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Multi-Modal and Molecular Imaging of Cellular Microenvironment and Tissue Development
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Multi-Modal and Molecular Imaging of Cellular Microenvironment and Tissue Development

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Biophysics".

Deadline for manuscript submissions: closed (31 January 2021) | Viewed by 21523

Special Issue Editor

Dr. Francesco Pampaloni

E-Mail Website
Guest Editor
Buchmann Institute for Molecular Life Sciences (BMLS), Goethe-Universität Frankfurt am Main, Frankfurt am Main, Germany
Interests: light sheet microscopy; three-dimensional cell cultures; organoids for cancer research and regenerative medicine

Special Issue Information

Dear Colleagues,

Normal and pathological physiological process within the organism involve the reciprocal interaction of tissue cells with each other, with stromal cells, and with the surrounding extracellular matrix (ECM). In addition, the three-dimensional position of cells in a tissue determines the supply of oxygen, nutrients, and soluble factors, such as cytokine and hormones. Altogether, this interplay of physical and chemical signals represents the “cellular microenvironment”. The microenvironment regulates cell proliferation, motility, migration, aggregation, and differentiation. In organism development, cues from the microenvironment are essential for the differentiation of organs. Sudden or chronical alterations of the microenvironment induce a perturbation of tissue homeostasis and can lead to a pathology. For example, there is a well-proven link between an altered microenvironment, in terms of ECM composition and stromal cells population, and cancer malignancy. Visualizing cell dynamics in their three-dimensional tissue context is essential to understand the cellular and molecular processes in action. The toolbox of imaging techniques available to scientists to this aim is ample: label-free contrast, fluorescence and multi-photon microscopy, fluorescence lifetime imaging (FLIM), as well as techniques such as light sheet fluorescence microscopy and optical coherence tomography. Most of these approaches are also applicable to live imaging, which allows one to track cell motility and cell fate within tissues.

The aim of this Special Issue is presenting the most advanced developments in multi-modal microscopy and molecular imaging of the tissue microenvironment, applied to the study of the development of organisms, tissue differentiation, as well as the onset and progression of pathological processes.

Dr. Francesco Pampaloni
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • multi-modal microscopy
  • molecular imaging
  • light sheet microscopy
  • LSFM
  • tissue microenvironment
  • organ development
  • organoids

Published Papers (7 papers)

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Editorial

Jump to: Research, Review

3 pages, 180 KiB  
Editorial
Multi-Modal and Molecular Imaging of Cellular Microenvironment and Tissue Development
by Francesco Pampaloni
Int. J. Mol. Sci. 2022, 23(13), 7113; https://doi.org/10.3390/ijms23137113 - 26 Jun 2022
Cited by 1 | Viewed by 1125
Abstract
Imaging the interaction of individual cells with their surrounding tissue microenvironment is essential to advance in bioprinting, tissue engineering and cancer biology, to mention just three highly relevant fields in the life sciences [...] Full article
(This article belongs to the Special Issue Multi-Modal and Molecular Imaging of Cellular Microenvironment and Tissue Development)

