CN110411813A - A kind of method cell block sizing fixed plate and make cell block with it - Google Patents
A kind of method cell block sizing fixed plate and make cell block with it Download PDFInfo
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- CN110411813A CN110411813A CN201910533208.2A CN201910533208A CN110411813A CN 110411813 A CN110411813 A CN 110411813A CN 201910533208 A CN201910533208 A CN 201910533208A CN 110411813 A CN110411813 A CN 110411813A
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/36—Embedding or analogous mounting of samples
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2873—Cutting or cleaving
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of methods cell block sizing fixed plate and make cell block with it, belong to cell pathology experimental method technical field, sizing fixed plate provided by the invention may be implemented to fix the good plastotype of cell precipitate, and the utensil structure is simple, easy to use, experimental result confirms, the cell block prepared using method provided by the invention and utensil, cell precipitate condensation is close, and spallation, broken phenomenon are less prone in subsequent operation.Cell block production method provided by the invention is simple, and cell precipitate is embedded in paraffin mass in the form of substantially regular.The present invention has well solved in conventional cell wax stone preparation method that cell precipitate condensation effect is bad, is easy the fragmentation in subsequent operation, leads to the shortcomings that cell block production failure.
Description
Technical field
The invention belongs to cell pathology experimental method technical field, it is related to a kind of cell block manufacturing appliance and production
A kind of method, and in particular to method cell block sizing fixed plate and make cell block with it.
Background technique
Cell pathology is that the important branch of the clinicopathologia of medical diagnosis on disease is carried out by cell in detection body fluid sample
Subject plays an important role in the diagnosis and differential diagnosis of medical diagnosis on disease, especially innocent and malignant tumour.It is generally known that In
In malignant tumour progress and invasion transfer, usually adjoint different degrees of dropsy of serous cavity (such as hydrothorax, ascites) can occur,
And malignant cell and tumour-specific mark are detected from above-mentioned sample using cell pathology technology becomes progress tumour and examines
Disconnected and antidiastole most important, most efficient method.Meanwhile urine, peripheral blood and tumor fine needle puncture the puncture obtained
Object or cyst fluid are also the usual cell pathology detection sample for carrying out diagnosing tumor and antidiastole.But due to body fluid sample
Middle cell quantity is limited, using direct smear, centrifuged deposit object smear or is directly centrifuged traditional flaking method such as smear usually
Sufficient amount of aim cell can not be obtained, it is difficult to meet minimum requirements of the cytodiagnosis to target cell numbers.And
And since sample size is limited, existing method can only also prepare several smears, it is difficult to carry out a variety of dyeing and multiple molecules
The detection of target.Although although centrifuged deposit object smear and directly centrifugation smear can improve target to a certain extent
The quantity and painting tablet quality of cell, but still not can solve cytology film-making limited amount, it is difficult to carry out a variety of dyeing and multiple points
The problem of sub- target detection.Thus, the quality and final diagnosis of cytologic slide have been directly influenced, positive cell is caused
The situation that recall rate is low and false negative rate is high is difficult to improve for a long time.
Therefore, the slice-making quality for improving cytological samples becomes the urgent technical problem that cell pathology faces.Very much
Before year, people have just had recognized that this problem, and propose for body fluid sample to be prepared into cell block (Cell block,
CB imagination and method), to achieve the purpose that the target cell (Karnauchow the P N, et that are enriched in body fluid sample
al.1982.)(Gill G W.2013)。
Currently, cell block technology has become the important technology of processing body fluid sample, advantage also increasingly shows
(Bhanvadia V M,et al.2014；Thunnissen E,et al.2012；Sui Yanxia waits .2017；Xu little Yan, etc.
.2015；Rahbar M,et al.2012；Azami S,et al.2016).Compared with traditional cytology flaking method, carefully
Born of the same parents' wax stone technology overcomes that conventional cell smear cells quantity is few, thickness is different, cell overlap, structure are unclear, can only prepare
A small number of smears, sensibility are low, false negative rate is high, are difficult to the shortcomings that carrying out a variety of dyeing and the detection of multiple molecular targets, can not only
More aim cells are obtained, and the slice eucaryotic cell structure through fixation, dehydration is relatively sharp, improves to the full extent
The slice-making quality of cytological samples and sensibility, specificity, the accuracy of diagnosis, and part has reached histopathology and has examined
Disconnected level becomes a kind of body fluid sample flaking method between traditional exfoliative cytology and histopathology.Especially
The cell block of acquisition can also tens of serial section of sliced acquisition, can be simultaneously using a variety of colouring methods and to a variety of points
The detection of sub- target provides more structurally sound Laboratory evidence for tumor cells diagnosis, Prognosis scoveillance, therapeutic scheme selection.
