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  • Product Name: Multi-layer Flasks
  • Material: PS( Multi-Layer Flask) HDPE(cap)
  • Sterilization:

● 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







5-layer cell culture flask,straight neck, plug seal cap,TC treated




5-layer cell culture flask,straight neck,vent cap,TC treated




Culturing Technique 



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 .)
TipsA 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 de¬tached cell suspension into a NEST conical tube. 13. Rinse with additional wash buffer as needed.


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