5-Layer Cell Culture Flask

Product#: 73100


  • Seal
  • Vent
Ships in 24 hours

5-Layer Cell Culture Flask

Cat. No. See below

Nest Multi-Layer Flasks provide 870 cm2 cell growth surface area. The tissue culture treated NEST Multi-layer Flasks enable you to grow more cells faster and easier, thereby making your cell culture workflow more productive, easier, and in a more convenient manner.
Product features
  • Vacuum plasma TC treatment for the best cell attachment
  • Sterilized by E-Beam
  • Non-Pyrogenic, DNase/RNase FREE
  • Both plug seal cap and vent cap are available
  • Growth area: 870cm2
  • TC treatment for better cell attachment
  • Sterilized by E-Beam, non-pyrogenic
  • Tips: To avoid foams and bubbles causing the media in upper layer flowing into the bottom layer, users should be gently when operating the multi-layer flasks.
  Product ID Cell Growth area (cm2) Cap Style Recommended Medium Volume (mL) Treat Packing Unit 
73100_Seal-diagnocine 73100_Seal 870 Sealed Cap 100-150 Yes 1/pk, 8/cs
2-731002-1.jpg 73100_Vent 870 Vent Cap 100-150 Yes 1/pk, 8/cs
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 up right 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.

Media Removal:
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.

Cell Harvesting:

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.

  • Cell biology research
  • Cytology research in animal and plant and basic medical school
  • Small-scale cell culture, cell screening
  • Cell threapy direction research
  • Stem cell research
  • Tissue culture
  • Biochemistry, medicine, microbiology
  • Genetic engineering, antibody engineering drugs, vaccines, etc
  • Microplate chromogenic reaction plate use
Working Volume Range:   >5mL per layer for dissociating, >30mL per layer for cell expansion
Molded-In Graduations:    0 to 50mL per layer in 10mL increments
Graduation Accuracy:    10%
Cap Vent Membrane:   0.22um hydrophobic membrane
Cell Growth Surface:   tissue culture-treated, optically clear

  • Such products are provided with Treated, Non-Treated products, applicable for adherent cells or suspension cell culture
Quick Link to Treated Cell Culture Flask treated.png
Quick Link to Non-Treated Cell Culture Flask Non-Treated.png
Quick Link to 5-Layer Cell Culture Flask 5-layer.jpg


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