In addition, cell block can with long-term preservation, help to carry out follow-up and retrospective study to patient.
Currently, cell block technology has become the routine techniques for handling all kinds of body fluid samples, and there is scholar's research to mention successively
Egg white method (Sun Wugang .2017), agar method (Ardengh J, et al.2007), gelfoam out (Guo Xiuyun waits .2013)
(He Hongmin waits .2013 with cell centrifugal pipe process；He Hongmin, CN 103543057 A [P] 2014) etc. various kinds of cell wax stone system
Make method:
(1) for cell component than more rich body fluid sample, a greater number can be obtained using direct centrifugal
Cast-off cells sediment, be added fixer it is directly fixed after, can often make cell precipitation condensation agglomerating, with lens wiping paper or other
Non-fat-soluble water permeable membrane package is put into embedded box, according to the dehydration of conventional organization sample, waxdip, paraffin embedding, can be produced
Cell block (Gu Weina waits .2015).
(2) for the body fluid sample of aim cell negligible amounts, the conventional centrifugal precipitation method can not make cell precipitate solidifying
It clumps together, and is easy to scatter in subsequent operation, satisfactory cell block can not be produced.For this purpose, there is scholar's addition
Egg white (Du Ping waits .2014) or agar gel as cell adhesion matrix, with enhance be aggregated blocking effect (Wang Yongsheng,
Equal .2006), then fixed through fixer, lens wiping paper or other non-fat-soluble water permeable membranes package are put into embedded box, according to routine
Tissue samples dehydration, waxdip, paraffin embedding, can produce cell block.
But merely using egg white or agar not can be well solved cell precipitate condense it is insecure, in subsequent behaviour
It is easy to scatter in work, causes cell block production quality not high or even the problem of failure.Moreover, need to also be using agar gel
Carrying out heating to agar in manufacturing process could melt, not only inconvenient, and heating process is it is also possible to cellular morphology, knot
Structure and function cause irreversible damage, directly affect microscopic observation and the last diagnostic in later period.
Therefore, also someone is centrifuged Manifold technology using cell and " ethyl alcohol -- suction pipe " method (Cheng Kai waits .2017) obtains cell
Precipitating makes cell block after fixation (Wu Han waits .2015).But this method is only capable of assembling cell precipitation, cell is solidifying
It ties and insecure, it is easy to scatter in subsequent operation.
Furthermore there are also the features that scholar utilizes different fixatives, are improved by improvement fixative and set time thin
Born of the same parents' sediment condenses pockets of effect, and (Zhao Jie waits .2013；Rong Hui waits .2014；Hou Fang waits .2017.).For example, there is scholar
Select 10% Ethanol-Acetic Acid acid formaldehyde formulations, AAF (95% 4 milliliters of 2 milliliters+formalin of ethyl alcohol 34ml+ glacial acetic acid), 9%
The formaldehyde of ethyl alcohol+40%, 100% ethyl alcohol and 10% formaldehyde equivalent mixed liquor, Bouin solution (acid+glacial acetic acid of saturation+
Formalin) it is fixed as fixative to make cell block, or with formalin or IPA vapor
(Frederick M, et al.2010) at least fixes 2 hours (Vinayakamurthy S, et al.2016).
Although traditional cell block preparation method can be prepared by corresponding cell block.But it can not be fundamentally
Insecure, to be easy to scatter in subsequent operation critical issue is adhered between cell precipitate inner cell after upper solution centrifugation.This
Outside, the additives such as agarose need to also be heated in operating process, can also generate not expected adverse effect to sample.
Therefore, grope new cell block preparation method, do not influence cellular morphology, function and the inspection of subsequent molecular target
Under the premise of survey, enhancing cell precipitate condenses pockets of fastness, is that current cell block prepares the critical issue faced,
And there is important clinical medical value and economic significance.
Summary of the invention
In order to overcome the disadvantages of the above prior art, the purpose of the present invention is to provide a kind of sizings of cell block to fix
Plate and the method for making cell block with it, the cell block are formed fixed plate structure design rationally, easy to use, Neng Goushi
The good plastotype of existing cell precipitate is fixed；The cell block production method is easy to operate, cell precipitate can be made with approximation
Regular form is embedded in paraffin mass.
