CN108572101A - liquid-phase molecular sieve - Google Patents

Abstract

The present invention relates to the new methods for extracting cell sample under a kind of normal temperature and pressure efficiently from mucus, belong to medical field, are suitable in medicine or biological field, the high efficiency extraction cell sample from sputum, cervical samples isorheic sample.Present invention application specific protein, can form the three-dimensional structure of similar molecular sieve in aqueous solution, this technology is named as " Liquid-Phase Molecular Sieve " by inventor." Liquid-Phase Molecular Sieve " does not crack the strand of mucus albumen, and mucin strand is disperseed along longitudinal direction, mucin strand itself or complete, category physical process, and then the cell wrapped in mucus is separated.The present invention quickly and efficiently can extract cell sample from a large amount of mucus at normal temperatures and pressures, and for whole process without chemical reaction, the also variation without temperature and pressure, cell sample has obtained complete protection.The present invention has filled up the technological gap that cell sample is extracted from mucus sample.

Description

Liquid-Phase Molecular Sieve

Technical field

The present invention relates to the new methods for extracting cell sample under a kind of normal temperature and pressure efficiently from mucus, belong to medical field, Suitable in medicine or biological field, the high efficiency extraction cell sample from sputum, cervical samples isorheic sample.

Background technology

The sample containing mucus or a large amount of mucus such as sputum, cervical exfoliated cell, is pathocytology or cell culture etc. The sample type that field frequently encounters.The common feature of such sample exactly wraps effective cell sample in mucus, and And mucus itself, without diagnosis or scientific research value, its own is also great disturbing factor.Therefore, mucus removed and by cell Sample high efficiency extraction comes out, and is the premise and basis that such sample carries out diagnosis or action of scientific research.

Mucus, also known as mucus are a kind of wet and slippery liquid secreted by the mucous layer of human body or the goblet cell of submucosa, Generally jelly.Mucus contains a large amount of mucin, also contains inorganic salts and water.

In fact, the processing to mucus sample that the present invention is signified, needs to reach two targets：First, removal mucus is dry It disturbs, second is that extraction wraps cell sample wherein.

Mucus sample is handled, at present international popular, also almost unique method, is " mucin cracking process ".That is, sharp Slowly mucin is decomposed under conditions of being stirred continuously or heating with reagents such as DDT or sodium hydroxides, is grown Strand is cracked into short molecular chain.Since the strand of mucin is very long, intermolecular molecular force is again very big, therefore, cracks Journey gesture necessarily a large amount of chemical energy, namely why current " mucin cracking process " energy consumption is big, the time is long and imitates for this Fruit also poor reason.Importantly, this " mucin cracking process " is cracking due to the use of the biochemical reagents of higher concentration During mucin, cell sample is also largely destroyed, the final purpose of collection of cellular samples is not achieved.

The above fact is not difficult to find out that current technology, decomposition mucus is all undesirable, just let alone extracts cell sample .Therefore, it there is no effective method well so far in the world while accomplishing not only to decompose mucus this side of collectable cell again simultaneously The work in face especially handles the sputum seldom sample of cell containing a large amount of mucus in this way, has become internationally recognized Problem.

Through research of technique for many years, inventor has searched out the completely new approach effectively to solve the above problems finally.

The key of breakthrough is the transformation of thinking.The processing of such sample, core objective are effective extraction cells, and mucus is made It is not the emphasis of concern for valueless composition.If that is, try every possible means to efficiently extract out by cell sample, It need not have to handle mucus albumen molecule.Under the guidance of new approaches, inventor around cracking mucus problem, by research of technique Focus on and how to extract cell sample in terms of.

Inventor is the study found that in mucus, and cell sample is not to be incorporated into one with mucin, and only by mucin Package, cell are independently of except mucin itself, actually separated state.This is similar to, and fishing net is only by fish packet It is rolled in the inside, is not to be combined into composite organisms with fish.It, have to not be by fishing net to fish frees from fishing net Cable is cracked into the broken line of segment, as long as but fishing net scattering and can be taken off fish.

Through numerous studies and practice, " Liquid-Phase Molecular Sieve " technology of inventor's application innovation has well solved efficient place It manages mucus and effectively extracts the problem of cell sample.

Invention content

Screening and lots of comparing experiments of the inventor through multiple technologies scheme, appliable plant albumen and cellulose are raw material, Extraction has synthesized a kind of albumen of long chain, names as " mucus rabphilin Rab ".This albuminoid is white in the dry state It is powdered, it is nontoxic.When soluble in water, hydrophilic pole long chain can be formed with water molecules, and can be stably dispersed in In entire liquid phase.There are three features for this protein molecular chain tool.First, the side shoot of strand specific structure can be specific with aqueous solution Inorganic salts combine, and form reticular structure staggeredly, about 100-300 microns of mesh diameter.Second is that this reticular structure and mucus albumen It is more affine, and with the protein ingredient of cell membrane compared with alienation.Third, aforementioned two kinds of characteristics only just have in the liquid phase of aqueous solution, And the forming process of reticular structure is irreversible.To sum up feature contains the aqueous solution of " mucus rabphilin Rab " and specific inorganic salts, shape At the three-dimensional structure of similar molecular sieve, this technology is named as " Liquid-Phase Molecular Sieve " by inventor.

