TUNAIR™ polypropylene Erlenmeyer flask Kit, 300ml, No-Baffle

Additional Key Details:

• Comes complete with twelve (12) polypropylene 300 ml No-Baffle TUNAIR™ Shake Flasks, twelve (12) two-piece caps, and fifteen (15) dri-gauze filters.

Additional Description:

The Dri-Gauze filter is 0.22 micron and constructed of nitrocellulose. Each filter can be used 10 - 12 times. The Dri-Gauze filters can be autoclaved in the cap and left in place until a new filter is needed.

The two piece cap assemblies are constructed of polypropylene and are resistant to most solvents. All flasks and caps are fully autoclavable prior to reuse and filter linings can also be autoclaved or simply replaced.

Replacement Instructions:

To replace the filter lining in the cap assembly, simply pinch the flanges of the inner-closure shell until they snap loose. Then, pull apart and remove used lining. Replace the lining by sandwiching it between two parts of the cap and snap the cap back together. When reassembling the cap, ensure the flanges from the inner piece snap into the mated grooves in the outer piece. This will ensure the cap assembly stays together during use. All TUNAIR™ flasks and caps can be cleaned by soaking in water with a light detergent solution to loosen dirt and contaminants, then air dry.

Flask Dimensions:

Flask Size: 300 ml
Working Volume: 100 ml
Base Diameter: 3.25” [8.25 cm]
Neck Diameter: 1.75” [4.45 cm]
Height: 6.00” [15.24 cm]
Weight: 0.01 lbs. [0.004 Kg]

Mixing:

No-Baffle (0 Baffles): Normal Throws

Shaker Speed:

1” Throws: 300-400 rpm or possibly higher
2” Throws: 150-200 rpm or possibly higher

Material:

All TUNAIR flasks and caps are constructed of chemical resistant polypropylene. All flasks and caps are fully autoclavable.

300 ml flask:

Allows for approx. 150 ml (max) of working volume for culturing cells

2.5 Liter flask:

Allows for approx. 1 liter (max) of working volume for culturing cells

No-Baffle: 

The bottom inside of the flask is smooth and flat. This configuration will generate a single vortex within the media inside the flask when fastened to an orbital shaker. The shaker will need to operate at speeds above 160 rpm in order to form that vortex in the media.

Two Piece Cap:

The 300 ml flask and 2.5 L flask both have a special cap and filter system that promotes gas exchange and maintains sterility of the growth culture during use. The inner portion of the cap unsnaps from the outer portion by pushing in the tabs on the side of the cap assembly and then pulling the insert out.

0.2-micron filters: 

There are two different types of 0.2-micron filters available for the TUNAIR flasks. These filters allow for maximum gas exchange while maintaining sterility.

Dri-Gauze – This is a paper filter than can be used approx. 6 to 8 times before having to be replaced. This filter can be autoclaved while in the cap, on the flask, for 6 to 8 times.

Silicone – This is a silicone filter that will last the life of the flask. This filter can be autoclaved while in the cap and on the flask for many times.

Cap & Filter assembly: 

Once you have the 2-piece cap assembly apart (either the 300 ml cap or the 2.5 L cap) place the corresponding filter size inside the cap and the re-assemble the inner and outer cap pieces making certain to align the tabs with their associated holes. Once clipped into place, make certain the filter completely covers the cap assembly openings. 

Attaching the flasks to your shaker: 

IBI TUNAIR flasks can be used with nearly all standard shakers and shaker clamps. The 2.5-liter flask typically fits best into the 2-liter shaker clamp. The 300 ml flask typically fits best in the 250 ml shaker clamp.

Preparing and Cleaning the TUNAIR flask: 

Your TUNAIR flasks are constructed of polypropylene plastic which allows them to be autoclaved. Once you have prepared your media inside the flask you can place the cap & filter on the flask and insert into the autoclave. Set the autoclave for wet cycle.   

Once you have harvested your cells, wash the TUNAIR flask in a mild dish soap such as Dove Dish Soap then rinse well with DI water. Do NOT rinse the Dri-Gauze filter between uses. The silicone filter can also be rinsed with DI water if needed. Observe the condition of the filters between runs to make certain there are no holes or tears in the filter. If there are, replace the filter immediately.

Cell Growth Evaluation of Commonly Used Shake Flasks:

TUNAIR™ flasks were compared to conventional flasks using four different types of microorganisms; Escherichia coli, Saccharomyces cerevisiae, Penicillium avellaneum, and Streptomyces chartreusis. The aeration capacities of the shake flasks were determined by the sulfate oxidation method, and the values shown below are presented as oxygen absorption rate (OAR) in mM oxygen/L/Min. The growth rates of E.coli and S.cerevisiae were expressed as optical densities (OD) at 555 mM. For S.chartreusis and P.avellaneum growth rates were evaluated by percent sedimentation. For E.coli and S.cerevisiae, the growth rates were determined after an 18-hour incubation period; for S.charteusis, a 24-hour incubation period; and for P.avellaneum, a 72-hour incubation period. Growth and OAR evaluations were carried out with 3-9 replicates and statistically analyzed using Turkey’s w-procedure. See results below.

Growth Chart:

Growth Morphology:

M, mycelial; P, pellet; MP, mixed mycelial. The mycelial growths mostly adhered to the walls of the flask, which accounted for the low overall sedimentation value.

Growth Comparison of Saccharomyces Cerevisiae in TUNAIR™ Shake Flasks and Brand C Shake Flasks:

The experiment was done on a New Brunswick INNOVA 44 shaker incubator.
It was conducted at different speeds – 200 rpm & 300 rpm.
Strain: Saccharomyces Cerevisiae
Medium: YPD broth (Yeast Extract Peptone Dextrose)
Flasks: IBI TUNAIR™ 300 ml Flask and Brand C 250 ml Growth Flasks
Cell Analyzer: Vi-Cell XR

Growth chart: S.Cerevisiae Growth Chart

Experiment Summary:

In this experiment, standard volume--60 & 50 ml YPD--was used for TUNAIR™ and Brand C flasks, which is 20% capacity of the flasks. The flasks were incubated at 30°C at speeds of 200 rpm and 300 rpm for 28 hours. After taking viable cell counts, it was found that under low speed (200 rpm) both TUNAIR™ and Brand C flasks contained yeast cultures of higher cell densities when compared to the higher speed (300 rpm) cell culture flasks. It was also noted that the TUNAIR™ flask had higher cell density at 200 rpm and 300 rpm when compared to the Brand C flask. Although these data indicate TUNAIR™ flasks support higher density cell growth, we expected that higher speed (rpm) should produce higher cell density due to the higher dissolved oxygen concentration and better dispersion of cells. This unexpected data might be due to frozen cell culture stock, which might take a longer time to adapt to the environment.

More experiments are required and are being carried out at this time. The focus of these experiments will involve:
• Varying shaker speeds and varying volumes of medium in the TUNAIR™ and Brand C flasks
• Comparative studies between TUNAIR™ 2.5-L growth flasks and Brand C 2-L flasks
• Varying shaker speeds and varying volumes of medium in the larger TUNAIR (2.5 L) and larger Brand C (2 L) flasks

Reference Papers:

1. Method to Increase the Yield of Eukaryotic Membrane Protein Expression inSaccharomyces Cerevisiae

2. Optimisation of Recombinant Production of Active Human Cardiac SERCA2a ATPase