CN204918558U - A hollow cover for freezing deposit solid culture filamentous fungi - Google Patents

Abstract

The utility model relates to a hollow cover for freezing deposit solid culture filamentous fungi and freeze the method of depositing belongs to culture presevation technical research field. The utility model discloses an improvement on the method of the depositing basis of freezing at current solid culture filamentous fungi. Improve and a little lie in: adopt the diameter size between 3mm-6mm, the top facing hollow cover that is covered with the quil with wait to keep the filamentous fungi and co -culture at solid culture base surface, it covers the top facing of hollow cover to grow to the filamentous fungi, the hollow cover of waiting to keep the filamentous fungi with the cover moves to be accomplished the bacterial in the liquid and freezes and deposit to freezing to deposit. The utility model discloses a top facing is covered with the hollow cover of quil and makes according to resin material for the conventional method. The utility model has the advantages of effectively avoided the fungus piece to the solid medium carry and prepare the pollution that the fungus suspension probably brought, convenient operation has, low cost, and can satisfy the advantage of special cultivation requirement provides technical support for keeing of filamentous fungi.

Description

A kind of hollow cap for frozen solid culture filamentous fungus

Technical field

The utility model belongs to cell preservation technique research field, is specifically related to a kind of hollow cap for frozen solid culture filamentous fungus and cryopreservation methods.

Background technology

Filamentous fungus is important microbial resources, and the filamentous fungus of many monoids is scientific research or production bacterial strain, creates material impact, have larger social economic value to human being's production life.Not only species diversity is protected to the preservation of filamentous fungus strain resource, more human research and utilize these bacterial classifications to serve vital role.At present, the preservation of microorganism mainly contains following four kinds of methods: (1) nitrogen ultra low temperature preserving process, and this method is applicable to the preservation of liquid culture and solid culture simultaneously; (2) lyophil preservation method, though this method is applicable to the preservation of liquid and solid culture, two class cultures all need first to be prepared into the bacteria suspension meeting preservation requirement; (3)-80 DEG C of sharp freezing preserving processes, the operation of this method is similar with nitrogen ultra low temperature preserving process, is only that storage conditions (mainly temperature) is different; (4) regularly transplant preserving process, this fado is used for the short term storage of solid culture, and needing, regularly to bacterial classification recovery and preservation again, wastes time and energy.Wherein, nitrogen ultra low temperature preserving process and-80 DEG C of sharp freezing preserving processes are because operating the simple and easy and preservation time longer preservation being widely used in each quasi-microorganism.

For filamentous fungus many employings solid culture mode of preservation, main because solid culture is easy to the observation of fungal growth characteristic, be convenient to find to pollute, thus improve the quality of preservation.At present, the preservation carrying out solid culture filamentous fungus is all draw to get or picking bacterium block, or be then prepared into bacteria suspension carries out frozen.The MICROBANK of PRO-LABDIAGNOSTICS company exploitation for being also according to aforesaid operations during solid culture filamentous fungus frozen, need first will to draw the bacterium block got or bacteria suspension adds in frozen solution, and then add porose pearl to mix, cell attachment is in the inside and outside surface of porose pearl.All there is certain defect and limitation in this kind of cryopreservation methods, mainly because: 1. draw get or the bacterium block of picking for culture presevation time, the substratum that bacterium block carries is present in frozen storing liquid, these substratum being brought in new substratum when carrying out bacterial classification recovery, can have a negative impact to there being the spawn culture of particular requirement; 2. the filamentous fungus of solid culture Pollution risk when preparing bacteria suspension is comparatively large, easily causes preservation failure.

Cryopreservation methods at present about filamentous fungus has been reported, but by literature search, does not find to report with the utility model content phase open source literature roughly the same.

Utility model content

The purpose of this utility model is the deficiency improving prior art, a kind of hollow cap for frozen solid culture filamentous fungus is provided and uses the easy to operate, with low cost of this hollow cap and the cryopreservation methods of solid culture filamentous fungus that specific culture requires can be met, for the preservation of filamentous fungus provides technical support.