Research

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13 pages, 1223 KiB  
Communication
Improvement of the Similarity Spectral Unmixing Approach for Multiplexed Two-Photon Imaging by Linear Dimension Reduction of the Mixing Matrix
by Asylkhan Rakhymzhan, Andreas Acs, Anja E. Hauser, Thomas H. Winkler and Raluca A. Niesner
Int. J. Mol. Sci. 2021, 22(11), 6046; https://doi.org/10.3390/ijms22116046 - 3 Jun 2021
Cited by 6 | Viewed by 2301
Abstract
Two-photon microscopy enables monitoring cellular dynamics and communication in complex systems, within a genuine environment, such as living tissues and, even, living organisms. Particularly, its application to understand cellular interactions in the immune system has brought unique insights into pathophysiologic processes in vivo. [...] Read more.
Two-photon microscopy enables monitoring cellular dynamics and communication in complex systems, within a genuine environment, such as living tissues and, even, living organisms. Particularly, its application to understand cellular interactions in the immune system has brought unique insights into pathophysiologic processes in vivo. Simultaneous multiplexed imaging is required to understand the dynamic orchestration of the multiple cellular and non-cellular tissue compartments defining immune responses. Here, we present an improvement of our previously developed method, which allowed us to achieve multiplexed dynamic intravital two-photon imaging, by using a synergistic strategy. This strategy combines a spectrally broad range of fluorophore emissions, a wave-mixing concept for simultaneous excitation of all targeted fluorophores, and an unmixing algorithm based on the calculation of spectral similarities with previously measured fluorophore fingerprints. The improvement of the similarity spectral unmixing algorithm here described is based on dimensionality reduction of the mixing matrix. We demonstrate its superior performance in the correct pixel-based assignment of probes to tissue compartments labeled by single fluorophores with similar spectral fingerprints, as compared to the full-dimensional similarity spectral unmixing approach. Full article
(This article belongs to the Special Issue Multi-Modal and Molecular Imaging of Cellular Microenvironment and Tissue Development)
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10 pages, 4369 KiB  
Article
In Vivo Evaluation of a Miniaturized Fluorescence Molecular Tomography (FMT) Endoscope for Breast Cancer Detection Using Targeted Nanoprobes
by Hao Yang, Weipin Qian, Lily Yang, Huikai Xie and Huabei Jiang
Int. J. Mol. Sci. 2020, 21(24), 9389; https://doi.org/10.3390/ijms21249389 - 9 Dec 2020
Cited by 6 | Viewed by 2280
Abstract
In this study, in vivo animal experiments with 12 nude mice bearing breast-cancer-patient-tissue-derived xenograft (PDX) tumors were performed aiming to verify the imaging capability of a novel miniaturized fluorescence molecular tomography (FMT) endoscope, in combination with targeted nanoparticle–near-infrared (NIR) dye conjugates. Tumor-bearing mice [...] Read more.
In this study, in vivo animal experiments with 12 nude mice bearing breast-cancer-patient-tissue-derived xenograft (PDX) tumors were performed aiming to verify the imaging capability of a novel miniaturized fluorescence molecular tomography (FMT) endoscope, in combination with targeted nanoparticle–near-infrared (NIR) dye conjugates. Tumor-bearing mice were divided into two groups by systematic injection with urokinase plasminogen activator receptor-targeted (n = 7) and nontargeted (n = 5) imaging nanoprobes as a contrast agent, respectively. Each mouse was imaged at 6, 24, and 48 h following the injection of nanoprobes using the FMT endoscope. The results show that systemic delivery of targeted nanoprobes produced a 4-fold enhancement in fluorescence signals from tumors, compared with tumors that received nontargeted nanoprobes. This study indicates that our miniaturized FMT endoscope, coupled with the targeted nanoparticle–NIR dye conjugates as a contrast agent, has high sensitivity and specificity, and thus great potential to be used for image-guided detection and removal of a primary tumor and local metastatic tumors during surgery. Full article
(This article belongs to the Special Issue Multi-Modal and Molecular Imaging of Cellular Microenvironment and Tissue Development)
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21 pages, 4803 KiB  
Article
Characterization of Mesenchymal Stem Cell Differentiation within Miniaturized 3D Scaffolds through Advanced Microscopy Techniques
by Valentina Parodi, Emanuela Jacchetti, Arianna Bresci, Benedetta Talone, Carlo M. Valensise, Roberto Osellame, Giulio Cerullo, Dario Polli and Manuela T. Raimondi
Int. J. Mol. Sci. 2020, 21(22), 8498; https://doi.org/10.3390/ijms21228498 - 11 Nov 2020
Cited by 10 | Viewed by 3497
Abstract
Three-dimensional culture systems and suitable substrates topographies demonstrated to drive stem cell fate in vitro by mechanical conditioning. For example, the Nichoid 3D scaffold remodels stem cells and shapes nuclei, thus promoting stem cell expansion and stemness maintenance. However, the mechanisms involved in [...] Read more.
