@article {69578, title = {A high-throughput microfabricated platform for rapid quantification of metastatic potential.}, journal = {Sci Adv}, volume = {10}, year = {2024}, month = {2024 Aug 16}, pages = {eadk0015}, abstract = {

Assays that measure morphology, proliferation, motility, deformability, and migration are used to study the invasiveness of cancer cells. However, native invasive potential of cells may be hidden from these contextual metrics because they depend on culture conditions. We created a micropatterned chip that mimics the native environmental conditions, quantifies the invasive potential of tumor cells, and improves our understanding of the malignancy signatures. Unlike conventional assays, which rely on indirect measurements of metastatic potential, our method uses three-dimensional microchannels to measure the basal native invasiveness without chemoattractants or microfluidics. No change in cell death or proliferation is observed on our chips. Using six cancer cell lines, we show that our system is more sensitive than other motility-based assays, measures of nuclear deformability, or cell morphometrics. In addition to quantifying metastatic potential, our platform can distinguish between motility and invasiveness, help study molecular mechanisms of invasion, and screen for targeted therapeutics.

}, keywords = {Cell Line, Tumor, Cell Movement, Cell Proliferation, High-Throughput Screening Assays, Humans, Lab-On-A-Chip Devices, Microtechnology, Neoplasm Invasiveness, Neoplasm Metastasis, Neoplasms}, issn = {2375-2548}, doi = {10.1126/sciadv.adk0015}, url = {https://doi.org/10.1126/sciadv.adk0015}, author = {Bhattacharya, Smiti and Ettela, Abora and Haydak, Jonathan and Hobson, Chad M and Stern, Alan and Yoo, Miran and Chew, Teng-Leong and Gusella, G Luca and Gallagher, Emily J and Hone, James C and Azeloglu, Evren U} } @article {69219, title = {More than just {\textquoteright}added value{\textquoteright}: The perils of not establishing shared core facilities in resource-constrained communities.}, journal = {Journal of Microscopy}, year = {2024}, month = {2024 Feb 20}, abstract = {

The accelerating pace of technological advancements necessitates specialised expertise and cutting-edge instruments to maintain competitive research in life sciences. Core facilities - collaborative laboratories equipped with state-of-the-art tools and staffed by expert personnel - are vital resources that support diverse scientific endeavours. However, their adoption in lower-income communities has been comparatively stagnant due to both financial and cultural challenges. This paper explores the perils of not supporting core facilities on national research enterprises, underscoring the need for balanced investments in discovery science and crucial infrastructure support. We explore the implications from the perspectives of funders, university leaders and lab heads. We advocate for a paradigm shift to recognise these facilities as essential components of national research efforts. Core facilities are positioned not as optional but as strategic investments that can catalyse breakthroughs, particularly in environments with limited resources.

}, issn = {1365-2818}, doi = {10.1111/jmi.13277}, author = {Rahmoon, Mai A and Hobson, Chad M and Aaron, Jesse S and Balasubramanian, Harikrushnan and Chew, Teng-Leong} } @article {69321, title = {More than just {\textquoteright}added value{\textquoteright}: The perils of not establishing shared core facilities in resource-constrained communities.}, journal = {J Microsc}, year = {2024}, month = {2024 Feb 20}, abstract = {

The accelerating pace of technological advancements necessitates specialised expertise and cutting-edge instruments to maintain competitive research in life sciences. Core facilities - collaborative laboratories equipped with state-of-the-art tools and staffed by expert personnel - are vital resources that support diverse scientific endeavours. However, their adoption in lower-income communities has been comparatively stagnant due to both financial and cultural challenges. This paper explores the perils of not supporting core facilities on national research enterprises, underscoring the need for balanced investments in discovery science and crucial infrastructure support. We explore the implications from the perspectives of funders, university leaders and lab heads. We advocate for a paradigm shift to recognise these facilities as essential components of national research efforts. Core facilities are positioned not as optional but as strategic investments that can catalyse breakthroughs, particularly in environments with limited resources.

}, issn = {1365-2818}, doi = {10.1111/jmi.13277}, author = {Rahmoon, Mai A and Hobson, Chad M and Aaron, Jesse S and Balasubramanian, Harikrushnan and Chew, Teng-Leong} } @article {68627, title = {The challenges and opportunities of open-access microscopy facilities.}, journal = {Journal of Microscopy}, year = {2023}, month = {2023 Feb 13}, abstract = {

Microscopy core facilities are increasingly utilized research resources, but they are generally only available to users within the host institution. Such localized access misses an opportunity to facilitate research across a broader user base. Here, we present the model of an open-access microscopy facility, using the Advanced Imaging Center (AIC) at Howard Hughes Medical Institute Janelia Research Campus as an example. The AIC has pioneered a model whereby advanced microscopy technologies and expertise are made accessible to researchers on a global scale. We detail our experiences in addressing the considerable challenges associated with this model for those who may be interested in launching an open-access imaging facility. Importantly, we focus on how this model can empower researchers, particularly those from resource-constrained settings. This article is protected by copyright. All rights reserved.

