captcha-code refresh-capcha

    Forgot Password

    About Sony Biotechnology

    • Enabling flow cytometry analysis of tissues with spectral flow cytometry

      23 Jun 2017

      Flow cytometry has long been considered a tool of hematologists and immunologists who primarily work with the hematopoetic system.  Flow cytometry is capable of performing the simultaneous detection of more than 20 parameters; however it requires the sample to be prepared into a single cell suspension. The need for a single cell suspension makes the sample preparation more intensive for cells derived from tissues.  Many tissues are also highly autofluorescent, which results in background noise that limits detection of low levels of expression on conventional flow cytometers.

      Researchers from the Institut Pasteur, Immunology Department in Paris France have published a study in PLoS One1 which simultaneously measured 21 parameters (19 fluorochromes) on cells derived from mouse embryonic heart and intestine. Both tissue types contain autofluorescent cells. With conventional cytometers, specific band pass filters must be carefully selected to enable compensation for spectral overlap.  The more overlap between fluorescent dyes, the more difficult to perform conventional compensation. Furthermore, cells such as those described in the article have larger amounts of inherent autofluorescence complicating the data analysis. With the SP6800 a spectral unmixing algorithm is used to determine the abundance of signal from each fluorochrome. This allows for the separation of highly overlapping signals including GFP and FITC as well as effective management of autofluorescence. Directly comparing results on two conventional flow cytometers and the SP6800, only the SP6800 was capable of effective phenotypic resolution of the target populations.

      REFERENCES

      1 Schmutz S, Valente M, Cumano A, Novault S (2016) Spectral Cytometry Has Unique Properties Allowing Multicolor Analysis of Cell Suspensions Isolated from Solid Tissues. PLoS ONE 11(8): e0159961. journals.plos.org

      More Less

    • Learning more about CRISPR from experts

      23 Jun 2017

      Listen to Dr. Feng Zhang who was first to adapt CRISPR-Cas9 for genome editing on eukaryotic cells explain CRISPR in this 2 minute video: Questions And Answers About CRISPR

      Today Dr. Zhang leverages CRISPR and other methodologies to study the role of genetic and epigenetic mechanisms underlying diseases, specifically focusing on disorders of the nervous system. He is especially interested in complex disorders, such as psychiatric and neurological diseases, that are caused by multiple genetic and environmental risk factors and which are difficult to model using conventional methods. His lab is focused on developing novel tools to better understand and treat neuropsychiatric diseases and applying these novel tools to interrogate gene function in animal and stem cell models.  Zhang’s methods are also being used in the fields of immunology, clinical medicine, cancer biology, and other areas of research. Zhang’s long-term goal is to develop novel therapeutic strategies for disease treatment.

      Dr. Zhang is the James and Patricia Poitras Professor in Neuroscience at the McGovern Institute for Brain Research and in the departments of Brain and Cognitive Sciences and Biological Engineering at the Massachusetts Institute of Technology (MIT). He also has appointment with the Broad Institute of MIT and Harvard.

      More Less

    • Sorting iPSC derived Cardiomyocytes using the SH800 Cell Sorter

      15 Jun 2017

      Pluripotent stem cells (PSCs) offer an unprecedented opportunity for both disease modeling and personalized medicine. In particular, PSC derived cardiomyocytes (CM) mature into adult cardiomyocytes when transplanted into neonatal rat hearts allowing iPSC modeling of cardiomyopathy. A recent published study by Kwong et al 1 shows the successful isolation of cardiac progenitor cells (CPCs) from mouse embryonic stem cells. To achieve this, a mESC line expressing Isl1-Cre; Rosa-RFP; aMHC-GFP was generated. The expression of Rosa-RFP allowed tracking of CPCs destined to mature into CM. RFP+ CPCs were sorted using the Sony SH800 cell sorter using the 130um microfluidics sorting chip.

