From 3D organs to single-celled - Perkin Elmer invites you to attend the 2020 Annual Meeting of the China Cell Biology Society.

Cell 3D model culture is better able to simulate microenvironments, intercellular interactions, and invivia biological processes.
provides more physiologically relevant conditions than biochemical detection and 2D models.
, they are more morphologically and functionally divided, which also gives them a closer look at the characteristics of cells in the body.
more and more researchers are now applying 3D cell culture, microtissue and organ-like techniques to bridge the gap between 2D cell culture and animal models in the body.
especially the research and use of organ-like organs (Organoid) is an in vitro derived 3D cell aggregate derived from stem cells, with similar organ structure and function.
in recent years, 3D organ culture technology has matured and is becoming an important model for drug screening, personalized treatment and development al-development research.
, however, the 3D culture technology of cells faces many challenges: first, it is difficult to cultivate consistent, reproducible 3D microtissues, especially organ-like cultures;
for 3D microtissue samples, imaging with traditional frozen slicing staining or direct lying with confocal microscopes has many challenges: frozen slicing imaging does not provide all the information about stereoscopic samples, especially cell location information on the Z axis; In addition, the large number of images obtained by these two methods of photography requires the analysis and statistics of their data with the help of other analytical software, the analysis flux is very low, and importantly, both methods are slow, low in flux, and the scanning time of a 3D microorganized scan for several hours, greatly limiting the development of 3D microtissue research.
high-content cell imaging can, on the premise of maintaining cell structure and functional integrity, carry out a comprehensive scan of fluorescence of cells and subcellular levels, detect cell morphology, growth, differentiation, migration, apoptosis, metabolic pathways and signal transduction, and obtain a large amount of relevant information in a single experiment.
have been used well in cell apoptosis, cell cycle, cytotoxicity, receptor protein translocation, protein interaction, etc., and have been proved to be an important research tool in cell biology, cancer research, pathogen biology, drug research, systems biology, cardiovascular disease research, stem cell research, nerve cell research and other fields.
high-content cell imaging analysis system provided by Perkin Elmer, which uses Nipkow turntable scanning technology with high sensitivity sCMOS detectors, can quickly capture biological processes occurring in cells, but also because of its reduced photobleaching and phototoxicity characteristics, combined with flood-soaked high-value aperture objectives, can achieve high-volume confocal high-resolution imaging of living cells, small-scale organisms and 3D microtissue samples.
combined with powerful Harmony analytics software, it is possible to conduct group statistical analysis of complex types at the cell and subcellular levels.
the system has a wide range of applications in the field of cell biology.
Perkin Elmer's 3D solutions are not limited to 3D micro-tissues, including model organisms, cell pseudo-foot and other stereostructures can be completed through high-content systems to complete comprehensive detection and analysis: Perkin Elmer's single-cell ICP-MS technology, based on the industry's faster cells Pulse signal reading speed (up to 100,000 points per second) can quantify the metal and nanoparticle content of a single cell as low as Ak-level, and determine the distribution of metal mass and the number of metal-containing cells in the cell group to assess and quantify the heterogeneity of the cell group.
is applicable to the in-depth study of human, animal, plant and other tissue and organ cells.
for example, metal-containing drugs and nanoparticles are increasingly widely used in cancer treatment and detection, single-cell ICP-MS can be finely tracked to understand the absorption and metabolism of the disease at the cellular level of the diseased tissue, help to understand the mechanism of cancer and improve treatment levels.
the copper content in the cisplatin intake organisms with varying cisplatin intake over time is strictly regulated by a very effective and complex steady-state mechanism that controls the absorption, distribution and discharge of elements by two ovarian cancer cell lines, A2780 (cisplatin sensitive) and A2780/CP70 (cisplatin drug-resistant) over time.
the current data obtained by the cell copper steady state model is only a "skeleton", using SC-ICP-MS to measure the copper (Cu) content in peripheral blood mononuclear cells (PBMC), to understand the disorder or imbalance of the steady state mechanism may lead to biological dysfunction, and may provide an effective means of further research with certain diseases (e.g. inflammation, asthma, aging process, cancer, etc.). Examples of copper content applications in peripheral blood mononucleocells (PBMC) in
: The current applicationof of 3D microtissue organs is mainly concentrated in tumor research (drug sieve models, drug sieves, tumor immunity, personalized medicine), stem cell and developmental biology, in vitro model research (infection models, toxicity evaluation), materials and feeding Drug Research and others: Oncology Research On June 17, 2019, the journal Cell and Disease published online the results of the study of the Qian's military research group, modified CAR T cells targeting the proximal epitope of mesothelin enhances the anti-body function ss big solid tumor.
work on optimizing tumor CAR T immunotherapy.
MSLN (Mesothelin), a seductive antigen for the treatment of chimeric antigen receptor (CAR) T, the selection of epitope in MSLN is critical.
in the study, the authors constructed two types of CAR systems for MSLN's I-zone (meso1 CAR, also known as the far end region of the membrane) or the MESo3 CAR (meso3 CAR, also known as the membrane near end region) using modified piggyBac transposons.