}, issn = {1365-2818}, doi = {10.1111/jmi.13176}, author = {Cartwright, Heather N and Hobson, Chad M and Chew, Teng-Leong and Reiche, Michael A and Aaron, Jesse S} } @article {68715, title = {Extracellular matrix assembly stress initiates Drosophila central nervous system morphogenesis.}, journal = {Developmental Cell}, year = {2023}, month = {2023 Apr 17}, abstract = {

Forces controlling tissue morphogenesis are attributed to cellular-driven activities, and any role for extracellular matrix (ECM) is assumed to be passive. However, all polymer networks, including ECM, can develop autonomous stresses during their assembly. Here, we examine the morphogenetic function of an ECM before reaching homeostatic equilibrium by analyzing de novo ECM assembly during Drosophila ventral nerve cord (VNC) condensation. Asymmetric VNC shortening and a rapid decrease in surface area correlate with the exponential assembly of collagen IV (Col4) surrounding the tissue. Concomitantly, a transient developmentally induced Col4 gradient leads to coherent long-range flow of ECM, which equilibrates the Col4 network. Finite element analysis and perturbation of Col4 network formation through the generation of dominant Col4 mutations that affect assembly reveal that VNC morphodynamics is partially driven by a sudden increase in ECM-driven surface tension. These data suggest that ECM assembly stress and associated network instabilities can actively participate in tissue morphogenesis.

}, issn = {1878-1551}, doi = {10.1016/j.devcel.2023.03.019}, author = {Serna-Morales, Eduardo and S{\'a}nchez-S{\'a}nchez, Besaiz J and Marcotti, Stefania and Nichols, Angus and Bhargava, Anushka and Dragu, Anca and Hirvonen, Liisa M and Diaz-de-la-Loza, Maria-Del-Carmen and Mink, Matyas and Cox, Susan and Rayfield, Emily and Lee, Rachel M and Hobson, Chad M and Chew, Teng-Leong and Stramer, Brian M} } @article {68784, title = {Extracellular matrix assembly stress initiates Drosophila central nervous system morphogenesis.}, journal = {Developmental Cell}, volume = {58}, year = {2023}, month = {2023 May 22}, pages = {825-835.e6}, abstract = {

Forces controlling tissue morphogenesis are attributed to cellular-driven activities, and any role for extracellular matrix (ECM) is assumed to be passive. However, all polymer networks, including ECM, can develop autonomous stresses during their assembly. Here, we examine the morphogenetic function of an ECM before reaching homeostatic equilibrium by analyzing de novo ECM assembly during Drosophila ventral nerve cord (VNC) condensation. Asymmetric VNC shortening and a rapid decrease in surface area correlate with the exponential assembly of collagen IV (Col4) surrounding the tissue. Concomitantly, a transient developmentally induced Col4 gradient leads to coherent long-range flow of ECM, which equilibrates the Col4 network. Finite element analysis and perturbation of Col4 network formation through the generation of dominant Col4 mutations that affect assembly reveal that VNC morphodynamics is partially driven by a sudden increase in ECM-driven surface tension. These data suggest that ECM assembly stress and associated network instabilities can actively participate in tissue morphogenesis.

}, keywords = {Animals, Central Nervous System, Drosophila, Extracellular Matrix, Morphogenesis}, issn = {1878-1551}, doi = {10.1016/j.devcel.2023.03.019}, author = {Serna-Morales, Eduardo and S{\'a}nchez-S{\'a}nchez, Besaiz J and Marcotti, Stefania and Nichols, Angus and Bhargava, Anushka and Dragu, Anca and Hirvonen, Liisa M and Diaz-de-la-Loza, Maria-Del-Carmen and Mink, Matyas and Cox, Susan and Rayfield, Emily and Lee, Rachel M and Hobson, Chad M and Chew, Teng-Leong and Stramer, Brian M} } @article {69055, title = {Imagining the future of optical microscopy: everything, everywhere, all at once.}, journal = {Communications Biology}, volume = {6}, year = {2023}, month = {2023 Oct 28}, pages = {1096}, abstract = {

The optical microscope has revolutionized biology since at least the 17 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.

}, keywords = {Microscopy}, issn = {2399-3642}, doi = {10.1038/s42003-023-05468-9}, author = {Balasubramanian, Harikrushnan and Hobson, Chad M and Chew, Teng-Leong and Aaron, Jesse S} } @article {68080, title = {Combining multiple fluorescence imaging techniques in biology: when one microscope is not enough.}, journal = {Molecular Biology of the Cell}, volume = {33}, year = {2022}, month = {2022 May 15}, pages = {tp1}, abstract = {

While fluorescence microscopy has proven to be an exceedingly useful tool in bioscience, it is difficult to offer simultaneous high resolution, fast speed, large volume, and good biocompatibility in a single imaging technique. Thus, when determining the image data required to quantitatively test a complex biological hypothesis, it often becomes evident that multiple imaging techniques are necessary. Recent years have seen an explosion in development of novel fluorescence microscopy techniques, each of which features a unique suite of capabilities. In this Technical Perspective, we highlight recent studies to illustrate the benefits, and often the necessity, of combining multiple fluorescence microscopy modalities. We provide guidance in choosing optimal technique combinations to effectively address a biological question. Ultimately, we aim to promote a more well-rounded approach in designing fluorescence microscopy experiments, leading to more robust quantitative insight.