In order to achieve the above object, the present invention is achieved by the following scheme:
The invention discloses a kind of cell block sizing fixed plates, including sizing fixed plate, open up in sizing fixed plate
If there is dry setting fixation hole；
Sizing fixation hole is cylindrical hole, and bottom is smooth or bottom is U-shaped, for containing cell precipitate；
The diameter of sizing fixation hole, hole depth is adjustable.
Preferably, the sizing fixation hole is in array distribution in sizing fixed plate.
Preferably, the diameter of sizing fixation hole (1) is 3~10mm, and depth is 5~15mm.
The invention also discloses a kind of production method of cell block, this method is solid based on above-mentioned cell block sizing
Fixed board is realized, comprising the following steps:
1) Sample pretreatment
Sample liquid to be prepared is stood, cell precipitation is obtained, centrifugal treating goes supernatant to obtain cell precipitate,
The cell precipitate is transferred in the sizing fixation hole opened up in sizing fixed plate；
2) thickener is added
According to the volume for the cell precipitate that step 1) obtains, according to the volume ratio of 1:0.5~1, to cell precipitate institute
Sizing fixation hole in be added water-based thickener protein solution, mix well；
3) cell block is made
95% alcohol is added into the sizing fixation hole added with water-based thickener protein solution, stewing process, until
It is agglomerating to observe cell precipitate condensation, is then wrapped up with lens wiping paper or non-fat-soluble water permeable membrane, is placed in embedded box and is fixed
Finally cell block is made through dehydration, the processing of transparent, waxdip, paraffin embedding in the cell precipitate sample fixed by processing.
Preferably, in step 2), water-based thickener protein solution is that 0.2~5g water-based thickener and 5~10g is auxiliary
It helps albumen to be dissolved in 100mL PBS buffer solution to be formulated.
Preferably, the water-based thickener is xanthan gum, algin sodium, pectin, algin propylene diester, carboxymethyl
One or more of cellulose, gum arabic and guar gum.
Preferably, the auxilin uses ovalbumin or bovine serum albumin(BSA).
Preferably, according to the volume of the cell precipitate containing water-based thickener protein solution, according to 1:5~10
Volume ratio is slowly added to 95% alcohol, stands 10~15 minutes and is pre-fixed.
Preferably, in step 3), the fixing process 4~24 in the stationary cylinder for filling 10% neutral formalin fixer
Hour.
It preferably, further include that the cell block sections that will be obtained handle the operation that paraffin section is made；
Gained paraffin section, for observing eucaryotic cell structure and detection specific molecular target, or for extracting albumen, core
Acids molecule is detected accordingly.
Compared with prior art, the invention has the following advantages:
Sizing fixed plate provided by the invention may be implemented to fix the good plastotype of cell precipitate, and the utensil
Structure is simple, easy to use, and experimental result confirms, the cell block prepared using method provided by the invention and utensil, cell
Sediment condensation is close, and spallation, broken phenomenon are less prone in subsequent operation.
Cell block production method provided by the invention is simple, enables cell precipitate quilt in the form of substantially regular
It is embedded in paraffin mass.Meanwhile present invention utilizes the good suspended solid materials of water-based thickener, increase solution viscosity
Characteristic, and to temperature, acid, alkali, salt, oxidant and protease stablize, it is soluble easily in water and do not dissolve in alcohol the characteristics of, by it
It is configured to thickener protein solution, and using alcohol as agent is pre-fixed, has well solved conventional cell wax stone preparation method
Middle cell precipitate condensation effect is bad, is easy the fragmentation in subsequent operation, leads to the shortcomings that cell block production failure.