Three-dimensional culture systems and suitable substrates topographies demonstrated to drive stem cell fate in vitro by mechanical conditioning. For example, the Nichoid 3D scaffold remodels stem cells and shapes nuclei, thus promoting stem cell expansion and stemness maintenance. However, the mechanisms involved in force transmission and in biochemical signaling at the basis of fate determination are not yet clear. Among the available investigation systems, confocal fluorescence microscopy using fluorescent dyes enables the observation of cell function and shape at the subcellular scale in vital and fixed conditions. Contrarily, nonlinear optical microscopy techniques, which exploit multi-photon processes, allow to study cell behavior in vital and unlabeled conditions. We apply confocal fluorescence microscopy, coherent anti-Stokes Raman scattering (CARS), and second harmonic generation (SHG) microscopy to characterize the phenotypic expression of mesenchymal stem cells (MSCs) towards adipogenic and chondrogenic differentiation inside Nichoid scaffolds, in terms of nuclear morphology and specific phenotypic products, by comparing these techniques. We demonstrate that the Nichoid maintains a rounded nuclei during expansion and differentiation, promoting MSCs adipogenic differentiation while inhibiting chondrogenesis. We show that CARS and SHG techniques are suitable for specific estimation of the lipid and collagenous content, thus overcoming the limitations of using unspecific fluorescent probes. Full article
(This article belongs to the Special Issue Multi-Modal and Molecular Imaging of Cellular Microenvironment and Tissue Development)
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25 pages, 9567 KiB  
Article
Integration of Multiple Platforms for the Analysis of Multifluorescent Marking Technology Applied to Pediatric GBM and DIPG
by Giulia Pericoli, Stefania Petrini, Ezio Giorda, Roberta Ferretti, Maria Antonietta Ajmone-Cat, Will Court, Libenzio Adrian Conti, Roberta De Simone, Paola Bencivenga, Alessia Palma, Angela Di Giannatale, Chris Jones, Andrea Carai, Angela Mastronuzzi, Emmanuel de Billy, Franco Locatelli and Maria Vinci
Int. J. Mol. Sci. 2020, 21(18), 6763; https://doi.org/10.3390/ijms21186763 - 15 Sep 2020
Cited by 11 | Viewed by 4241
Abstract
The intratumor heterogeneity represents one of the most difficult challenges for the development of effective therapies to treat pediatric glioblastoma (pGBM) and diffuse intrinsic pontine glioma (DIPG). These brain tumors are composed of heterogeneous cell subpopulations that coexist and cooperate to build a [...] Read more.
The intratumor heterogeneity represents one of the most difficult challenges for the development of effective therapies to treat pediatric glioblastoma (pGBM) and diffuse intrinsic pontine glioma (DIPG). These brain tumors are composed of heterogeneous cell subpopulations that coexist and cooperate to build a functional network responsible for their aggressive phenotype. Understanding the cellular and molecular mechanisms sustaining such network will be crucial for the identification of new therapeutic strategies. To study more in-depth these mechanisms, we sought to apply the Multifluorescent Marking Technology. We generated multifluorescent pGBM and DIPG bulk cell lines randomly expressing six different fluorescent proteins and from which we derived stable optical barcoded single cell-derived clones. In this study, we focused on the application of the Multifluorescent Marking Technology in 2D and 3D in vitro/ex vivo culture systems. We discuss how we integrated different multimodal fluorescence analysis platforms, identifying their strengths and limitations, to establish the tools that will enable further studies on the intratumor heterogeneity and interclonal interactions in pGBM and DIPG. Full article
(This article belongs to the Special Issue Multi-Modal and Molecular Imaging of Cellular Microenvironment and Tissue Development)
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14 pages, 3689 KiB  
Article
High Resolution Multimodal Photoacoustic Microscopy and Optical Coherence Tomography Visualization of Choroidal Vascular Occlusion
by Van Phuc Nguyen, Yanxiu Li, Jessica Henry, Wei Zhang, Xueding Wang and Yannis M. Paulus
Int. J. Mol. Sci. 2020, 21(18), 6508; https://doi.org/10.3390/ijms21186508 - 5 Sep 2020
Cited by 10 | Viewed by 2537
Abstract
Photoacoustic microscopy is a novel, non-ionizing, non-invasive imaging technology that evaluates tissue absorption of short-pulsed light through the sound waves emitted by the tissue and has numerous biomedical applications. In this study, a custom-built multimodal imaging system, including photoacoustic microscopy (PAM) and optical [...] Read more.
Photoacoustic microscopy is a novel, non-ionizing, non-invasive imaging technology that evaluates tissue absorption of short-pulsed light through the sound waves emitted by the tissue and has numerous biomedical applications. In this study, a custom-built multimodal imaging system, including photoacoustic microscopy (PAM) and optical coherence tomography (OCT), has been developed to evaluate choroidal vascular occlusion (CVO). CVO was performed on three living rabbits using laser photocoagulation. Longitudinal imaging of CVO was obtained using multiple imaging tools such as color fundus photography, fluorescein angiography, indocyanine green angiography (ICGA), OCT, and PAM. PAM images were acquired at different wavelengths, ranging from 532 to 700 nm. The results demonstrate that the CVO was clearly observed on PAM in both two dimensions (2D) and 3D with high resolution longitudinally over 28 days. In addition, the location and margin of the CVO were distinguished from the surrounding choroidal vasculature after the injection of ICG contrast agent. PAM imaging was achieved using a laser energy of approximately 80 nJ, which is about half of the American National Standards Institute safety limit. The proposed imaging technique may provide a potential tool for the evaluation of different chorioretinal vascular disease pathogeneses and other biological studies. Full article
(This article belongs to the Special Issue Multi-Modal and Molecular Imaging of Cellular Microenvironment and Tissue Development)
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Review

