CN201867329U

CN201867329U - Reaction tube for preparing cell block

Info

Publication number: CN201867329U
Application number: CN2010205026503U
Authority: CN
Inventors: 杨琨; 宋朝君; 金伯泉; 魏玉英; 孙元杰
Original assignee: Fourth Military Medical University FMMU
Current assignee: Fourth Military Medical University FMMU
Priority date: 2010-08-24
Filing date: 2010-08-24
Publication date: 2011-06-15
Anticipated expiration: 2020-08-24

Abstract

The utility model discloses a reaction tube for preparing a cell block, which comprises a thin-walled pointed plastic reaction tube, and an agar gel block is laid at the bottom of the plastic reaction tube. By laying the agar gel block at the bottom, cells can be separated and precipitated on the gel block when a cell mass is prepared, and after the cell mass is formed, the bottom of the reaction tube is excised, and the gel block and the cell mass are ejected from an opening together, thereby enabling the shape of the formed cell mass to be complete; and the cell mass with complete shape can be obtained as long as the cell mass with obvious lamination and the gel block are peeled off, thereby overcoming the defect that the separation of the cell mass is very easy to break usually, and enabling the success rate to be as high as 100%.

Description

A kind of reaction tube that is used to prepare the cell wax stone

Technical field

The utility model belongs to the cell engineering field, relates to the preparation of cell wax stone, particularly a kind of reaction tube that is used to prepare the cell wax stone.

Background technology

Along with finishing of " Human Genome Project ", the mankind enter " functional genome's plan ", and proteomics is the important research content of " functional genome's plan ".Protein structure and function diversity have determined the complicacy of proteomics research, the protein expression regularity of distribution of especially new coded by said gene, the announcement of 26S Proteasome Structure and Function to the biologist be rich in challenging.The most frequently used method of the research protein expression regularity of distribution is the immunohistochemical staining technology, it be utilize specific antibody in the histocyte original position by antigen-antibody reaction and histochemical color reaction, corresponding antigens is carried out a technology of qualitative, location, quantitative measurement.The cell wax stone is being played the part of more and more important role in immunohistochemical staining.

The cell wax stone is the cell centrifugation in the cell of in vitro culture, primary cell or the humoral specimens such as hydrothorax, ascites to be collected form cell mass, make after conventional dehydration after fixing, waxdip, the embedding, but its advantage is to can be made into many sections and long preservation, carry out a series of histochemistries and immunohistochemical staining, or carry out experiments such as Electronic Speculum, in situ hybridization, all have great importance at the aspects such as diagnosis and differential diagnosis of location, expression and the disease of cell and subcellsular level for the research specified protein.

At present the preparation method of cell wax stone commonly used is broadly divided into two kinds of centrifuge method and agar methods.It is earlier cell centrifugation to be collected the centrifuge tube bottom that centrifuge method prepares the cell wax stone, manages cell mass is taken out after fixing again, the conventional afterwards cell wax stone of making.The shortcoming of this method is that the cell quantity that needs is bigger, is difficult for the cell quantity that reaches enough for primary cell or clinical samples, moreover, take out cell mass and cause agglomerate to break easily, then all that has been achieved is spoiled.It is that the agar solution of equivalent and cell suspension are mixed that agar method prepares the cell wax stone, fix after solidifying and the wax stone preparation, its shortcoming is an apart from each other between the cell, like this, in follow-up immunocytochemical stain, edge effect occurs easily and cause false positive results, in addition, cell density is less also judges to the result and takes the photograph sheet and make troubles.Therefore, the preparation method of the cell wax stone of using always at present can't satisfy the requirement of growing scientific research and clinical pathology diagnosis.

The utility model content

The technical matters that the utility model solves is to provide a kind of reaction tube that is used to prepare the cell wax stone, use this reaction tube to prepare the cell wax stone, Shi Buzai needs a large amount of cells, cell mass can be not broken, fit tightly between the cell in the cell wax stone, the power that is prepared into of cell wax stone is greatly improved.

The utility model is to be achieved through the following technical solutions:

A kind of reaction tube that is used to prepare the cell wax stone comprises the plastics reaction tube of thin-walled tip being equipped with the agar gel piece in the bottom of plastics reaction tube.

Described agar gel piece is laid from the bottom to half height of plastics reaction tube.

Described plastics reaction tube is the PCR pipe.