      The growth and cell health of these sorted cells was monitored in-vitro by microscopy prior to injecting them in nude mice. All sorted cells were viable and differentiated into adult CMs in the model animal.

      Such an approach may be extended for generating other types of adult cells prone to disease, such as skeletal muscle cells, pancreatic cells, and renal cells, from hiPSCs to study and model adult-onset human diseases.

       

      Reference

      1. Neonatal Transplantation Confers Maturation of PSC-Derived Cardiomyocytes Conducive to Modeling Cardiomyopathy. Cell Reports 2017, 18: 571–582. Cho GS, Lee DI, Tampakakis E, Murphy S, Andersen P, Uosaki H, Chelko S, Chakir K, Hong I, Seo K, Chen HV, Chen X, Basso C, Houser SR, Tomaselli GF, O’Rourke B, Judge DP, Kass DA, and Kwon C
      More Less

    • ISSCR June 14-17: See us at booth #535

      15 Jun 2017

      We’re still in Boston and you’ll find us in booth #535, at ISSCR where researchers from around the world will discuss advanced stem cell science and regenerative medicine.

      At our booth you can get some 1:1 time with the FX500 Replaceable Fluidics Cell Sorter. Seeing the system up close will give you a good feel for how easy it is to learn and use. The system features automation that handles many common tasks from start up through sort monitoring and shut down. Software also guides users through the replacement of single use consumables including sorting chip, sheath line and sample line to support your aseptic workflow.

      ISSCR has always been a conference for visionaries. This year opening the Presidential Symposium, A Decade of Human iPSCs from Discovery to Clinic. world renowned scientists Sally Temple (Neural Stem Cell institute) Magdalena Zernicka-Goetz (University of Cambridge), Rudolf Jaenisch, (Whitehead Biomedical Research Institute), Shinya Yamanaka (Center for IPS Cell Research & Application) and Joanna Wysocka (Stanford) take a look at how far we have come and the path forward.

      More Less

    • Using flow cytometry to evaluate compatibility of tyrosine kinase inhibitors with immunotherapy

      05 Jun 2017

      Immunotherapy has shown great promise as a cancer treatment resulting in several FDA approved drugs and clinical trials

      For example, a promising therapy in several tumors is the blocking of the PD-1/PD-L1 interaction that is over expressed on many types of tumor cells.  However, success depends on the patient’s immune system to respond to the tumor.

      Immunotherapy is typically used in combination with other agents. The most common combination are chemotherapeutic agents, which are known for suppressing the immune system. These agents include tyrosine kinase inhibitors (TKIs). TKIs are used to treat several neoplastic diseases, most notably chronic myelogenous leukemia (CML).  To further examine the effect of TKIs on the immune system, Heine1 et al., studied the effect of several TKIs on the induction of myeloid derived suppressor cells (MDSCs). MDSCs are negative regulators of the immune system that can accumulate in cancer patients.

      In this study, researchers used a Sony SH800 cell sorter to isolate CD14+ cells (monocytes) and CD8+ T cells from healthy volunteers and patient samples. The monocytes were co-cultured with hepatic stellate cells (HSCs) to induce MDSCs. The co-cultures were exposed to several TKIs including: nilotinib, dasatinib, sorafenib, and sunitinib at different times points. Cells were characterized in several assays including apoptosis and immunophenotyping on the Sony SP6800 Spectral Analyzer. When the monocytes were treated with three out of the four agents, there was decrease the differentiation of monocytes into MDSCs.  T-cell suppression was also reduced.

      These results suggest that the timing and choice of TKI can modulate the success of cancer immunotherapeutics. This study may be a useful model to examine the impact of other therapeutics on the immune system and the compatibility of those therapeutics with immune checkpoint inhibitors.

      1Heine, Annkristin, et al. "The induction of human myeloid derived suppressor cells through hepatic stellate cells is dose-dependently inhibited by the tyrosine kinase inhibitors nilotinib, dasatinib and sorafenib, but not sunitinib." Cancer Immunology, Immunotherapy 65.3 (2016): 273-282.

