CN101485303A - Exfoliated cell preservative fluid - Google Patents

Abstract

The invention relates to a preservation solution for exfoliated cells, which is characterized by comprising a pH buffering agent, an osmotic pressure maintaining agent, an anti-corrosion antistaling agent, a fixative for maintaining cellular morphologies, an anticoagulant, a mucus softener, proteinase K and a component for destroying red blood cells. Compared with the prior art, the preservation solution has the advantages that the cell storage rate is high, the storage time at normal temperature is long, the mucus is sufficiently dissolved and the red blood cells are partially destroyed, the sheet-making effect comprises quite even cell distribution, the cellular morphologies are good, the cytolymph and the cell nucleus are divided clearly and are well arranged, the brightness of the cytolymph and the cell nucleus is quite good, and the karyoplasms, the morphological structures and the amount of the cells are quite clear.

Description

A kind of exfoliated cell preservative fluid

Technical field

The present invention relates to a kind of exfoliated cell preservative fluid.

Background technology

Living cells has physiological function, biologic activity and intrinsic cellular morphology and structure, has using value aspect a lot.But, it places if being broken away from original living environment, cells physiological function, biologic activity and intrinsic cellular morphology and structure will change, conventional method is exactly that the cell that comes off is carried out freezing preservation, and freezing preservation needs special freezing equipment, relatively more expensive, and, owing to there is the harm of pair cell damage in freezing, the course of defrosting, thereby cause cytometaplasia or distortion, influence follow-up work.

Thus, the solution that cast-off cells are preserved has appearred being specifically designed to, as the preservation liquid (PresetverCyt of the new Bai Shi of the U.S. (Tinpreper TCT) production on the market ^TM), this preservation liquid is and the matching used reagent of machine Tinpreper-2000, costs an arm and a leg.For reducing application cost, the researcher has developed a kind of new preservation formula of liquid, application reference number is open " exfoliated cell preservative fluid " (publication number: CN1834233A) of Chinese patent application of 200610017591.9, the preservation liquid of this application is grouped into by following one-tenth, by volume part meter: 5 parts～10 parts of 40 parts～60 parts of methyl alcohol or ethanol, phosphate buffer or 20 parts～40 parts of trishydroxymethylaminomethane hydrochloride buffers, formaldehyde or 5 parts～10 parts in acetone, EDTAP dipotassium ethylene diamine tetraacetate or disodium ethylene diamine tetraacetates.

The patent documentation of this technical field again can application reference number be open " aqueous solution for cell preservation " (publication number: CN101310009A) of Chinese patent application of 200680042980.5, this aqueous solution comprises thickener, antifreezing agent, sugar, but do not contain natural animal source component, as serum and basal medium etc.Can also application reference number be that (publication number: CN101182506A), exfoliated cell preservative fluid comprises the component of following proportioning in parts by volume: methyl alcohol 10-15 part for 200710050463.9 Chinese patent application open " cast-off cells are preserved the preparation method of fixer and cell thin smear "; Ethanol 10-15 part; Glycerine 1-2 part; Polyethylene glycol 1-2 part; Formaldehyde 10-12 part; Glacial acetic acid 0.5-1 part; Mass concentration is sodium chloride 10-20 part of 0.85-0.9%; Phosphate buffer 33-68.5 part.This preservation liquid has intrinsic effective, the characteristics that cell bonding die rate is high of cell.

Summary of the invention

Technical problem to be solved by this invention is that a kind of cell storage rate height, long exfoliated cell preservative fluid of holding time are provided at the above-mentioned state of the art.

The present invention solves the problems of the technologies described above the technical scheme that is adopted: a kind of exfoliated cell preservative fluid is characterized in that comprising that pH buffer, osmotic pressure keeps agent, antisepsis antistaling agent, is used to keep fixative, anticoagulant, mucus softening agent, the humectant of cellular morphology and destroys erythrocytic composition.

As preferably, described pH buffer pH value is 6.5～7.4.

As preferably, at least a among the PBS that described pH buffer is, Tris HCl or the PIPES;

It is sodium chloride solution that described osmotic pressure is kept agent;

Described antisepsis antistaling agent is at least a in benzoic acid sodium salt, the benzoic acid sylvite;

Described fixative is at least a in absolute ethyl alcohol, methyl alcohol, paraformaldehyde, acetone, the coumarin;

Described anticoagulant is that ethylenediamine tetra-acetic acid two is received, at least a in the EDTAP dipotassium ethylene diamine tetraacetate, heparin;

Described mucus softening agent is a Proteinase K;

Described humectant is a carbolic acid;

The erythrocytic composition of described destruction is at least a in sodium bicarbonate, the tertiary sodium phosphate.

As preferably, comprise following component and percentage by weight:

Sodium chloride 0.5%～0.9%

Sodium bicarbonate 0.1%～0.5%

Tertiary sodium phosphate 0.1%～0.5%

Anticoagulant 5%～10%

Antisepsis antistaling agent 1%

Proteinase K: 0.1～0.5%

Paraformaldehyde 1%～4%

Coumarin 1%～4%

Carbolic acid 1%～5%,

Also be added with following component and percent by volume ratio:

PH buffer 50%～70%

Absolute ethyl alcohol 12%～20%

Methyl alcohol 10%～15%

Acetone 1%～4%,

Wherein, described anticoagulant is that ethylenediamine tetra-acetic acid two is received, a kind of in the EDTAP dipotassium ethylene diamine tetraacetate, heparin, described antisepsis antistaling agent is a kind of in benzoic acid sodium salt, the benzoic acid sylvite, and described pH buffer is a kind of among PBS, Tris HCl or the PIPES.