The utility model first aspect relates to a kind of hollow cap 3 for frozen solid culture filamentous fungus, and bottom it, 34 are a plane and its inside has be communicated with described bottom 34 hollow 33.

In the preferred embodiment of the utility model, described hollow cap 3 is hemispherical hollow cap, round table-like hollow cap, cylindric hollow cap, coniform hollow cap or rectangular-shaped hollow cap.

In the preferred embodiment of the utility model, the cover 32 of described hollow cap 3 is furnished with quil 31.The shape of this quil 31 is unrestricted, vertical, bending or racemosus wooden fork shape.

In the preferred embodiment of the utility model, the length of described quil 31 is 1mm-4mm.

In the preferred embodiment of the utility model, the diameter of described hollow cap 3 is between 3mm ~ 6mm.

In the preferred embodiment of the utility model, described hollow cap 3 is made up of resin material.

The utility model second aspect relates to a kind of cryopreservation methods for solid culture filamentous fungus, and the method comprises the steps:

A. prepare the dull and stereotyped also inoculation of solid medium and treat preservation filamentous fungus;

B. the hollow cap of sterilising treatment according to the utility model first aspect;

C. the solid culture primary surface treated around preservation filamentous fungus vaccination will be placed in downwards bottom described hollow cap;

D. treat preservation filamentous fungus described in cultivating, cover the quil on the cover of described hollow cap or described hollow cap cover to filamentous fungus growth, obtain long have described in treat the hollow cap of preservation filamentous fungus;

E. treat that the hollow cap 3 of preservation filamentous fungus is transferred in frozen solution described in above-mentioned length being had, after mixing, treat that preservation filamentous fungus strain is frozen by described.

Wherein, equal employing the art routine operation such as the dull and stereotyped preparation of the solid medium that relates in this cryopreservation methods, sterilizing, strain inoculation, spawn culture, bacterial classification are frozen, repeats no more herein.

In the preferred embodiment of the utility model, in above-mentioned steps c, described hollow cap and describedly treat that the distance of preservation filamentous fungus vaccination is 0.3cm ~ 1.0cm.

In the preferred embodiment of the utility model, described in the described length shifted has, treat that the quantity of the hollow cap of preservation filamentous fungus is 4-6 in step e.

When frozen filamentous fungus needs to carry out recovery, directly take out described hollow cap after conveniently operating recovery bacterial classification and be seeded to corresponding substratum.

Compared with prior art, the beneficial effects of the utility model are:

The quil 31 that the cover 32 of 1, the described hollow cap 3 for frozen solid culture filamentous fungus of the present utility model is covered with can either play the effect of attachment mycelia, the effect of transfer mycelia can be played again, the liquid that can be directly used in bacterial classification is frozen, do not need to draw and get or picking bacterium block and prepare the operations such as bacteria suspension, when effectively prevent frozen, bacterium block carries and prepares to solid medium the pollution that bacteria suspension may bring.

2, the described hollow cap 3 for frozen solid culture filamentous fungus of the present utility model is made up of resin material, directly can carry out high-temperature sterilization, and can Reusability, and environmental protection is inexpensive.

3, cryopreservation methods of the present utility model have easy to operate, with low cost and can meet specific culture require advantage, for the preservation of filamentous fungus provides technical support.

Accompanying drawing explanation

Fig. 1 is the structural representation of the hollow cap for frozen solid culture filamentous fungus of the present utility model.

Fig. 2 be filamentous fungus on solid medium with the schematic diagram of described hollow cap Dual culture.

Wherein, each Reference numeral has following implication:

1-solid medium; 2-treats preservation filamentous fungus vaccination; 3-hollow cap; 4-is dull and stereotyped; 31-quil; 32-cover; 33-is hollow; Bottom 34-.

Embodiment

Further illustrate content of the present utility model below in conjunction with the drawings and specific embodiments, but should not be construed as restriction of the present utility model.When not deviating from the utility model spirit and essence, the amendment do the utility model method, step or condition or replacement, all belong to scope of the present utility model.

