An OptiCell has the same footprint of a microtiter plate. The frame is less than 6 mm (1/4 in) thick. The thickness of the growth chamber is 2 mm.

The recommended volume for cell/media is 10 ml.

One OptiCell membrane provides 50cm² surface area for cells to attach. Users have the option of using both membranes in one OptiCell, which provide 100cm² surface area. The two membranes are identical.

The membrane is a polystyrene matrix material. It is 75 µm thick. It is permeable to O2, CO2, and N2 gas.

Yes. The surface of the OptiCell membrane is Corona treated to allow for cell adhesion. Manufacturers of tissue culture flasks and plates use Corona treatment to make the surface hydrophilic.

No. However, we can do a special production run whereby there is no treatment added to the surface. Please contact us for more information.

The OptiCell is resistant to temperature between -196°C to 70°C. It is fragile below -152°C temperature.

The water evaporation rate is 0.016 ml/day in a 37°C incubator with 90% relative humidity.

Use an alcohol pad to push down the port back into its slot.

Currently there is only one size available.

No. The OptiCell tip will not pierce human skin at less than or equal to 200 grams of force. The blunt, polished stainless steel cannula tip protects the integrity of the self-sealing media cap septa and OptiCell ports.

No. Regular needles will damage OptiCell ports and membrane, and should not be used with OptiCell.

Each OptiCell port self-seals after up to 20 penetrations with OptiCell tips. Care should be taken to minimize the number of insertions and to direct insertions carefully toward the center of the port to reduce the risk of damage, contamination, and leaks.

Yes, we have tested P-1000, P-200, and P-20 tips for inserting volumes of less than 1 ml.

Each rack holds 20 OptiCell that provide the same surface area as roughly 26 T75 flasks.

No.

The film is only gas permeable. This is a molecular property of the polystyrene film used in the OptiCell. The film allows the transfer of O2, CO2, and N2 gas into and out of the OptiCell. Bacteria, viruses, and mycoplasma cannot transfer through the film.

We do not recommend re-using OptiCell.

Like any cell culture device, the growth rate of the cells in OptiCell varies depend on the individual cell line. Majority of the cell lines duplicate every 20-24 hours. Some can be slower or faster. For example, HUVEC cells duplicate every 36 hours whereas CHO-K1 cells duplicate every 12-16 hours.

A variety of different cell lines have been tested in OptiCell. These include:
human breast carcinoma - T47D
adenovirus transformed human kidney epithelial cell line - HEK-293
normal mouse fibroblast cell lines - 3T3 and BLK CL.4
normal human umbilical endothelial cell line – HUVEC
mouse melanoma cell line - B16
mouse macrophage cell line - IC-21
human cervix adenocarcinoma line – Hela
Chinese hamster ovary epithelial line - CHO-K1
mouse lung carcinoma line - LLC1
human T lymphoblast – Jurkat
human B cell lymphoma – Raji
mouse hybridoma cell lines - B72.3 and 9D9

OptiCell is an ideal tool for maintaining cell lines. It saves incubation space by five times as compare to that of T75 flasks. The amount of medium needed for cell culture in OptiCell is 10 ml for 100cm² growth area. It also provides a self-sealed compartment ideal for virus-related cell culture. Most importantly, OptiCell greatly facilitates the subsequent setups of assays (e.g. cytostaining, magnetic separation, antibody collection from hybridomas, transfection, etc.). For example, if you would like to perform immunocytostaining of the cells, you can fix the cells in OptiCell with 100% methanol, and then either directly stain the cells in situ or cut the films off from the frame for smaller scale staining. The leftover films can be stored for multiple repeats without having to set up the cell culture again.

Yes. Besides mammalian cells, OptiCell have been used to culture insect cells, virus, bacteria, nematode, and yeast.

Both adherent cells and non-adherent cells can be cultured in the OptiCell.

You can trypsinize the cells in OptiCell (Please see the protocol under applications). If you don’t prefer to enzymatically detach the cells, instead of scraping the cells off, you can perform trypsin-free detachment in OptiCell using an ultrasonic water bath (Please see the protocol under applications). In both protocols, you can observe the cells under a microscope to make sure they are all off.

Seed cells in an OptiCell and incubate it at horizontal position. By gravity, the cells would fall and attach to the bottom membrane (first side) of the OptiCell. In order to allow the cells that haven’t attached to the first side of OptiCell to fall down to the second side, the OptiCell has to be flipped during the incubation. The optimal time that enables equal cell attachment on both surfaces of the OptiCell membranes is called Flipping Time (FT). FT enables the best usage of the entire 100cm² growth area for adherent cells. Here are some examples of FT for a few adherent cell types. Compare the speed of attachment of your cell culture to some of these cell lines will help you determine the best FT for your particular cell type.

