Flow and image cytometry side by side for the new frontiers in quantitative single‐cell analysis

Tárnok, Attila

doi:10.1002/cyto.a.20709

Cited by 5 publications

(5 citation statements)

References 26 publications

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“…The identification of new structures involved in cell-to-cell communication has led to the development of new analytical and imaging tools, which have allowed us to enhance our understanding of the ubiquitous phenomenon of cell-to-cell communication. Such tools include quantitative cell microscopy, now mainly represented by two distinct techniques: flow cytometry (FCM) and image cytometry (IC) (5). FCM allows analysis of cell suspensions, cell-by-cell quantification of optical signals, rapid analysis (several hundred cells per second), and cell sorting, whereas IC allows precise localization and quantification of optical signals emitted by each point of the observed field and image analysis (morphology and morphometry).…”

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confidence: 99%

Flow cytometric analyses disclose intercellular communications in FasL‐stimulated T cells: Results and trouble shooting

et al. 2011

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LIFE depends on the ability of cells to communicate with one another. Much of this crosstalk occurs at cell-cell contacts and is regulated by complex structural interface: neurological and immunological synapses that transmit cell-cell signals through the extracellular space, relying on mechanisms of ligand-receptor signalling across tight cell-cell junctions. In addition to these well-known examples, other cellular structures involved in cell-cell communication have been identified (1). A frequent result of cell-cell communication is collective population behavior that can potentially lead to complex phenotypes not observed in separate individual cells, for example, in development or tumor formation (2,3). Immune responses against pathogens or any foreign antigens require fine immune regulation, where cellular communications are mediated by either soluble or cell surface molecules. Generally, absorption, exosome production and uptake, internalization, and membrane nanotube formation are the probable mechanisms through which the membrane fragments and cytoplasmic content are transferred from one cell to another (4). The identification of new structures involved in cell-to-cell communication has led to the development of new analytical and imaging tools, which have allowed us to enhance our understanding of the ubiquitous phenomenon of cell-to-cell communication. Such tools include quantitative cell microscopy, now mainly represented by two distinct techniques: flow cytometry (FCM) and image cytometry (IC) (5). FCM allows analysis of cell suspensions, cell-by-cell quantification of optical signals, rapid analysis (several hundred cells per second), and cell sorting, whereas IC allows precise localization and quantification of optical signals emitted by each point of the observed field and image analysis (morphology and morphometry). FASL STIMULATION OF T CELLSThe Fas/CD95 surface receptor mediates rapid death of various cell types, including autoreactive T cells, which can trigger autoimmunity. In this study we investigate novel aspects of Fas signaling that define a ''social'' dimension to receptor-induced apoptosis. At least in some cell types, FasL can be stored in granules and then rapidly released at the cell surface in response to external stimuli. Alternatively, FasL can be cleaved from the membrane by matrix metalloproteases and act as a soluble cytokine (6). In its membrane-bound form, FasL acts as a ligand for Fas and can trigger Fas-induced death.In our experiments, CD41 T cells were purified from PBMC by negative selection using the MACS system. CD41 T cells and Jurkat cells were both treated with FasL at a final concentration of 0.5 lg/ml for a maximum of 2 h. Our previous findings (7) showed that Fas stimulation led to an intriguing (and previously unsuspected) asymmetry of death propagation according to cell conjugation and rapidly induced extensive membrane nanotube formation between neighboring T cells. The experiments were performed using flow cytometric (FC) and morphologic (fluorescence and ti...

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confidence: 99%

Flow cytometric analyses disclose intercellular communications in FasL‐stimulated T cells: Results and trouble shooting

et al. 2011

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“…Cell membranes extracted from red blood cells (RBCs), white blood cells, platelets, bacteria, cancer cells and stem cells have been used in the previous studies 12–14 . In particular, RBCs have a lifespan of ~120 days before their immune-suppression by the body 15,16 . Phagocytosis of NPs uniformly coated by RBC membrane (Rm) is reduced by the expressed cluster of differentiation 47 (CD47) on the membrane 17–21 .…”

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confidence: 99%

Red Blood Cell Membrane Bioengineered Zr-89 Labelled Hollow Mesoporous Silica Nanosphere for Overcoming Phagocytosis

Lee

Vyas

Kim

et al. 2019

Sci Rep

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Biomimetic nanoparticles (NPs) have been actively studied for their biological compatibility due to its distinguished abilities viz. long-term circulation, low toxicity, ease for surface modification, and its ability to avoid phagocytosis of NPs by macrophages. Coating the NPs with a variety of cell membranes bearing the immune control proteins increases drug efficacy while complementing the intrinsic advantages of the NPs. In this study, efforts were made to introduce oxophilic radiometal 89 Zr with hollow mesoporous silica nanospheres (HMSNs) having abundant silanol groups and were bioengineered with red blood cell membrane (Rm) having cluster of differentiation 47 (CD47) protein to evaluate its long-term in vivo behavior. We were successful in demonstrating the increased in vivo stability of synthesized Rm-camouflaged, 89 Zr-labelled HMSNs with the markedly reduced 89 Zr release. Rm camouflaged 89 Zr-HMSNs effectively accumulated in the tumor by avoiding phagocytosis of macrophages. In addition, re-injecting the Rm isolated using the blood of the same animal helped to overcome the immune barrier. This novel strategy can be applied extensively to identify the long-term in vivo behavior of nano-drugs while enhancing their biocompatibility.

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“…In summary, this issue emphasizes that, as noted over the last few years (1, 2), imaging and particularly image analysis combined with classification algorithms have become an integral part of the wealth of science and knowledge within the cytometry community. Cytometry Part A is increasingly recognized as a home for image‐based cutting edge research and development, as witnessed by the proportion of these publications in this focus issue.…”

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confidence: 98%

“…Furthermore, it is a great challenge to quantify cell morphology and intracellular structures either in three or even four dimensions given the great heterogeneity between cells and the heterogeneity in quality between images and samples. These important aspects of a quantitative and unbiased cell analysis for cytomics or cell systems analysis have been frequent issues in the recent past (1, 2).…”

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Tárnok

2010

Cytometry Pt A

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Flow and image cytometry side by side for the new frontiers in quantitative single‐cell analysis

Cited by 5 publications

References 26 publications

Flow cytometric analyses disclose intercellular communications in FasL‐stimulated T cells: Results and trouble shooting

Flow cytometric analyses disclose intercellular communications in FasL‐stimulated T cells: Results and trouble shooting

Red Blood Cell Membrane Bioengineered Zr-89 Labelled Hollow Mesoporous Silica Nanosphere for Overcoming Phagocytosis

Telling one from another

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