Multi-layer Cell Culture Flask
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Multi-layer Cell Culture Flask

Nest Multi-layer cell culture flasks are a cost effective way to scale up your cell cultures while saving time and effort. You can't make your cells grow faster, but you can spend less time in the tissue culture suite. 

Such as 5-layer flasks allow you to handle the equivalent of 5 X 175cm² flasks at once. As an added bonus they also take up less space per unit growth area as compared to single 175cm² flasks. This means 5X as many cells can be grown in the same incubator.

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Introduction
• Made of high clarity, 100% virgin polystyrene.
• Sterilized by E-beam, SAL=10-6.
• Non-Pyrogenic, DNase/Rnase free.
• 3-Layer Cell Culture Flask Growth area: 520 cm². 5-Layer Cell Culture Flask Growth area: 870 cm².
• Individually packaged in sterile bag.

Use Guide：


1、Mix cell suspension with medium: Prepare cell suspension of required concentration in a container. Recommended volume is about 30- 50mL per layer.
2、Add the mixed liquid into the Multi-layer Flask slowly with serological pipettes. To avoid foam and bubbles, allow liquid stream to flow along the slope of the Multi-layer Flask. (Save a little
liquid in pipette each time.)
3、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.
4、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.
5、While holding the Multi-layer Flask at a 45° angle, gently lay it flat onto the work surface with logo facing up.
6、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.
7、Tips: Be careful to avoid foaming of medium, and not to spill liquid from each layer.
8、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.
9、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.
10、Pouring method: With Logo facing you, pour spent media from Multi-layer Flask
11、Tips: Aspirate media using a NEST 2 mL or 10mL aspirating pipette.
12、Wash with buffer for one time and add dissociating reagent (≥5mL per layer). Then, follow Steps 3-4 to distribute to dissociating reagent to each layer.
13、Neutralize with inactivating solution and mix following Steps 3-4. Gently swirl to dislodge cells completely.
14、Follow Step 7 “Aspirating Method” protocol and collect cell suspension using a NEST 10mL serological pipette.
15、Follow Step 8 “Pouring Method”. Pour the cell suspension into a NEST conical tube.
16、Rinse with additional wash buffer if needed.
17、Search "NEST Multi-layer Flask" video on NEST website or Youtube.