CHO-K1 10 minutes
T47-D 15 minutes
3T3 20 minutes
HUVEC 30 minutes
BLK CL4 40 minutes
HEK293 40 minutes
IC-21 45 minutes
B16 55 minutes

OptiCell does not come pre-coated. However, users can easily coat the OptiCell growth surfaces by injecting collagen, fibronectin, etc. into OptiCell through one of two access ports and allowing the coating to cover the growth surface area. The excess can then be withdrawn through one of the ports. We recommend storing or drying the coated OptiCell in a vertical position.

The recommended minimum media volume is 7 ml for one OptiCell in order to avoid the two membranes from collapsing and touching.

The bubbles you observed are not cause by media evaporation but rather from the microscopic bobbles (tiny bubbles) left behind. It is OK to remove them with a syringe and a tip.

Yes. BioCrystal lab and OptiCell customers have successfully tested OptiCell for a number of primary cell types, including rat cortical neurons, rat adipocytes, human dermal fibroblasts, human primary endothelial tumor cells, mouse keratinocytes, mouse hepatocytes, mouse splenocytes, mouse dendritic cells, mouse embryonic stem cells, etc.

OptiCell provides a completely self-sealed sterile system for hybridoma growth. The concentration of the antibody collected from OptiCell is 2-3 times higher than that from flask. The total quantity is also higher. It allows harvest of antibody 3 times a week. Most conveniently, there is no need to spin down cells every time at collection of supernatant. One OptiCell lasts more than a month for continuous antibody production. Please check out the most updated protocol for hybridoma antibody production in OptiCell.

Insect cells do not strongly adhere to culture surfaces. Insect cell culture can readily be passed from a "full" OptiCell to another by a simple “re-suspension” of the cells by rapping the OptiCell on the palm of the hand. Pull out 2 ml of cell suspension with the syringe and transfer the cells to a pre-filled new OptiCell. Instead of warming up 500-1000 ml of media at a time for cell passage you can pre-fill several OptiCell and store them in the refrigerator. When you need to passage cells you pull out one cassette and warm up within minutes in a water bath. Please check out the most updated protocol for culturing insect cells in OptiCell.

Yes. You can freeze cells in freezing medium containing 5-10% DMSO in OptiCell at -80°C for up to a month or in a -152°C deep freezer permanently (protocol). When thawed and replaced with culture medium, a significant amount of cells will stay attached to the OptiCell membrane and resume growth immediately.

Wipe the OptiCell ports with a 70% alcohol pad before access the OptiCell.

Absolutely. We have specially designed an OptiCell mailer for this purpose (see the ordering information for OptiCell mailers). We highly recommend mailing live cell culture at their log phase of growth, delivering the cell culture within 4 days, and changing with fresh culture media upon arrival.

After use, dispose of OptiCell cultures and syringes according to standard biohazard waste disposal procedures. Dispose of OptiCell tips and OptiCell knives in approved labeled sharps containers.

Absolutely. Just remove culture media and inject 10 ml of fixative in the OptiCell. Other than acetone (damages OptiCell membrane), you can use most of the common fixatives on OptiCell. Please see the OptiCell Film Tolerances Sheet for detailed information.

First, fix the cells directly in OptiCell (OptiCell Film Tolerances Sheet). Then cut the membranes off from the frame along the edges using an OptiCell Knife. Use a pair of scissors to cut the membrane into small pieces and place them in a multi-well plate for smaller scale staining. After staining, the OptiCell membrane can serve as a cover slip to be mounted onto a regular slide for storage (see OptiCell Film Tolerances Sheet for the proper mounting material).

Yes. OptiCell membranes have low background fluorescence and will not interfere with fluorescent assays.

Yes, you can use certain brand of immersion oil on OptiCell membrane. Please check the OptiCell Film Tolerances Sheet. After use, remove the oil with alcohol.

No. OptiCell growth chamber is only 2 mm thick that is compatible with regular up-right microscopes as well as inverted ones.

Please check out the protocol of Giemsa/Wright staining in OptiCell under the Applications. When performing an in situ staining in OptiCell, due to the repeat injections through the ports, the ports may be pulled loose and cause leak. Simply push the ports down tightly into the slots will solve this problem.

No. Xylene will damage OptiCell membrane and should not be used with OptiCell.

No. The current OptiCell magnet works well with Dynabeads®-antibodies (protocol). We are developing a new magnet that will be compatible with other manufacturers such as Miltenyi (MACS) bead conjugated antibodies as well.

If the adhesive surface is kept clean and dry, the OptiCell magnet can be applied and removed several times. Replace the film slip-sheet over the magnet adhesive surface when not in use. When the magnet no longer adheres satisfactorily, use a new magnet. While the OptiCell magnet adhesive is safe for use as directed on the OptiCell growth surfaces, other adhesives may cloud or damage the growth surface and should not be used with OptiCell.

The OptiCell magnet adhesive is safe for use as directed on the OptiCell growth surfaces. Other adhesives may cloud or damage the growth surface and should not be used with OptiCell.