Falcon® Cell Culture Multi-Flask
An easier, more productive way to culture cells
Falcon Cell Culture Multi-Flasks enable you to grow more cells
faster and easier, thereby making your cell culture workflow
more productive.
The tissue culture-treated Falcon Multi-Flasks are available
in 3- or 5-layer formats that provide 525 cm2 or 875 cm2 cell
growth surface area. They can be used with a wide range of
liquid volumes (up to 50 mL/layer).
Features
• Even distribution of media across all layers for
homogeneous cell populations.
• Ability to mix in the Falcon Multi-Flask saves time and
reduces risk of contamination.
• Flexible design lets you pour or aspirate/recover cells
using a pipet.
• Consistent surface treatments for predictable scale-up.
• Manufactured in compliance to cGMP standards.
Designed to Fit Your Protocol
Simplify your scale-up. Falcon Multi-Flasks, offer the same
footprint, the same reagent volumes, and the same cell seeding
densities as 175 cm² flasks. Plus, you’ll be using the same
proven surface treatment as all other Falcon flasks.
Improves Your Cell Culture Productivity
Falcon Multi-Flasks deliver a thoughtful design that
simplifies your workflow. You can eliminate multiple
steps and reduce the risk of contamination.
• Pipet access allows you to aspirate media to exchange
with fresh media and recover cells without pouring.
• The mixing port allows rapid mixing within the Falcon
Multi-Flask to create your cell suspension, transfect your
cells, or add other reagents directly to the flask. The port
also provides for a uniform distribution of media and cells
to facilitate homogeneous cell growth on all layers.
More Consistent Cell Growth
Falcon Multi-Flasks’ even distribution of media, proven
vacuum-gas tissue culture surface treatment, and
effective gas exchange all combine to provide an
optimal cell culture environment. The result is higher
cell yield and a more homogeneous cell population.
Visit www.corning.com/lifesciences for more information.
FALCON
®
PREDICTABLE SCALE-UP
14
5
4
5
10
Cell Number x 10
Cell Number x 10
5
12
8
6
4
2
3
2
1
0
Corning T175 Flask
(1-layer)
Falcon Multi-Flask
(3-layer)
Falcon Multi-Flask
(5-layer)
0
BHK-21
CELL CULTURE VESSEL
Expected Yield
LnCap
Hep-G2
EcoPack 2-293
CELL LINE
Actual Yield
T-175
Falcon Multi-Flask (3-layer)
Falcon Multi-Flask (5-layer)
Three and five times more BHK-21 cells were grown and recovered from 3- and 5-layer Falcon® Multi-Flasks than T-175 flasks (left). Cell yields per
cm2 were equivalent in 3- and 5-layer Falcon Multi-Flasks and T-175 flasks for BHK-21, LnCap, Hep-G2, and EcoPack™2-293 cells (right).
CONSISTENT CELL PHYSIOLOGY
5.0
Calcium Mobilization
Fold Increase Over Control
(mean + SD)
CHO-M1 cells were grown in a T-175 and a 3-layer Falcon
Multi-Flask for 72 hours. Cells were harvested and re-seeded
on 96-well PDL plates for GPCR Calcium mobilization assay.
Cells were challenged with M1 agonist, Carbachol (300 µM)
and responses were recorded 1 minute following compound
addition. Data is expressed as fold-increase in calcium flux
compared to control cells challenged with assay buffer
(no drug). No significant difference in agonist response
was observed between cells harvested from either vessel
type. Data shown is representative of three independent
experiments.
4.0
3.0
2.0
1.0
0.0
T-175
Falcon 3-layer
CELL CULTURE VESSEL
SIMPLIFIED WORKFLOW
• Pipet access eliminates the need for pouring. It also
facilitates the use of a wide range of liquid volumes
(up to 50 mL/layer), efficient recovery of valuable cells
and reagents, and minimizes the risk of contamination.
• Mixing port eliminates mixing outside the vessel, saving
you time and effort. This unique in-vessel mixing allows
for rapid and uniform distribution of cells and reagents.
• Compatibility with similar volumes of reagent usage
and cell seeding densities as T-175 flasks makes scaling
up a familiar and simple process.
CONSISTENT CELL GROWTH
This figure illustrates
homogeneous cell growth
between layers of Falcon®
Multi-Flasks. BHK-21 cells
grown to >80% confluence in
3-layer Falcon Multi-Flasks and
control T-175 flasks were fixed
and stained with crystal violet.
Control flasks and individual
layers of the multi-flask vessels
were cut and scanned.