, meso3 CAR T cells express higher levels of CD107 alpha after activation and produce higher levels of interleukin 2, TNF-alpha, and IFN-in vitro for cancer cells that express multiple MSLNs. After
, the authors constructed a 3D tumor cell model for gastric cancer and ovarian cancer, and used the model to test the two CAR T systems, and completed the imaging and analysis of the 3D tumor CART killing system through perkin Elmer Opera Phenix high-content system, which proved that at 3D cell level, meso3 CAR T cells had a higher lethal effect than meso1 CAR T cells.
follow-up study, the authors further validated the mouse model of gastric cancer NSG with the help of perkin Elmer Xenogen IVIS imaging system, and also demonstrated that meso3 CAR T cell-mediated antitumor response was stronger than that of meso1 CAR T cells.
we further determined that meso3 CAR T cells can effectively inhibit the growth of large ovarian tumors in the body.
in general, this study proves that meso3 CAR T-cell therapy has better immunotherapy than meso1 CAR T cell therapy in the treatment of MSLN-positive solid tumors, and provides new and effective CAR T therapy for the immunotherapy of solid tumors.
Stem Cell and Developmental Biology In November 2018, the University of Glasgow's Cancer Science Research Agency, Nature Communications, published an article entitled "The Phospholipid PI (3,4)P 2 Is 2 Is an Apical Identity Identity" In which the polarization mechanism of epithelial cells was studied, using MDCK cysts as a model, and eventually finding that PI (3,4) P2 phosphase is an important molecule that determines the polarization of upper cell sder.
in this article, the authors first found that the distribution of phosphate modifier PI (3,4)P2 is critical in the process of polarization of epithelial cells, and then, they use the distribution of PI (3,4) P2 as phenotypes to screen which proteins are knocked out to affect their distribution.
the process is achieved through Perkin Elmer's Opera Phenix high-content system, the author first through the high-content system of pre-scan imaging function of the microsphere intelligent layer-cutting scanning, select the cross-section of the large layer, and then the cell region, sub-cell nuclei, cytoplasm, inner, outer and cell connections, and so on, and then calculate the fluorescence intensity of each region.
the authors used this method to analyze the distribution of phosphatase in some mutated microspheres, and found significant positioning changes when important upstream proteins, such as PIP proteins, were knocked off. In addition to
, the author also used the high-content system to analyze the cavity phenotype of microspheres, MDCK cystcontainas containing how many cavities directly reflect whether its function is normal, only the normal polarization of cysts can have a normal cavity.
the same, the authors used high-content pre-scan imaging to scan all balls in layers, select the cavities, press them together, and then algorithmly select the cavities to analyze their number.
the authors also used this method to screen a series of genes, screening several genes that significantly affect cavity formation, and subsequently clarified their regulatory mechanisms.
In vitro Model Research - Liver Injury Model 2018, Wang's team published a cover article in the new journal Advanced Biosystems, which demonstrates a new drug-based liver injury research model, the LBS Microliver Model (Liver Biomatrice Scaffolds, LBSs).
the model introduces a natural decellular liver stent based on HepaRG cells, which can be used for long-term 3D culture of liver cells.
the new model has higher physiological correlation and toxicological prediction sensitivity in the specific environment of liver tissue provided by LBS.
the authors used perkin Elmer Operetta CLS high-content screening system to further assess the hepatotoxicity of eight antidepressants.
combined with specific dye combinations, the effects of drug treatment on microliver models were detected in terms of cell vitality, apoptosis, bile accumulation, fat change, oxidative stress and mitochondrial toxicity. Many of these parameters
use complex high-content analysis methods.
results show that the LBS microhepatosphere model can be highly specific to predict drug hepatotoxicity and assist in further toxicology mechanism research.
study also used Perkin Elmer's Engisht multi-functional imaging enzyme marker to track changes in cell activity under different cultures using The Alamar Blue method. the molecular and cellular level detection program provided by
Perkin Elmer runs through the course of drug hepatotoxicity research in this paper.
from cell proliferation and enzyme activity analysis of microliver models to functional verification and toxicology multi-indicator analysis of 3D models, Perkin Elmer can provide targeted applications.
Materials and Administration Research June 2019, the School of Biology and Environmental Sciences at University College Dublin, Ireland, and the Conway Institute published an article in Small magazine entitled "A High-Sov Automatscopy Platform for Quantitative Phey ping of Nano Uptake and Transport in Spoids".
the study used Perkin Elmer's high-content Opera Phenix system to build a complete model for the study of nanocarrier ingestion and transportation at the 3D micro-tissue level.
the author first carried out the optimization of 3D microtissue culture and high-content photography, mainly studied the influence of culture conditions and fixed methods on different concentrations of matrix glue, and determined the culture method, fixed method and matrix glue concentration and dosage according to the results of the experiment.
, the authors also examined the high-content shooting quality by dyeing the distribution of trans to trans Gorky markers (GM130, GalT and TGN46), proving that perkin Elmer's high-content system does have a very high resolution and that it is sufficient to study the intake of nanoparticles.
next, the author reconstructs the layer-cut scanning pictures through Harmony software, obtains a large brightness projection and a 3D reconstruction view, and quantitatively measures NP absorption and penetration in the spherical sphere. After
, the authors selected proteins that function in nanoparticle cytoswallowation and used RNAi silence for potential gene screening to determine that the model could be used to evaluate the ingestion and transport of 3D microsphere NP.
more details, you are welcome to attend the luncheon held on August 4th at the 2020 National Conference of the Chinese Society of Cell Biology, dry goods reports, lunch gifts, surprise gifts and more.
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