Optical imaging is a valuable tool to visualise biological processes in the context of the tissue. Each imaging modality provides the biologist with different types of information - cell dynamics and migration over time can be tracked with time-lapse imaging (e.g. intra-vital imaging); an overview of whole tissues can be acquired using optical clearing in conjunction with light sheet microscopy; finer details such as cellular morphology and fine nerve tortuosity can be imaged at higher resolution using the confocal microscope. Multi-modal imaging combined with image cytometry - a form of quantitative analysis of image datasets - provides an objective basis for comparing between sample groups. Here, we provide an overview of technical aspects to look out for in an image cytometry workflow, and discuss issues related to sample preparation, image post-processing and analysis for intra-vital and whole organ imaging.
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http://dx.doi.org/10.1016/j.cellimm.2020.104086 | DOI Listing |
J Biomed Opt
January 2025
Tel Aviv University, Department of Biomedical Engineering, Faculty of Engineering, Tel Aviv, Israel.
Significance: Imaging flow cytometry allows highly informative multi-point cell analysis for biological assays and medical diagnosis. Rapid processing of the imaged cells during flow allows real-time classification and sorting of the cells. Off-axis holography enables imaging flow cytometry without chemical cell staining but requires digital processing to the optical path delay profile for each frame before the cells can be classified, which slows down the overall processing throughput.
View Article and Find Full Text PDFEur J Pharmacol
January 2025
State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, Jiangsu 211198, PR China. Electronic address:
Fc receptor γ subunit (FcRγ) activation plays a crucial role in cancer carcinogenesis. Here, we aimed to uncover the impact of FcRγ on circulating tumor cells (CTC) colonization and the underlying mechanism. FcRγ deficient (FcRγ) mice were used to investigate the functional effects of FcRγ in cancer metastasis, and the results demonstrated that FcRγ deficiency significantly promotes metastasis.
View Article and Find Full Text PDFMethods Cell Biol
January 2025
Pathology, Leiden University Medical Center, Leiden, The Netherlands. Electronic address:
In recent years, significant advancements have been achieved in the development of multiplex imaging methodologies for immunophenotyping, enabling a comprehensive characterization of the complexity of tumor microenvironments. Imaging mass cytometry combines the detection of over 40 cellular targets with spatial information, enabling the identification of not only which cells are present in a tissue but also their localization relative to each other. Here, we present an easy-to-implement imaging mass cytometry workflow that ranges from antibody selection and testing to running a full panel.
View Article and Find Full Text PDFBiomater Adv
December 2024
AO Research Institute Davos, Clavadelerstrasse 8, Davos 7270, Switzerland.
The immunomodulatory properties of hyaluronan and its derivatives are key to their use in medicine and tissue engineering. In this work we evaluated the capability of soluble tyramine-modified hyaluronan (THA) synthesized from hyaluronan of two molecular weights (low M = 280 kDa and high M = 1640 kDa) for polarization of THP-1 and peripheral blood mononuclear cells (PBMCs)-derived macrophages (MΦs). We demonstrate the polarization effects of the supplemented THA by flow cytometry and bead-based multiplex immunoassay for the THP-1 derived MΦs and by semi-automated image analysis from confocal microscopy, immunofluorescent staining utilizing CD68 and CD206 surface markers, RT-qPCR gene expression analysis, as well as using the enzyme-linked immunosorbent assay (ELISA) for PBMCs-derived MΦs.
View Article and Find Full Text PDFCell Biochem Biophys
January 2025
Department of Pharmacy, The Fourth Affiliated Hospital of Soochow University, Jiangsu, Suzhou, 215000, China.
Total glucosides of paeony (TGP) have been investigated for their effects on cardiomyocyte hypertrophy induced by angiotensin II (Ang II). In this study, rat cardiomyocyte H9c2 cells were treated with various doses of TGP (0, 12.5, 25, 50, 100, 200, and 400 μmol/L), and cell viability was assessed using the MTT method to determine an optimal dose.
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