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Glaucoma chattoni Corliss

50034

Product category
Protists
Classification
KINGDOM: Protozoa
Strain designation
GT-1
Type strain
No
Geographical isolation
United States; Tennessee
Product format
Test tube
Storage conditions
See handling procedure
Mission Collection Item
This is a Mission Collection Item

Documentation

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Biosafety Icon BSL 1

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information

Handling information

Medium
Instruction for complete medium
ATCC Medium 802 may be pre-inoculated with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831™) or Enterobacter aerogenes (ATCC® 13048™) for better growth.
Temperature
20-25°C
Atmosphere
Aerobic
Handling procedure
Handling of Live Culture
This strain is routinely shipped as a growing culture in a glass 16 x 125 mm screw-capped test tube.  The volume of the cell suspension is approximately 5 mL.  When the culture arrives remove it promptly from the shipping container.  Do not store the culture at refrigeration temperatures before handling.  To assure viability, immediately loosen the test tube cap and incubate upright at 20-25°C for at least one hour before observing the culture.  There should be numerous active trophozoites in suspension.  If the numbers are low the culture may have been exposed to temperature extremes in transit.  Regardless of the state of the culture, aseptically transfer a 0.5 mL aliquot to a T-25 flask containing 10 mL of ATCC medium 802 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831™) or Enterobacter aerogenes (ATCC® 13048™).  Incubate with the cap tightly sealed at 20-25°C.
Culture maintenance
Subculture at peak density (approximately every 14-21 d) to a fresh T-25 flask of fresh medium in the following manner:
  1. Gently agitate the flask and aseptically transfer 0.25-0.5 mL to a T-25 tissue culture flask containing 10 mL complete medium.
  2. Incubate with the cap tightly sealed at 20-25°C.
Reagents for cryopreservation
Cryoprotective Solution
DMSO, 2.0 mL
Fresh growth medium, 8.0 mL
Cryopreservation


  1. To achieve the best results, set up cultures with several different inocula (i.e., 0.25 mL, 0.5 mL, 1.0 mL).  Harvest cultures and pool when the culture that received the lowest inoculum is at or near peak density.
  2. If the cell concentration exceeds the required level do not centrifuge, but adjust the concentration to approximately 2 x 106 cells/mL with fresh growth medium.  If the concentration is too low, centrifuge at 400-500 x g for 5 min and resuspend the pellet in the volume of fresh medium required to yield the desired concentration.
  3. While cells are centrifuging prepare a 20% (v/v) solution of sterile DMSO as follows:  Add the required volume of DMSO to a glass screw-capped test tube and place it in an ice bath.  Allow the DMSO to solidify.  Add the required volume of refrigerated medium.  Dissolve the DMSO by inverting the tube several times. 
    Note:
    If the DMSO solution is not prepared on ice, an exothermic reaction will occur that may precipitate certain components of the medium.
  4. Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be approximately 106 cells/mL and 10.0% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution to the start of the freezing process should be no less than 15 min and no longer than 30 min.
  5. Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation).
  6. Place the vials in a controlled rate freezing unit.  From room temperature cool at -1°C/min to -40°C.  If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through the heat of fusion.  At -40°C plunge into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus.  Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.)  
  7. The frozen preparations are stored in either the vapor or liquid phase of a nitrogen freezer.
  8. To establish a culture from the frozen state, quickly add 0.5 mL spent/exhausted ATCC medium 802* supplemented with 12% (w/v) sucrose to the frozen ampule and place it in a 35°C water bath.  Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.
    *Spent/exhausted medium is medium which has been previously used for cultivation of food-source bacteria and subsequently filter-sterilized for general use. The bacteria consume many of the nutrients in the medium, reducing its capacity to promote additional bacterial growth when re-used. It is prepared by passing supernatant from an advanced culture (i.e., one which is at or near stationary phase) through a 0.22 μm filter.
  9. Immediately after thawing, do not leave in water bath, aseptically remove the contents of the ampule and inoculate onto the surface of a 20 x 100 mm petri plate containing ATCC medium 919 (non-nutrient agar) with an overlay of 15.0 mL spent ATCC medium 802 supplemented with 6% (w/v) sucrose.
  10. Incubate at 20-25°C with the cap on loosely.
  11. Once the culture is established (motile cells observed), aseptically transfer approximately 8 mL to an upright 20x150mm glass test tube and gently overlay with an equal volume of ATCC medium 802 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831™) or Enterobacter aerogenes (ATCC® 13048™).  The bacterized medium should form a bilayer with the spent medium below.
  12. When cells migrate from the layer of spent medium upward into the layer of bacterized medium, aseptically remove a 0.5-1.0 mL aliquot and transfer to a T-25 tissue culture flask containing 10.0 mL of ATCC medium 802 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831™) or Enterobacter aerogenes (ATCC® 13048™).
  13. Incubate with the cap tightly sealed at 20-25°C.
  14. Follow the protocol for maintenance of culture.

History

Deposited as
Glaucoma chattoni Corliss
Depositors
EM Simon, DL Nanney
Chain of custody
ATCC <-- EM Simon, DL Nanney <-- J.W. McCoy <-- . . .

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

McCoy JW. Updating the tetrahymenids IV. Cortical properties of Glaucoma. Protistologica 40: 149-158, 1975.

McCoy JW. New features of the Tetrahymenid cortex revealed by protargal staining. Acta Protozool. 13: 135-159, 1974.

Nanney DL, et al. Comparison of sequence differences in a variable 23S rRNA domain among sets of cryptic species of ciliated protozoa. J. Eukaryot. Microbiol. 45: 91-100, 1998. PubMed: 9495037

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