Nest 5 Layer Cell Culture Flask, Plug Seal Cap, Straight Neck
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NEST® tissue culture / cell culture multi-layer (5-Layer) flasks are high throughput TC flasks designed to
make it easier and faster to grow a large number of cells at once. Nest® 5-layer culture flasks come sterile
with TC treated surface with an optional vented cap or plug seal cap. Multilayer flasks have a straight neck
for easy cell retrieval and high clarity medical grade polystyrene construction. Nest™ is now among the top
most trusted brands in the market for your delicate cell culture or tissue culture assays. All Nest multi-layer
flasks are individually sealed, wrapped and sterile and there are 8 flasks per case.
- 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
- Both plug seal cap and vent cap are available
How to correctly use multi layer Cell Culture Flasks?
1. Prepare cell suspension, pour liquid into the 5-layer cell culture flask slowly. To avoid foams and bubbles, use a serological pipette. Recommended volume is about 150ml to 200ml.
2. Place the flask upright for a period to balance the liquid volume in every layer. 2a. For small amounts of solution, tilt the flask along the corner connecting line to quicken the balance.
3. Put the flask quickly and precisely into the incubator.
4. The liquid will be spread over 5 growth layers uniformly.
5. Multi-layer flasks can be emptied as a single layer flask. To harvest cells add 25-35ml Trypsin-EDTA .
Adding Media And Preparing Cell Suspension:
1. 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.
2. 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 .)
Tips：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.
3. 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.
4. While holding the Multi-layer Flask at a 45° angle, gently lay it flat onto the work surface with logo facing up.
5. 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.
Tips: Take care to avoid foaming of medium, and not to spill liquid from each layer.
6. Repeat Step 3 to put the flask quickly and slightly into the incubator . Then, lay it flat as shown in Step 4 .
You may choose to either aspirate or pour the media from Multi-layer Flask.
7. 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.
8. Pouring method :
With Logo facing you, pour spent media from Multi-layer Flask
Tips: Aspirate media using a NEST 2mL or 10mL aspirating pipette.
9. Wash with buffer for one time and add dissociating reagent (≥5 mL per layer). Then, follow Steps 3-4 to distribute to dissociating reagent to each layer.
10. Neutralize with inactivating solution and mix following Steps 3-4. Gently swirl to dislodge cells completely.
11. Follow Step 7 “Aspirating Method” protocol and collect cell suspension using a NEST 10mL serological pipet .
12. Follow Step 8 “Pouring Method”. Pour detached cell suspension into a NEST conical tube.
13. Rinse with additional wash buffer as needed.