" Liquid-Phase Molecular Sieve " processing mucus sample process be：

It is protein molecular chain first further combined with hydrone after mucus sample enters in the liquid of " Liquid-Phase Molecular Sieve ", The strand of mucus albumen is in pencil and diametrically expands, and structure becomes more to disperse.It shakes through external force, is such as shaken with hand Or with swirl mixing device mixing, the strand unicom of the mucus protein of getting one's things ready of expansion wraps cell therein and is further dispersed in " Liquid-Phase Molecular Sieve " in.Then, the characteristic due to " Liquid-Phase Molecular Sieve " with " close mucin and dredge cell membrane ", using molecular force, The strand of mucin is pulled open.Note that the strand transverse breakage of mucin is not opened, by mucin strand edge Longitudinal dispersion, mucin strand itself or complete.Due to the strand of mucin be internal combustion obtain it is very secured, and It is physical entanglement between strand, therefore is in filament shape almost needing no outside energy by being drawn around the mucin strand of fasciculation Amount, relying solely on " Liquid-Phase Molecular Sieve " molecular force of itself can fast implement.At the same time, cell is released from the state wrapped It releases, is suspended in the gap of " Liquid-Phase Molecular Sieve " and discrete mucin strand.Finally, by sample carry out low speed from The heart, under the influence of centrifugal force, cell sample are fully deposited on centrifuge tube tube bottom, and " Liquid-Phase Molecular Sieve " and discrete glutinous egg White strand still stops in the liquid phase.So far, the high efficiency extraction of cell sample is completed.

Actual use the result shows that, " Liquid-Phase Molecular Sieve " technology of the invention can at normal temperatures and pressures, quickly and efficiently Cell sample is extracted from a large amount of mucus, whole process is without chemical reaction, and also the variation without temperature and pressure, cell sample obtain Complete protection.

Specific implementation mode

The present invention has lower key point in implementation process：

1) this technology is very sensitive to low temperature and high temperature.

Suitable temperature limit is -20 DEG C to 60 DEG C.But, under common use environment, such as clinical unit It is what there is no problem between routine operation etc..It should avoid freezing in storage and transport process.

2) centrifugal force can not be excessive, and otherwise the separating effect of cell sample can be affected.

Commercially available low speed medical centrifugal machine is ideal chose, and rotating speed is generally at 4000 revs/min or less.Unusable high speed Centrifuge or refrigerated centrifuge.The setting of other kinds of centrifugation apparatus, centrifugal force should refer to low speed medical centrifugal machine.

3) prepared " Liquid-Phase Molecular Sieve " liquid, can be miscible with the reagents such as plurality of inorganic salt or alcohols.

4) ultraviolet source of prepared " Liquid-Phase Molecular Sieve " liquid, daylight or disinfection does not interfere with its performance, but answers Avoid long-time direct projection.

5) at suitable temperature and condition of storage, prepared " Liquid-Phase Molecular Sieve " liquid, after sealing, the term of validity is at least It is 26 months.Whether the longer time is effective, ends the applying date, and inventor not yet further studies.

Using effect

The present invention has filled up the technological gap that cell sample is extracted from mucus sample.

After actual use shows clear this entrance of sputum sample " Liquid-Phase Molecular Sieve " liquid, only need 5 minutes or so, you can will wherein 90% or more cell centrifugation extracts.Even dense sputum sample sheet, into after " Liquid-Phase Molecular Sieve " liquid, most 25-30 points Clock, you can extract wherein 90% or more cell centrifugation.

Description of the drawings

Fig. 1 is the molecular structure model figure of " Liquid-Phase Molecular Sieve " affine discrete mucin strand.Layer distributed simultaneously has rule Then network is " Liquid-Phase Molecular Sieve " structural representation, and it is mucin molecular chain structure to intert irregular micel therein Schematic diagram.

Fig. 2 is using this technology treated dense sputum sample sheet, and the cell mass after centrifugation is extracted has carried out cytologic slide, 20 The design sketch of times biologic photomicroscope.The result shows that, the cell of extraction is handled through this technology, form is complete, and structure is clear under mirror It is clear, it has obtained protecting to greatest extent.Moreover, smear clean background, no mucus residual, thoroughly eliminate the dry of mucus very much It disturbs.

Claims (4)

1. A kind of 1. " Liquid-Phase Molecular Sieve ", which is characterized in that the three-dimensional structure of similar molecular sieve can be formed in aqueous solution.
2. A kind of 2. " Liquid-Phase Molecular Sieve ", which is characterized in that the strand for not cracking mucus albumen, by mucin strand Disperse along longitudinal direction, mucin strand itself or complete.
3. A kind of 3. " Liquid-Phase Molecular Sieve ", which is characterized in that during the entire process of processing mucus, no chemical reaction, also without temperature and The variation of pressure.
4. A kind of 4. " Liquid-Phase Molecular Sieve ", which is characterized in that during the entire process of processing mucus, cell sample is not destroyed itself, Cellular prion protein is complete.