Multi-layer Flasks
? 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
|
Cat.No. |
Description |
Qty./Pack |
Qty./Case |
|
731001 |
5-layer cell culture flask,straight neck, plug seal cap,TC treated |
1 |
8 |
|
731002 |
5-layer cell culture flask,straight neck,vent cap,TC treated |
1 |
8 |
?Culturing Technique
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 .)
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 .
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,? 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
?? 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 10mL serological pipet .
12. Follow Step 8 ?Pouring Method?. Pour de?tached cell suspension into a conical tube.
13. Rinse with additional wash buffer as needed.