(96-well w/ Stoppers)
|Oris™ Pro Plates|
(96-well w/ BCG)
|Oris™ Pro 384 Plates|
(384-well w/ BCG)
|Diameter of Well – Bottom||6.3 mm||6.58 mm||3.3 mm|
|Diameter of Well – Top||6.45 mm||6.96 mm||3.7 mm|
|Diameter of Cell Free Detection Zone||2 mm||2 mm||2 mm|
|Well Volume||400 µL||392 μL||131 µL|
|Suggested culture medium per Well||100 µL||100 µL||20 µL|
|Area of Outer Annular Region (cell seeding region)||28.03 mm2||30.86 mm2||6.86 mm2|
|Area of Central Detection Zone||3.14 mm2||3.14 mm2||3.14 mm2|
|Plate Height||14.85 mm||14.4 mm||14.4 mm|
|Plate Height with Lid||17.9 mm||17 mm||16.5 mm|
|Offset of Wells (A-1 location, X)||14.32 mm||14.38 mm||12.13 mm|
|Offset of Wells (A-1 location, Y)||11.25 mm||11.24 mm||8.99 mm|
|Distance between Wells||9.0 mm||9.0 mm||4.5 mm|
|Well Depth||12.1 mm||10.9 mm||11.5 mm|
|Thickness of Well Bottom||0.25 mm||190 μm ±10%||190 μm ±10%|
|Storage||4°C||15 – 30°C||15 – 30°C|
If your imaging system is provided with a software package, your vendor can help you with Oris™ quantification, and may already have a routine specifically designed for Oris™. See this application note for an example.
Some plate readers have limited clearance and may not accept the plate with the lid on.
The mask provided with the Oris™ assays has an aperture diameter of 2.1 mm while the size of the Detection Zone formed by the stopper is 2.0 mm. The larger mask aperture is designed to allow some fluorescent background signal to included in the light path in order to bring the detector into the appropriate dynamic range, as illustrated in this technical note.
Cells are typically visible under bright field optics without staining, especially if phase contrast optics are available. However, staining makes cells easier to see, and can reveal physiological states that bright field optics cannot. The Oris™ and Oris™ Pro Assays impose no restrictions on choice of stain, and you can use multiple stains simultaneously if desired.
To quantify migration/invasion via area closure, we recommend a fluorescent cytoplasmic stain such as TRITC-phalloidin in order to maximize signals for your detector.
To quantify migration/invasion by counting cells, we recommend a nuclear stain such as DAPI. Restricting the stain to the nuclei creates smaller objects for imaging than do cytoplasmic stains, providing greater separation between individual cells and therefore more accurate counts.
You may also consider pre-labelling cells prior to seeding in the Oris™ assays. However, some stains may impact migration and/or invasion, potentially creating experimental artefacts.