      More Less

    • Join us at booth 502 for CYTO 2017

      02 Jun 2017

      Please join us at CYTO 2017 booth #502 June 10-14 for a walk through on how spectral technology provides better data for research involving heterogeneous samples. You’ll also be able to get hands-on experience with new DeNovo 6.0 software that now natively supports spectral analysis. We’ll also be showing our best in class SH800 cell sorter and our FX500 replaceable fluidics cell sorter. The FX500 uses single use consumables to support an aseptic workflow with automation that simplifies operation and enables walk-away sorting.

      Our commercial tutorial will be presented in room 309 on Monday June 12 from 12:30 to 13:30. This year’s presentation will focus on how raw spectral data provides a more accurate and unbiased data for clustering transformation and analysis. We will show you data that demonstrates how these transformations make it easier to see populations in heterogeneous samples using spectral flow cytometry.

      As you probably know, this year’s CYTO scientific program promises to be one of the most interesting in memory focusing new approaches for deeper understanding of biological systems. In her opening lecture, Illuminating Biology at the Nanoscale with Single-Molecule and Super-Resolution Imaging, Dr. Xiaowei Zhuang, from the David B. Arnold Professor of Science at Harvard University and an investigator of Howard Hughes Medical Institute, will discuss single-molecule and super-resolution imaging methods developed by her lab apply to biological studies.

      This deep look at optical tools continues with the lecture from Ed Boyden, professor of Biological Engineering and Brain and Cognitive Sciences at the MIT Media Lab and the MIT McGovern Institute. His lecture, Optical Tools for Understanding Biological Systems focuses on the work performed at the Synthetic Neurobiology Group, he leads.  Tools developed by the group are designed to analyze and repair complex biological systems, such as the brain, to reveal ground truth principles of biological function of these systems and to repair them.

      Whether you are attending CYTO2017 to network or examine new approaches to understanding disease and improving clinical care, CYTO is the must attend event for idea sharing and innovation. We look forward to seeing you there.

      More Less

    • Standardization In - Variability Out – Reliable Data from your Cell Analyzer

      25 May 2017

      As most researchers know, all instruments have some variability and over time settings continue to drift. Obviously, instrument variability is “bad” because it can unwittingly impact your data – forcing you to re-run experiments to determine what caused data anomalies.

      To eliminate instrument variability, Sony cell analyzers include a new Standardization function that sets Sony systems to a global specification with one simple click.  Behind the scenes, Sony Standardization Mode automatically sets universal parameters for laser alignment and calibration; detector correction and dark noise elimination.  This ensures high reliability.  When working with a single instrument, standardization ensures you’ll get the same specification from day to day. When working across multiple instruments, standardization ensures all instruments are set to the same specification.  Simple as that.

      In conventional flow, instrument QC is performed to verify instrument performance.   Beads determine bandpass filter settings and laser alignment and laser calibration are performed manually.  Although a comprehensive validation is performed, there is no way to verify that settings from one instrument match another and no guarantee that the operator has setup the instrument correctly.

      To demonstrate the capabilities of the standardization function, we gathered data from six SA3800 spectral analyzers located in the United States, Europe and Japan.  To see how standardization produced reproducible results, download the technical note, Standardization capability on a Sony spectral flow cytometer.

      More Less

    • Using cell sorting to isolate cells with genes edited by CRISPR-Cas9

      19 May 2017

      With the ability to easily identify and isolate single cells from a heterogeneous phenotypic population, a cell sorter is a useful tool for laboratories studying and applying the CRISPR technology as a tool for editing genes and creating single cell libraries.

      CRISPR genome editing tools can be used to remove, change or insert DNA sequences into genes. The resulting mutations are maintained in the genome as cells proliferate –in contrast to transient transfection methods.