}, keywords = {Biology, Microscopy, Fluorescence, Optical Imaging}, issn = {1939-4586}, doi = {10.1091/mbc.E21-10-0506}, author = {Hobson, Chad M and Aaron, Jesse S} } @article {68409, title = {Practical considerations for quantitative light sheet fluorescence microscopy.}, journal = {Nature Methods}, volume = {19}, year = {2022}, month = {2022 Dec 01}, pages = {1538-49}, abstract = {

Fluorescence microscopy has evolved from a purely observational tool to a platform for quantitative, hypothesis-driven research. As such, the demand for faster and less phototoxic imaging modalities has spurred a rapid growth in light sheet fluorescence microscopy (LSFM). By restricting the excitation to a thin plane, LSFM reduces the overall light dose to a specimen while simultaneously improving image contrast. However, the defining characteristics of light sheet microscopes subsequently warrant unique considerations in their use for quantitative experiments. In this Perspective, we outline many of the pitfalls in LSFM that can compromise analysis and confound interpretation. Moreover, we offer guidance in addressing these caveats when possible. In doing so, we hope to provide a useful resource for life scientists seeking to adopt LSFM to quantitatively address complex biological hypotheses.

}, issn = {1548-7105}, doi = {10.1038/s41592-022-01632-x}, author = {Hobson, Chad M and Guo, Min and Vishwasrao, Harshad D and Wu, Yicong and Shroff, Hari and Chew, Teng-Leong} } @article {67987, title = {When light meets biology - how the specimen affects quantitative microscopy.}, journal = {Journal of Cell Science}, volume = {135}, year = {2022}, month = {2022 Mar 15}, abstract = {

Fluorescence microscopy images should not be treated as perfect representations of biology. Many factors within the biospecimen itself can drastically affect quantitative microscopy data. Whereas some sample-specific considerations, such as photobleaching and autofluorescence, are more commonly discussed, a holistic discussion of sample-related issues (which includes less-routine topics such as quenching, scattering and biological anisotropy) is required to appropriately guide life scientists through the subtleties inherent to bioimaging. Here, we consider how the interplay between light and a sample can cause common experimental pitfalls and unanticipated errors when drawing biological conclusions. Although some of these discrepancies can be minimized or controlled for, others require more pragmatic considerations when interpreting image data. Ultimately, the power lies in the hands of the experimenter. The goal of this Review is therefore to survey how biological samples can skew quantification and interpretation of microscopy data. Furthermore, we offer a perspective on how to manage many of these potential pitfalls.

}, issn = {1477-9137}, doi = {10.1242/jcs.259656}, author = {Reiche, Michael A and Aaron, Jesse S and Boehm, Ulrike and DeSantis, Michael C and Hobson, Chad M and Khuon, Satya and Lee, Rachel M and Chew, Teng-Leong} } @article {67736, title = {Visualizing the Invisible: Advanced Optical Microscopy as a Tool to Measure Biomechanical Forces.}, journal = {Frontiers in Cell and Developmental Biology}, volume = {9}, year = {2021}, month = {2021 Sep 06}, pages = {706126}, abstract = {

The importance of mechanical force in biology is evident across diverse length scales, ranging from tissue morphogenesis during embryo development to mechanotransduction across single adhesion proteins at the cell surface. Consequently, many force measurement techniques rely on optical microscopy to measure forces being applied by cells on their environment, to visualize specimen deformations due to external forces, or even to directly apply a physical perturbation to the sample via photoablation or optogenetic tools. Recent developments in advanced microscopy offer improved approaches to enhance spatiotemporal resolution, imaging depth, and sample viability. These advances can be coupled with already existing force measurement methods to improve sensitivity, duration and speed, amongst other parameters. However, gaining access to advanced microscopy instrumentation and the expertise necessary to extract meaningful insights from these techniques is an unavoidable hurdle. In this Live Cell Imaging special issue Review, we survey common microscopy-based force measurement techniques and examine how they can be bolstered by emerging microscopy methods. We further explore challenges related to the accompanying data analysis in biomechanical studies and discuss the various resources available to tackle the global issue of technology dissemination, an important avenue for biologists to gain access to pre-commercial instruments that can be leveraged for biomechanical studies.

}, issn = {2296-634X}, doi = {10.3389/fcell.2021.706126}, author = {Hobson, Chad M and Aaron, Jesse S and Heddleston, John M and Chew, Teng-Leong} }