CRYOBEADS™

Cat. no. CB12 CryoBeads™, 8ml vial, 20 beads 12 vials/box

INTENDED USE

CryoBeads™ are porous beads, which are intended as carriers to support the viability of microorganisms during storage in the freezer.

This product is not intended to be used for the diagnosis of human disease.

SUMMARY

The need for storing microorganisms for extended periods of time is a common practice in the field of microbiology. Known organisms are used in the laboratory for use in quality control and education. CryoBeads™ offer a superior system for these purposes.

FORMULA

Ingredients per 850ml of deionized water:*

Brucella Broth:
Peptone 20 15.0gm
Peptone 100 5.0gm
Sodium Chloride 5.0gm
Yeast Extract 2.0gm
Dextrose 1.0gm
Sodium Citrate 1.0gm
Sodium Bisulfite 0.1gm
Glycerol 150.0ml

Final pH 7.0 +/- 0.3 at 25ºC.

Each vial contains at least 20 polypropylene beads.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-30ºC. away from direct light. Media should not be used if there are any signs of deterioration (shrinking or discoloration), contamination, or if the expiration date has passed. Protect from excessive heat and freezing.

PRECAUTIONS

PRINCIPLE

Each vial contains approximately 20-30 individual CryoBeads™, along with a cryopreservative liquid (Brucella Broth with Glycerol). Microorganisms readily adhere to the surface because of the beads porous nature. For extended storage, after inoculation, vials are kept frozen. When requiring fresh cultures, a single bead can be removed and utilized to directly inoculate a suitable bacteriological medium.

PROCEDURE

Specimen Collection: Infectious material should be submitted directly to the laboratory without delay and protected from excessive heat and cold. If there is to be a delay in processing, the specimen should be inoculated onto an appropriate transport media and refrigerated until inoculation.

Method of Use:

1. For each vial to be inoculated and stored, obtain a pure isolated culture on plated media. Cultures should be from 18 to 24 hours old.

2. Open the screw cap vial using aseptic technique.

3. Using a sterile loop, transfer growth from the pure culture and inoculate it into the cryopreservative fluid. Twirl loop to disperse the organisms.

4. Mix the microorganisms by closing the vial and inverting a few times. Shake or vortex well. To ensure that the cryoprotectant has penetrated the bacterial cells, allow the suspension to sit at room temperature for a minimum of 15 minutes, but no longer than 45-60 minutes. (6)

5. At this point, the microorganisms will adhere to the porous beads. The fluid is then removed by aspirating the fluid into a sterile pipette using aseptic technique. We suggest our disposable sterile pipet (Cat. no. 22220S). Discard the fluid into a disinfectant solution. Close the vial tightly.

6. Identify the vial with a permanent marking pen.

7. For best long-term results, store inoculated vial at -70ºC. Storage at -20ºC. is acceptable, but organisms will not remain viable for as long a period of time.

Reconstitution Procedure:

1. Remove vial from low temperature storage and move to an aseptic environment when the microorganisms are required for laboratory use.

2. Under aseptic conditions, open the vial and remove one of the colored beads using a sterile nichrome needle (Cat. no. 3060), a disposable plastic needle (Cat. no. HSND), or forceps. Quickly re-cap the vial and return it immediately to the freezer. This task must be performed quickly. Do not allow the contents of the vial to thaw.

3. The inoculated bead is then rolled on an appropriate non-selective, solid, plated medium, such as Chocolate Agar or Blood Agar. A sterile loop is then streaked across the area of inoculation several times to achieve isolated colonies.

Inoculation of the bead into a broth medium is not recommended. Media containing any antibiotics or other inhibitory ingredients is also not recommended. When culturing Campylobacter, it is recommended to use the jar method rather than a bag or pouch.

4. Immediately return the vial to cold storage before it thaws. Thawing and refreezing of the beads will adversely affect the viability of the organisms.

5. Incubate the plates immediately in an appropriate atmosphere (i.e. aerobic, anaerobic, CO 2 , or microaerophilic) and temperature, depending on the organism to be cultured. Allow one to four days for growth to appear.

LIMITATIONS

This product is offered only as a means of providing extended storage possibilities for microorganisms.

CryoBeads™ should not be utilized if any of the following circumstances are present:

a. Prior to use, there is any trace of leakage (loss of cryopreservative fluid) or cracking of the vial.
b. There is turbidity in cryopreservative indicating possible contamination.
c. The date of expiration on the outer label has been exceeded. Expiration date is applicable only for the unopened vial.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, other culture media, incinerators, freezers, incubators, forceps, and sterile pipets, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Escherichia coli
ATCC® 25922
E 24hr 35°C Aerobic Growth on Blood Agar after freezing CryoBeads™ for 48 hours
Streptococcus pyogenes
ATCC® 19615
E 24hr 35°C Aerobic Growth, with beta-hemolysis, on Blood Agar after freezing CryoBeads™ for 48 hours

User Quality Control

PHYSICAL APPEARANCE

CryoBeads™ broth should appear light amber in color, and each vial should contain at least 20 colorless polypropylene beads.

CryoBeads™

CryoBeads™ (Cat. no. CB12).

CryoBeads™

CryoBeads™ (Cat. no. CB12).

REFERENCES

1. Medical Laboratory Sciences. 1985; 42:289-290 (U.K.).

2. Feltham., et al. 1978. Journal of Applied Bacteriology; 44:313-316.

3. Nagel, Janice G., and Cunz, Lawrence J. 1971. Simplified Storage and Retrieval of Stock Cultures. Applied Microbiology; April, Vol. 23, No. 4, p. 837-838.

4. Clinical and Pathogenic Microbiology, Howard ; 1987. p. 54, C.V. Mosby Publishing.

5. " Quality Assurance for Commercial Culture Media Manufacturer ", M22-A2. Clinical Laboratory Standards Institute (CLSI - formerly NCCLS), Villanova, PA.

6. ATCC connection, Vol. 26, No. 1. 2006. p. 4-5.


ATCC is a registered trademark of the American Type Culture Collection.

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