Features:
- High clarity medical grade polystyrene material
- High performance with 5 growth surfaces
- Growth area: 870cm²
- TC treatment for better cell attachment
- Sterilized by E-Beam, non-pyrogenic
- Vent cap
Adding Media and Preparing Cell Suspension:
- Mix cell suspension with medium: Prepare cell suspension of required concentration in a container and mix. Recommended volume is about 30- 50 mL per layer.
- Add the mixed liquid into the Multi-layer Flask slowly with serological pipettes. To avoid foams and bubbles, allow liquid stream to flow along the slope of the Multi-layer Flask. (Save a little liquid in pipette for each dosing)
Tip: A 10 mL pipette allows media to be dispensed at the bottom of the vessel. A 25 mL pipette allows media to be dispensed just past the NEST Logo.
- Hold the Multi-layer Flask upright with the Logo facing you and tilt clockwise to a 45° angle on a flat work surface to partition the liquid into each layer.
- While holding the Multi-layer Flask at a 45° angle, gently lay it flat onto the work surface with logo facing up.
- After placing the Multi-layer Flask flat on a work surface, gently rock back and forth and side-to-side to distribute cells evenly onto culture surfaces.
- Repeat Step 3 to put the flask quickly and slightly into the incubator. Then, lay it flat as shown in Step 4 .
Media Removal
- Aspirating method: To aspirate or remove media, tilt Multi-layer Flask, with the NEST Logo facing you, counter-clock wise to a 45° angle while inverting the Multi-Flask toward you. Then, tilt Multi-layer Flask to the right, continuing to aspirate all residual media.
- Pouring method: With Logo facing you, pour spent media from Multi-layer Flask
Cell Harvesting
- Wash with buffer once and add dissociating reagent (≥5 mL per layer). Then, follow Steps 3-4 to distribute to dissociating reagent to each layer.
- Neutralize with inactivating solution and mix following Steps 3-4. Gently swirl to dislodge cells completely.
- Follow “Aspirating Method” protocol and collect cell suspension using a NEST 10mL serological pipet.
- Follow “Pouring Method”. Pour detached cell suspension into a NEST conical tube. 13. Rinse with additional wash buffer as needed.
