- Sony Biotechnology
- Sony Biotechnology
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.
Listen to Dr. Feng Zhang who was first to adapt CRISPR-Cas9 for genome editing on eukaryotic cells explain CRISPR:
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.
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.
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.
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...
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.
T-cells are part of the adaptive immune system with several subtypes including CD8+ effector T-cells and CD4+ helper T-cells. Main categories can be divided into several sub-categories. What are the critical markers to study T-cells? From the key subsets how can other subsets be defined?
Thank your super hero mom this Mother’s Day by giving her some quality time and something to remember—the gift of knowing about the work you do that makes the world a better place. To help you with what could be the best little talk ever here’s a 5-minute animation from the Royal Society that explains gene editing and the importance CRISPR-Cas-9 we hope you will find useful. Happy Mother’s Day to all, thank you for what you do. Video: What is gene editing and how does it work?
Tips for choosing the best clone for flow cytometry experiments to improve research quality and save time and money.