      Edits to genes are accomplished by directing the Cas9 nuclease to cleave DNA at a specific sequence and a guide RNA (gRNA) into the cell through transfection (or electroporation). For the edit to be completed the cell must be successfully transfected and the DNA cleaved by the Cas9 (referred to as cleavage efficiency).

      Many factors can impact cleavage efficiency including the efficiency of the transfection. A commonly used method to identify cells that have been successfully transfected is to include a plasmid encoding for a fluorescent protein such as eGFP in the transfection along with Cas9 and gRNA.

      Cells can then be sorted based on the expression of the fluorescent protein. The SH800 sorter, lets researchers deposit target cells into 96- or 384 multi-well devices and indexing software records the X and Y well position of each cell. The software can also correlate the fluorescence and scatter phenotype of sorted cells for meta-data analysis.

      The Sony SH800 has been used by several prominent research laboratories to optimize CRISPR expressing cells and to create CRISPR variants with improved function. For more information download this Application Data that demonstrates how the Sony SH800S can be used for single cell sorting of CRISPR/CAS 9 expressing cells.

      Other laboratories have used the SH800 for routine sorting of CRISPR modified cells. Learn more about sorting CRISPR modified cells in the publications below:

      Oakes, Benjamin L., et al. "Profiling of engineering hotspots identifies an allosteric CRISPR-Cas9 switch." Nature biotechnology (2016).

      Sluch, Valentin M., et al. "Differentiation of human ESCs to retinal ganglion cells using a CRISPR engineered reporter cell line." Scientific reports 5 (2015).

      Goto, Teppei, et al. "Hypomorphic phenotype of Foxn1 gene-modified rats by CRISPR/Cas9 system." Transgenic research (2016): 1-12.

      Oakes, Benjamin L., Dana C. Nadler, and David F. Savage. "Protein engineering of Cas9 for enhanced function." Methods in enzymology 546 (2014): 491.

      Schrage, Ramona, et al. "The experimental power of FR900359 to study Gq-regulated biological processes." Nature communications 6 (2015).

      Hayashi, Masayasu, et al. "Chd5 Regulates MuERV‐L/MERVL Expression in Mouse Embryonic Stem Cells Via H3K27me3 Modification and Histone H3. 1/H3. 2." Journal of Cellular Biochemistry (2015).

      More Less

    • How the immune system impacts liver regeneration

      17 May 2017

      Unlike most organs in the human body the liver is able to regenerate allowing donors and patients to re-grow resected tissue. The success and rate of regeneration can be negatively impacted by several factors including sub-septic conditions. One factor is LFA-1, an integrin lymphocyte function-associated antigen-1.

      To better understand how LFA-1 affect liver regeneration, Jörger, et al.1 used a mouse model to determine the functional role of immune cells, especially NKT cells, in response to partial hepatectomy. In this study lymphocytes were characterized on the Sony SP6800 spectral analyzer to determine innate and adaptive immune cells in liver regeneration.  In addition, the study determined that NKT lymphocytes were examined to determine their ability to play a protective role in liver regeneration.

      References

      1. Jörger A-K, Liu L, Fehlner K, Weisser T, Cheng Z, Lu M, et al. (2016) Impact of NKT Cells and LFA-1 on Liver Regeneration under Subseptic Conditions. PLoS ONE 11(12): e0168001. doi.org
      More Less

    • Immunology 2017 Washington, D.C. May 13-17

      09 May 2017

      Stop by booth #319 at Immunology 2017 for a walk through on how spectral technology, the base of our cell Analyzers, provides better data for research involving heterogeneous samples. You can also get some “face-to-face” time with our newest cell sorter, the FX500 Replaceable Fluidics Cell Sorter. See how the system’s single use consumables support an aseptic workflow and how automation enables walk away operation.

      From May 13 – 17, Immunology 2017 in Washington D.C., is expected to be the largest gathering of immunologists worldwide. The scientific program is packed with presentations from leading immunologists on a full range of topics to stimulate discussion and offer new tools for your research.

      For more information visit Immunology 2017

      More Less