Top
Middle
IDEAL FOR PLURIPOTENT STEM CELLS
Human Mesenchymal Stem Cells were cultured in MSCGM™ medium
(Lonza) and seeded at 4,000 to 5,000 cells/cm2 in a T-175 Falcon Flask and
a 3-layer Falcon Multi-Flask. The cells were cultured until they reached
80% confluence (~6 days). Cells were harvested, counted, and cumulative
population doublings of hMSCs from each vessel type were calculated over
five consecutive passages. Immunophenotypic properties of hMSCs grown
in the Falcon Multi-Flask as determined by flow cytometric analysis (FACS)
confirmed that hMSCs were negative for surface markers CD14, CD34,
CD45, CD79a, and HLA-DR and positive for expression of surface markers
CD73, CD90, and CD105. Respective isotype controls for each marker
antibody are shown in grey.
COMPATIBLE WITH YOUR CELL LINES
Diverse cell lines and primary cultures (with and
without serum) can be grown and efficiently recovered from Falcon Multi-Flasks. Expected yield was
determined using average cell yield from control
T-175 flasks multiplied by three and five times for
the 3- and 5-layer Falcon Multi-Flasks, respectively.
Bottom
Corning® T-175 (control)
Falcon® Cell Culture Multi-Flask
An easier, more productive way to culture cells
PRODUCT SPECIFICATIONS
WORKING VOLUME RANGE
>5 mL per layer for dissociating
ORDER INFORMATION
Falcon Cell Culture Multi-Flask
<50 mL per layer for cell expansion
Description
MOLDED-IN GRADUATIONS
0 to 50 mL per layer in 10 mL increments
GRADUATION ACCURACY 10%
CAP VENT MEMBRANE
0.2 μm hydrophobic membrane
CELL GROWTH SURFACE
Tissue culture-treated, optically clear
Qty/Pack
Qty/Case
Cat. No.
2
3-LAYER TISSUE CULTURE-TREATED, 525 CM
2
12
353143
5-LAYER TISSUE CULTURE-TREATED, 875 CM2
1
8
353144
To place an order in Europe, contact Customer Service at:
tel: + 31 0 20 659 60 51, fax: + 31 0 20 659 76 73; email: [email protected]
For technical assistance, contact Technical Support at:
email: [email protected]
ACCESSORIES
Description
Qty/Pack
Qty/Case
Cat. No.
ASPIRATING PIPET 2 ML INDIVIDUALLY WRAPPED
50
200
357558
10 ML INDIVIDUALLY WRAPPED PAPER-PLASTIC
50
200
357551
10 ML INDIVIDUALLY WRAPPED PLASTIC-PLASTIC
50
200
356551
50 ML POLYPROPYLENE CONICAL TUBE
25
500
352070
175 ML POLYPROPYLENE CONICAL TUBE
8
48
352076
225 ML POLYPROPYLENE CONICAL TUBE
8
48
352075
Falcon Pipets
Falcon Tubes
Corning Incorporated
Life Sciences
Corning acquired the Discovery Labware Business including the Falcon® brand.
For information, visit www.corning.com/discoverylabware.
For a listing of trademarks, visit us at www.corning.com/lifesciences/trademarks.
All other trademarks are property of their respective owners.
Corning Incorporated, One Riverfront Plaza, Corning, NY 14831-0001
Corning BV
Fogostraat 12
1060 LJ Amsterdam
The Netherlands
Phone: +31 0 20 659 60 51
Fax: +31 20 659 76 73
[email protected]
www.corning.com/lifesciences
© 2012, 2013 Corning Incorporated
Printed in Europe 02/13
CLS-DL-CC-024 A4
U.S. Orders
Contact your authorized distributor to place your order.
Falcon® Multi-Flask
Frequently Asked Questions
FALCON
®
Why does media spill over to the other layers when moving the
Falcon Multi-Flask in and out of the incubator?
Transport the flask according to the user guide (see step 8) using the partition position
and keeping fluid away from the mix port. Do not carry Falcon Multi-Flasks in a
horizontal position as this may cause spillover of media between layers.
Why does media distribution become uneven when I place the
Falcon Multi-Flask in the incubator?
Fluid may migrate from one layer to the other. To prevent this from occurring, one
should:
• Ensure the incubator shelf is level.
• Position flasks across shelf without blocking the vented neck region.
• Ensure weight is distributed evenly throughout the shelf and the shelf can withstand
the weight of flasks with media.
• Use the opposite corner of the mix port to pivot the flask and lay it horizontally.
Ensure media does not flow back into the mix port or over the shelves into the
neck region.
I followed the user guide and set the Falcon Multi-Flask into the
incubator. A few hours later, I see uneven media distribution by layer.
What do I do?
• Repeat re-equilibration and partitioning steps (user guide steps 7 and 8).
• Ensure the work surface is flat while equilibrating and partitioning fluid in the
Falcon Multi-Flask.
