BD ACCURI NEWS
The BD FACSMelody™ cell sorter in biopharmaceutical R&D
Mark Gilbert manages the core flow cytometry facility at NGM Bio, a research-driven biopharmaceutical company headquartered in South San Francisco, CA. Last fall, NGM Bio’s cell line development group installed a new BD FACSMelody™ cell sorter. Dr. Gilbert spoke to us about why the group needed its own sorter and why the BD FACSMelody is boring—in a good way!
Read the interview »
Automating dose-response cell cycle analysis
Flow cytometry is especially well-suited for dose-response studies because it can rapidly and accurately assess multiple markers simultaneously. In a single assay, you can assess a compound’s effects on cell cycle, cellular viability, apoptosis, mitochondrial membrane potential, DNA damage and/or proliferation. With the BD Accuri C6 Plus personal flow cytometer, you can perform dose-response studies right on your benchtop.
Dose-response studies can be time-consuming and labor-intensive, however, because of the sheer number of samples to process. A new BD product information sheet shows how the BD CSampler Plus automatic sampling accessory (Figure 1) can help. This option offers reliable, easy-to-use, walkaway convenience while adding minimal footprint to the BD Accuri C6 Plus.
Figure 1. BD CSampler Plus automatic sampling accessory
The BD CSampler Plus supports both 48- and 96-well plates (including 96-deep-well) and includes a 24-tube rack for standard 12 x 75-mm tubes. You can select up to three agitation cycles per well, as well as up to three wash cycles to minimize cross-well contamination. Data in the paper shows that the BD CSampler Plus will keep your cells in suspension over the course of your dose-response experiment, with consistent cell counts across runs of both a 24-tube rack and a 96-well plate.
Figure 2 shows a typical dose-response experiment analyzing the effects of nocodazole administration on the cell cycle in MDA-MB-468 cancer cells (one of two types analyzed in the paper).1 Nocodazole is an antineoplastic agent that causes cells to arrest in the G2 or M phase of the cell cycle. Cell cycle was assessed by staining with the DNA dye propidium iodide (PI) and the thymidine analog ethynyl-deoxyuridine (EdU), which is incorporated into newly synthesized DNA during the S phase. An antibody against phosphohistone H3 (pHH3) was added to differentiate M-phase cells from cells in other phases.
The dot plots show cell cycle analyses for untreated cells as well as a representative sample of cells treated with 100 nM of nocodazole. Treated cells showed more than a 10-fold increase in M-phase cells and more than a 3-fold increase in G2-phase cells, with corresponding decreases in other cell cycle phases. The graph shows dose-response cell cycle curves for both breast-cancer cell lines, pinpointing the dosage at which M- and G2-phase arrest occurred. The reduction in M-phase cells at higher dosages can be attributed to cells arresting earlier in G2, thus never progressing to M, and to loss due to cell death.
With the BD CSampler Plus, you can culture, prepare, run and analyze samples in the same plate, without transferring them to individual tubes. Besides streamlining experimental workflow, automation offers walkaway convenience at runtime, allowing you to focus on science instead of experiment management.
Easy to use, simple to maintain and affordable, the BD Accuri C6 Plus personal flow cytometer is equipped with a blue laser, a red laser, two light-scatter detectors and four fluorescence detectors. A compact and transportable design, fixed laser alignment, pre-optimized detector settings and automated instrument QC result in a system that is simple to use.
Tips & Tricks
BD Accuri™ resources
Last month, we highlighted the BD Accuri™ white papers available on our Resources page. But that’s not all. You’ll also find technical bulletins, experiment sheets, product information sheets, software templates, documentation and much more. Topics include kits and templates for popular application areas such as human and mouse T cells, cell cycle and DNA, and stem cell research; guides to absolute counting, apoptosis methods and identifying human blood populations; and tools such as a compensation calculator and more.
This section highlights interesting recent articles that describe research using BD Accuri flow cytometers.
Cancer therapy development
Sierra Rivera CA, Franco-Molina MA, Mendoza-Gamboa E, et al. Effect of bovine dialyzable leukocyte extract on induction of cell differentiation and death in K562 human chronic myelogenous leukemia cells. Oncol Lett. 2016;12:4449-4460. PubMed
Stem cell expansion
Morhayim J, van de Peppel J, Braakman E, et al. Osteoblasts secrete miRNA-containing extracellular vesicles that enhance expansion of human umbilical cord blood cells. Sci Rep. 2016;6:32034. PubMed
Safi C, Olivieri G, Campos RP, et al. Biorefinery of microalgal soluble proteins by sequential processing and membrane filtration. Bioresour Technol. 2017;225:151-158. PubMed
Longin C, Degueurce C, Julliat F, Guilloux-Benatier M, Rousseaux S, Alexandre H. Efficiency of population-dependent sulfite against Brettanomyces bruxellensis in red wine. Food Res Int. 2016;89:620-630. PubMed
Class 1 Laser Product.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
1 All reagents and kits are compatible with both the BD Accuri C6 Plus and the BD Accuri™ C6 flow cytometer systems. Data was generated on the BD Accuri C6 Plus. Information about BD reagent kits, BD Accuri™ C6 and BD Accuri C6 Plus software templates, and BD CSampler™ and BD CSampler Plus automation options is available at www.bdbiosciences.com/accuri.