Embodiment 1: flavus frozen

1. prepare Sabouraud glucose agar, pour in Bechtop after autoclave sterilization in appropriate above-mentioned substratum to aseptic flat board, treat its naturally cooling, obtain Sabouraud glucose agar solid plate;

2. inoculate flavus on described Sabouraud glucose agar solid plate from cryopreservation tube or solid medium flat board;

3. with tweezers, the described hollow cap after sterilizing is positioned over around flavus vaccination, obtains Dual culture flat board (as shown in Figure 2); Wherein said hollow cap diameter 4mm, the distance of described hollow cap and described flavus vaccination is 1.0cm; Wherein, as shown in Figure 1, it is hemispherical hollow cap to the hollow cap 3 used, and it is made up of resin material, and bottom it, 34 are a plane and its inside has be communicated with described bottom 34 hollow 33, and its cover 32 is furnished with quil 31.

4. described Dual culture flat board is positioned in 26 DEG C of constant incubators and cultivates 3 days ~ 5 days, be covered to the quil on described hollow cap cover to mycelial growth, obtain the hollow cap being coated with mycelia;

5. drawing 1mL15% (v/v) glycerine frozen storing liquid adds in cryopreservation tube, then gets with aseptic nipper the hollow cap being coated with mycelia described in 4 ~ 6 and puts into frozen storing liquid;

6. cover the pipe lid of described cryopreservation tube, after mixing for several times, described cryopreservation tube is put into program temperature reduction box gently, carry out follow-up frozen operation and complete the frozen of bacterial classification.

Embodiment 2: Paecilomyces lilacinus frozen

1. prepare potato dextrose agar, pour in Bechtop after autoclave sterilization in appropriate above-mentioned substratum to aseptic flat board, treat its naturally cooling, obtain potato dextrose agar solid plate;

2. inoculate Paecilomyces lilacinus on described potato dextrose agar solid plate from cryopreservation tube or solid medium flat board;

3. with tweezers, the described hollow cap after sterilizing is positioned over around Paecilomyces lilacinus vaccination, obtains Dual culture flat board; Wherein said hollow cap diameter 3mm, the distance of described hollow cap and described Paecilomyces lilacinus vaccination is 0.5cm;

4. be positioned over by described Dual culture flat board in 26 DEG C of constant incubators and cultivate 3 days ~ 5 days, the quil be covered on described hollow cap cover to mycelial growth is empty, obtains the hollow cap being coated with mycelia;

5. 1mL15% (v/v) glycerine frozen storing liquid is added in cryopreservation tube, then get with aseptic nipper the hollow cap being coated with mycelia described in 4 ~ 6 and put into frozen storing liquid;

6. cover the pipe lid of described cryopreservation tube, after mixing for several times, described cryopreservation tube is put into program temperature reduction box gently, carry out follow-up frozen operation and complete the frozen of bacterial classification.

Wherein, the flavus (Aspergillusflavus) related in the utility model embodiment and Paecilomyces lilacinus (Paecilomyceslilacinus) are common filamentous fungus, can buy obtain from Spawn preservation organization both domestic and external.

Claims (5)

1. one kind for the hollow cap (3) of frozen solid culture filamentous fungus, it is characterized in that, bottom it, (34) are a plane and its inside has hollow (33) that are communicated with described bottom (34).

2. hollow cap according to claim 1, is characterized in that, it is hemispherical hollow cap, round table-like hollow cap, cylindric hollow cap, coniform hollow cap or rectangular-shaped hollow cap.

3. hollow cap according to claim 1 and 2, is characterized in that, the cover (32) of described hollow cap is furnished with quil (31).

4. hollow cap according to claim 3, is characterized in that, the length of described quil (31) is 1mm-4mm.

5. hollow cap according to claim 1, is characterized in that, the diameter of described hollow cap (3) is between 3mm ~ 6mm.