• Inspect the flask at least 2 hours after seeding. Re-equilibrate and partition again,
if necessary.
• Minimize vibration on, or near, the surface where Falcon Multi-Flasks are housed.
What volume of media should I use?
Begin with the same media volume and cell seeding density on a per unit surface area
basis as your existing device. The surface area of the flasks are 525 cm2 for 3-layer
and 875 cm2 for 5-layer flasks. The maximum amount of media used should not
exceed 50 mL per layer.
What volume of dissociation reagent should I use?
We recommend ≥5 mL per layer of Falcon Multi-Flask.
Can I use the same volume of reagents as T-175 flasks?
No, the amount of media per unit area should be the same—not the actual total
volume. If you currently use a T-175 flask, simply multiply the volume by 3 or 5 times
depending on the Falcon Multi-Flask format used.
Why do I observe excessive frothing or bubbling of media and can this
affect my cell culture?
Over mixing can result in excessive frothing or bubbling of media, which can lead to
fluid migration from one layer to another. To avoid this, follow user guide step 5. While
the flask is in the mixing position, allow the media to drain only from the layer on top;
immediately tilt the flask in the opposite direction; and repeat. Ensure the flask is brought
back to the mix position (user guide step 4) before equilibration of media. Improper
mixing or pouring of media into the flask can also create foaming or bubbles. The correct
way to add media is to follow step 1 in the user guide and pour or pipet gently along the
slope of the flask lid.
How can the flasks be stacked in an incubator?
Ensure the flasks lock into position using the stacking ribs. Position the stacks of flasks
evenly in the incubator to prevent the shelves from bowing. Allow adequate room for air
circulation through the vented cap of the flasks. When stacking the Falcon® Multi-Flask
on incubator shelves that are above shoulder height, ensure the media distribution by
layer remains equal. The use a step-stool can help facilitate flask placement.
What is the best way to add cells to the vessel?
Add the desired volume of growth media to the flask and keep it in an upright position.
Add cell suspension stock (at least 10% final volume) directly into the growth medium
using a 10 mL pipet.
Is pre-wetting of the flask or pre-warming of the media in the flask
required?
Neither is required, nor are they recommended unless you are applying a coating to the
flask growth surface.
The 5-layer Falcon Multi-Flask will not fit under my microscope.
What do you recommend?
With some inverted microscopes, the condenser can be removed or moved to
accommodate the 5-layer flasks. If this is not possible, a control flask (T-175) can be run
side-by-side to provide an indication of cell morphology and confluence.
Why is cell growth patchy when I view the Falcon Multi-Flask under the
microscope?
Uneven cell/media distribution most likely occurs from inadequate mixing. Refer to steps
4 and 5 of the user guide for the proper procedure. Don't use a clumpy cell suspension
stock. Ensure cells are adequately triturated to generate a single cell suspension.
Why am I getting media in the neck of the flask?
Media should not enter the neck of the flask if the Falcon Multi-Flask is used properly.
Check to ensure the flask is not upside down. The logo should face up when the flask is
laid horizontally for incubation. Over-rotation when mixing cells in the vessel can result
in media getting into the neck area of the flask. To avoid this, please follow steps 4 to 6 in
the user guide.
Why are my cells not attaching?
If your cells normally attach to a single layer flask, then they will attach to the surface of
the Falcon Multi-Flask. Ensure the flask is laid in the correct position with the logo facing
up. In this position, cells will adhere to the tissue culture surface on the base of each layer.
Can I pour my cell suspension into the flask?
Yes, however we recommend that to avoid foaming of media, use a 50 or 100 mL pipet
(Cat. Nos. 357550 and 357600). Insert the pipet past the logo and stream liquid along
the top slope of the flask into the first layer.
Can I pour my cells and reagents out of the flask?
Yes, you can pour or use a pipet to recover the cells during harvesting. If cells clump,
mechanically dissociate the cells by titrating the suspension.
How can I maximize the recovery of cells and reagents?
• To minimize residual volumes, pour as per step 13 in the user guide. Allow liquid to
pool on the mix port side prior to pouring with the logo down.
• For maximal recovery of cells and reagents, follow steps 11 and 12 in the user guide.
First, aspirate by positioning the pipet tip towards the mix port region. Then, pool
liquid into the top layer and tilt the flask to partition liquid away from mix port.
• For high density cell cultures, additional rinses are recommended. Visually inspect the
base layer to determine how many rinses are required.
Why am I not getting the expected cell yield?
• You may not be recovering all the cells. Additional rinses may be required, as
noted above.
• You may have uneven distribution of cells and media. See the user guide for proper
mixing, equalization and partitioning procedures.
• You should avoid lengthy delays between cell seeding and incubation so that cells do
not settle to the bottom of the vessel.