Compared with prior art, the invention has the advantages that: preserve each component rational proportion in the liquid, collaborative mutually, make cell storage rate height, can preserve length under the normal temperature, the longlyest can reach 2 years; Can fully dissolve mucus and partial destruction red blood cell; It is very even that production effect has cell distribution, cellular morphology is intact, and endochylema and cell nucleus boundary are clear, well arranged, and the bright property of endochylema and cell nucleus is very good, the morphosis and the quantity of the very clear cell of caryoplasm can be used for HPV-DNA, Chlamydia and SABC test simultaneously; Can be provided for the special-purpose liquid base of other position cytolgical examinations; Same sample can repeat film-making repeatedly, and has homogeneity; Cell-preservation liquid is nontoxic pollution-free, guarantees operator ' s health safety; Fully kept the cell component that diagnostic significance is arranged; Deployment cost is lower, is easy to promote.

Embodiment

Embodiment describes in further detail the present invention below in conjunction with accompanying drawing.

Embodiment 1: the exfoliated cell preservative fluid in the present embodiment comprises as follows:

Components in weight percentage: 0.7% sodium chloride, 0.25% tertiary sodium phosphate, 7.5% ethylenediamine tetra-acetic acid two is received, one of in the EDTAP dipotassium ethylene diamine tetraacetate, heparin, 1% benzoic acid is received salt (or sylvite), 3% paraformaldehyde, 3% coumarin, 0.1% Proteinase K, 0.3% sodium bicarbonate, 2.5% carbolic acid.

Components in percentage by volume: 60%pH buffer (pH7), 16% absolute ethyl alcohol, 12.5% methyl alcohol, 3% acetone, wherein, the pH buffer is a kind of among PBS, Tris HCl, the PIPES.

Embodiment 2: the exfoliated cell preservative fluid in the present embodiment comprises as follows:

Components in weight percentage: 0.9% sodium chloride, 0.1% tertiary sodium phosphate, 10% ethylenediamine tetra-acetic acid two is received, one of in the EDTAP dipotassium ethylene diamine tetraacetate, heparin, 1% benzoic acid is received salt (or sylvite), 4% paraformaldehyde, 4% coumarin, 0.5% Proteinase K, 0.5% sodium bicarbonate, 1% carbolic acid.

Components in percentage by volume: 50%pH buffer (pH6.5), 20% absolute ethyl alcohol, 15% methyl alcohol, 4% acetone, wherein, the pH buffer is a kind of among PBS, Tris HCl, the PIPES.

Embodiment 3: the exfoliated cell preservative fluid in the present embodiment comprises as follows:

Components in weight percentage: 0.5% sodium chloride, 0.5% tertiary sodium phosphate, 5% ethylenediamine tetra-acetic acid two is received, one of in the EDTAP dipotassium ethylene diamine tetraacetate, heparin, 1% benzoic acid is received salt (or sylvite), 1% paraformaldehyde, 4% coumarin, 0.3% Proteinase K, 0.1% sodium bicarbonate, 5% carbolic acid.

Components in percentage by volume: 70%pH buffer (pH7.4), 12% absolute ethyl alcohol, 10% methyl alcohol, 2% acetone, wherein, the pH buffer is a kind of among PBS, Tris HCl, the PIPES.

Result of the test shows, the exfoliated cell preservative fluid cell storage rate height of embodiment 1, embodiment 2 and embodiment 3, holding time are long, can preserve under the normal temperature 2 years.

Claims (4)

1, a kind of exfoliated cell preservative fluid is characterized in that comprising that pH buffer, osmotic pressure keeps agent, antisepsis antistaling agent, is used to keep fixative, anticoagulant, mucus softening agent, the humectant of cellular morphology and destroys erythrocytic composition.

2, exfoliated cell preservative fluid according to claim 1 is characterized in that described pH buffer pH value is 6.5～7.4.

3, exfoliated cell preservative fluid according to claim 2 is characterized in that

At least a among the PBS that described pH buffer is, Tris HCl or the PIPES;

It is sodium chloride solution that described osmotic pressure is kept agent;

Described antisepsis antistaling agent is at least a in benzoic acid sodium salt, the benzoic acid sylvite;

Described fixative is at least a in absolute ethyl alcohol, methyl alcohol, paraformaldehyde, acetone, the coumarin;

Described anticoagulant is that ethylenediamine tetra-acetic acid two is received, at least a in the EDTAP dipotassium ethylene diamine tetraacetate, heparin;

Described mucus softening agent is a Proteinase K;

Described humectant is a carbolic acid;

The erythrocytic composition of described destruction is at least a in sodium bicarbonate, the tertiary sodium phosphate.

4, exfoliated cell preservative fluid according to claim 3 is characterized in that comprising following component and percentage by weight:

Sodium chloride 0.5%～0.9%

Sodium bicarbonate 0.1%～0.5%

Tertiary sodium phosphate 0.1%～0.5%

Anticoagulant 5%～10%

Antisepsis antistaling agent 1%

Proteinase K: 0.1～0.5%

Paraformaldehyde 1%～4%

Coumarin 1%～4%

Carbolic acid 1%～5%,

Also be added with following component and percent by volume ratio:

PH buffer 50%～70%

Absolute ethyl alcohol 12%～20%

Methyl alcohol 10%～15%

Acetone 1%～4%,

Wherein, described anticoagulant is that ethylenediamine tetra-acetic acid two is received, a kind of in the EDTAP dipotassium ethylene diamine tetraacetate, heparin, described antisepsis antistaling agent is a kind of in benzoic acid sodium salt, the benzoic acid sylvite, and described pH buffer is a kind of among PBS, Tris HCl or the PIPES.