Compared with prior art, the utlity model has following beneficial technical effects:

The design of the utility model by the plastics reaction tube of, thin-walled tip smooth at tube wall be for the ease of the carefulness separation of cell and cell mass with the separating of reaction tube; Lay the agar gel piece that contains sucrose in the bottom, when being used to prepare cell mass, make cell separation be deposited on the gel piece, after cell mass forms, the bottom of excision reaction tube, gel piece and cell mass are ejected from opening part in the lump, make that so formed cell mass shape is complete, as long as will have the cell mass of obvious layering and gel piece peels off, just can access the complete cell mass of shape, overcome common cell mass and separated the defective that is easy to fragmentation, success ratio can reach 100%.

Shape is complete when forming cell mass, and is convenient to separate the just preparation of realization cell mass easily of so only needs control isolated cells concentration, no longer needs a large amount of cells, the preparation of the cell wax stone that especially convenient preparation source is rare.Because the cell mass by centrifugal formation deposition fits tightly between the cell, can get rid of the false positive results that issuable edge effect caused in the dyeing, can satisfy the requirement of growing scientific research and clinical pathology diagnosis.

Description of drawings

Fig. 1 is cell deposition forms cell mass on the agar gel piece a synoptic diagram;

Fig. 2 is the synoptic diagram of the bottom of excision plastics reaction tube with the taking-up cell mass.

Wherein: 1 is the plastics reaction tube; 2 is the agar gel piece; 3 is cell mass.

Embodiment

Below in conjunction with the concrete operations of accompanying drawing and preparation cell wax stone the utility model is done to describe in further detail, the explanation of the invention is not limited.

As shown in Figure 1, a kind of reaction tube that is used to prepare the cell wax stone comprises the plastics reaction tube 1 of thin-walled tip, lays the agar gel piece 2 that contains sucrose in the bottom of plastics reaction tube 1, and the design of thin-walled tip is to separate for the ease of cell centrifugation.Specifically can adopt the PCR pipe to use, according to the thin-walled PCR pipe of how much getting the specification that varies in size of preparation cell quantity, cell quantity more after a little while, the PCR pipe that the capacity of taking is little is with preparation shape cell mass preferably.

Being laid in of the described agar gel piece that contains sucrose: at first get the 1g agar powder, add the 100ml mass concentration and be in 25～30% the sucrose solution, be heated to boiling, it is standby to put 80 ℃ of baking boxs; Then the sucrose of ready heat-agar solution is added in the PCR pipe to half-full, cool off then and in the PCR pipe, form agar gel piece 2, also the i.e. height of institute's agar gel piece 2 that forms from the bottom laying of PCR pipe to half.

After formation agar gel piece 2 is laid in the bottom of PCR pipe, just can carry out the preparation of cell wax stone:

1) cell centrifugation in cell, primary cell or the humoral specimens such as hydrothorax, ascites of in vitro culture is collected, abandoned behind the supernatant resuspendedly, prepare denseer cell suspension (1～2 * 10 with the isotonic liquid of classes such as small amount of H anks liquid, RPMI 1640 nutrient culture media or PBS ⁷/ m1);

2) cell suspension is added in the PCR pipe that forms agar gel piece 2, with the centrifugal 5min of swing bucket rotor 2000r/min, abandon supernatant, cell deposition forms cell mass 3 on agar gel piece 2;

3) above-mentioned PCR pipe is immersed in the neutral formalin solution fixedly 12h;

4) fixedly finish after, take out the PCR pipe; Carefully cut PCR pipe bottom with single-edge blade then and divide (as shown in Figure 2), partly eject from the PCR tube opening together with cell mass 3 with glass rod or the first agar gel piece 2 of sample injector gently, carefully cell mass 3 and agar gel piece 2 agar are separated the cell mass 3 that obtains having complete shape more at last;

5) cell mass that shape is complete 3 is made into cell wax stone and section after according to conventional dehydration, waxdip, embedding.

Claims

1. a reaction tube that is used to prepare the cell wax stone is characterized in that, comprises the plastics reaction tube (1) of thin-walled tip, is equipped with agar gel piece (2) in the bottom of plastics reaction tube (1).

2. the reaction tube that is used to prepare the cell wax stone as claimed in claim 1 is characterized in that, described agar gel piece (2) is laid from the bottom to half height of plastics reaction tube (1).

3. the reaction tube that is used to prepare the cell wax stone as claimed in claim 1 is characterized in that, described plastics reaction tube (1) is the PCR pipe.