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44 pages, 1694 KiB  
Review
Molecular and Cellular Complexity of Glioma. Focus on Tumour Microenvironment and the Use of Molecular and Imaging Biomarkers to Overcome Treatment Resistance
by Silvia Valtorta, Daniela Salvatore, Paolo Rainone, Sara Belloli, Gloria Bertoli and Rosa Maria Moresco
Int. J. Mol. Sci. 2020, 21(16), 5631; https://doi.org/10.3390/ijms21165631 - 6 Aug 2020
Cited by 23 | Viewed by 4913
Abstract
This review highlights the importance and the complexity of tumour biology and microenvironment in the progression and therapy resistance of glioma. Specific gene mutations, the possible functions of several non-coding microRNAs and the intra-tumour and inter-tumour heterogeneity of cell types contribute to limit [...] Read more.
This review highlights the importance and the complexity of tumour biology and microenvironment in the progression and therapy resistance of glioma. Specific gene mutations, the possible functions of several non-coding microRNAs and the intra-tumour and inter-tumour heterogeneity of cell types contribute to limit the efficacy of the actual therapeutic options. In this scenario, identification of molecular biomarkers of response and the use of multimodal in vivo imaging and in particular the Positron Emission Tomography (PET) based molecular approach, can help identifying glioma features and the modifications occurring during therapy at a regional level. Indeed, a better understanding of tumor heterogeneity and the development of diagnostic procedures can favor the identification of a cluster of patients for personalized medicine in order to improve the survival and their quality of life. Full article
(This article belongs to the Special Issue Multi-Modal and Molecular Imaging of Cellular Microenvironment and Tissue Development)
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