local stresses in an evolving multicellular spheroid without affecting the anomalous dynamics of the cancer cells

Fluorescence Microscopy—An Outline of Hardware, Biological Handling, and Fluorophore Considerations

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