|Evaluation of MultiScreen-MIC Plates in Chemotaxis Assays|
|Evaluation of MultiScreen-MIC Plates in Invasion and Angiogenesis Assays|
|MultiScreen-MIC Plate: 96-well system for cell-based functional screening assays including migration, invasion, and chemotaxis|
|Extracellular HIV-1 Nef increases migration of monocytes|
Michael H. Lehmanna, Stefan Walterc, Loyda Ylisastiguid, Frank Striebelc, Vladimir Ovode, Matthias Geyerf, Jean Claude Gluckmand and Volker Erflea
Experimental Cell Research, Volume 312, Issue 18 , 1 November 2006, Pages 3659-3668 2006
|High throughput in vivo screening for bone anabolic compounds with zebrafish|
Angeleen Fleming, Masahiko Sato, Paul Goldsmith
Journal of Biomolecular Screening 10(8); 2005, 823-831 2005
|Screening Assay for the Identification of Deoxyhypusine Synthase Inhibitors|
Marc-Nicola Somner, Dorian Bevec, et al.
Journal of Biomolecular Screening, 2004, 9(5):434-438 2004
|Cell Based Assays|
|A cell-based assay for screening the uridine 5'-diphosphate-glucuronosyltransferase 1A inhibitory potential of new chemical entities.|
Jaime Padros, Donald Chun, Liangfu Chen, Pierre Rigolli, Themis Flarakos and Malle Jurima-Romet; Analytical Biochemistry 320 (2): 310-312
Analytical Biochemistry 320 (2): 310-312 2003
|Functional characterization of Glu298Asp mutant human endothelial nitric oxide synthase purified from a yeast expression system.|
Regina Golser, Antonius C. F. Gorren, Bernd Mayer and Kurt Schmidt
Nitric Oxide 8 (1): 7-14 2003
|Cell Based Assays|
|What cell density should I use in my assay when using the MultiScreenTM-MIC plates?||Every cell type calls for a different cell density. Before you set up your actual assay, you must run standardization experiments testing a range of cell densities and assess which cell concentration gives you the optimal results. You can adjust the cell: pore ratio to optimize your signal to background values. Pore densities for the different pore sizes are listed below.
We have used invasive and non-invasive breast cancer cell lines (MDAMB231 and MCF7), epithelial cell line (HT1080) and fibroblast cell lines (NIH3T3) in our MIC plates. We typically use 50,000 cells/well.
|What volumes should I use in my assay when using the MultiScreen-MIC plates?||We typically use 50 µl in the top well and 150 µl in the bottom well.|
|How long should I run an assay with the MultiScreenTM-MIC plates?||Chemotaxis and transmigration assays are typically run for 2-6 hrs. Invasion assays are typically run for 24-72 hrs. Angiogenesis assays are typically run for 12-24 hrs. It is best to optimize assay time best suited to your cell line by assessing your assay results at different time periods.|
|Is the membrane in the MultiScreen-MIC plates and Millicell inserts the same? What is the difference between PC membrane and PCF membrane?||PC is polycarbonate with PVP (Poly-vinyl-pyrrolidine) and PCF is PVP free-polycarbonate. The membrane in the MultiScreenTM-MIC plate is PVP-free polycarbonate and is tissue culture treated. We have Millicells that are PC, PCF and non-polycarbonate type membranes.|
|What is the well depth and maximum volume capacity of a MultiScreen plate?||The well depth of a 96 well MultiScreen plate is 1.245 cm. The well depth for a 384 well MultiScreen plate is 1.2 cm. The maximum working volume of a 96 well plate is 300 ul. The maximum working volume for a 384 well plate is 100 ul.|
|Assay Terminology commonly used with reference to MultiScreen-MIC plates||Chemotaxis: Directed movement of cells in response to a concentration gradient.
Migration: Random movement of cells.
Invasion: The movement of cells across a barrier (e.g. extracellular matrix) in response to a concentration gradient.
Transmigration: The movement of cells across another cell barrier in response to a concentration gradient.
Angiogenesis: Tubule formation exhibited by HUVEC (Human Umbilical Vein Endothelial Cells) in response to cell stimulating factors.
Co-culture: When you have more than one cell type in your assay it is called co-culture. These assays are typically run to assess cell-cell interactions and factors secreted by one cell influencing the behavior of the other cell.
|What type of assays can I perform on the MultiScreen-MIC plates?||he MultiScreenTM-MIC plates can be used for a variety of assays such as migration, invasion, chemotaxis, angiogenesis, transendothelial migration and co-culture type of assays.|
|What is the pore density in the MultiScreenTM-MIC plates?||Pore size Nominal pore density per cm2 3 mm MIC 2 X 106 5 mm MIC 4 X 105 8 mm MIC 1 X 105|
|Should I pre-coat the wells when using the MultiScreenTM-MIC plates?||This depends on the cell line you are using. Some cells need coating and some don’t. Again a quick literature search should help you decide the correct coating substance for your cell line.
Typical coating substances are collagen and gelatin. Extracellular matrix is typically used for coating wells when performing invasion assays and angiogenesis assays.
|How do I coat the wells when using the MultiScreenTM-MIC plates?||Use 30-50 µl volumes of coating substance at desired concentration in a buffer compatible with coating substance. Spread the solution gently across the membrane using a pipette tip taking care not to damage the membrane. An alternative approach is to pre-wet the membrane with buffer solution, aspirating and then applying the coating solution. Care must be taken to ensure that there is no drying of membrane prior to coating.|
|MultiScreen - MIC Plate|
|MultiScreen FL Plate User Guide|