The present invention is by specific it is demonstrated experimentally that the cell block of this method preparation is to eucaryotic cell structure and subsequent molecular target
Mark detection has no effect, and nor affects on detection of the subsequent experimental to molecular target.Moreover, this method is easy to operate, at low cost
Paraffin section honest and clean, that thus prepared by cell block, after HE is dyed, cellular morphology is complete, subcellular structure is clear, is easy to sentence
It reads, immunohistochemical staining and in situ hybridization testing result also can get satisfied testing result.Therefore, the present invention mentions
Crucial technical problem in the existing cell block preparation method of the method and utensil very good solution of confession, can be in certain journey
The level for reaching histopathology slide on degree fully meets the needs of cell pathology diagnosis and molecular target detection.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of cell block sizing fixed plate of the invention；
Fig. 2 is cell block sizing fixed plate the schematic diagram of the section structure of the invention；
Fig. 3 is through the fixed cell precipitate coagulated agglomerate photo being put into embedded box；
Fig. 4 is the cell block prepared and corresponding paraffin section；
Fig. 5 is the cell mass integrality comparison result photo made of the method for the present invention and conventional method；
Fig. 6 is the cell block integrality comparison result photo made of the method for the present invention and conventional method；
Fig. 7 is the integrality comparison result photo of the cell block sections made of the method for the present invention and conventional method；
Fig. 8 is the expression of results of CA125 in the cell block for compare distinct methods production using ImmunohistochemistryMethods Methods.
Wherein: 1- sizing fixed plate；The qualitative fixation hole of 2-.
Specific embodiment
In order to enable those skilled in the art to better understand the solution of the present invention, below in conjunction in the embodiment of the present invention
Attached drawing, technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is only
It is the embodiment of a part of the invention, instead of all the embodiments.Based on the embodiments of the present invention, the common skill in this field
Art personnel every other embodiment obtained without making creative work, all should belong to protection of the present invention
Range.
It should be noted that description and claims of this specification and term " first " in above-mentioned attached drawing, "
Two " etc. be to be used to distinguish similar objects, without being used to describe a particular order or precedence order.It should be understood that making in this way
Data are interchangeable under appropriate circumstances, so that the embodiment of the present invention described herein can be in addition to scheming herein
Sequence other than those of showing or describe is implemented.In addition, term " includes " and " having " and their any deformation, it is intended that
Be to cover it is non-exclusive include, for example, containing the process, method, system, product or equipment of a series of steps or units
Those of be not necessarily limited to be clearly listed step or unit, but may include be not clearly listed or for these processes,
The intrinsic other step or units of method, product or equipment.
The invention will be described in further detail with reference to the accompanying drawing:
1, cell block manufacturing appliance
It is a kind of cell block manufacturing appliance of the invention, i.e. cell block fixed plate referring to Fig. 1, including sizing is fixed
Plate 1, if being densely covered with dry setting fixation hole 2 in sizing fixed plate 1, the sizing fixation hole 2 is cylindrical hole, and bottom is smooth.
Or sizing fixation hole 2 is U-shaped, the diameter and depth for fixation hole 2 of being formed are adjustable, adjust size according to actual needs.
It is recommended that the diameter of sizing fixation hole 2 is 3~10mm, depth is 5~15mm.
The cell block fixed plate can be according to the difference of material, using the processing of the techniques such as injection molding, blow molding or drilling
At.
2, main agents and material
Prepare the thickener protein solution for containing appropriate thickener and auxilin.
Wherein, used thickener can be xanthan gum, algin sodium, pectin, algin propylene diester, carboxylic first
One of base cellulose, gum arabic and guar gum are a variety of, preferably xanthan gum.
Used auxilin is ovalbumin or bovine serum albumin(BSA).
Thickener protein solution is prepared with the following method: first being dissolved auxilin using PBS buffer solution, is configured to
Final concentration of 5%~10% auxilin solution, then thickener albumen is configured to auxilin solution dissolution thickener
Aqueous solution, thickener final concentration of 0.1%~2.0% in thickener protein solution.
3, the production method and process of cell block
(1) pre-treatment of body fluid sample
According to the volume of the various body fluid samples such as the Pleural effusions of inspection, urine, the centrifuge tube of suitable size and corresponding is selected
Cytospin is centrifuged 10~15min with 1000~3000r/min, after abandoning supernatant, sediment containing cell is obtained, by this
Sediment stays in centrifuge tube or it is suitably formed fixation hole 2 with the size that suction pipe is transferred in sizing fixed plate 1
It is interior.
(2) addition of thickener protein solution
According to the volume of sediment in centrifuge tube or sizing fixation hole, and according to the volume ratio of sediment (1:0.5~1)
Above-mentioned prepared thickener protein solution is added and mixes.
(3) fixation of sample
1) it pre-fixes: according to the volume of sediment to be fixed in centrifuge tube or sizing fixation hole, according to the body of 1:5~10
Product than being slowly added to 95% alcohol along sizing fixation hole inward flange, fix 10~15 minutes and condense into cell precipitate by standing
Group.
2) fixed: pockets of cell precipitate lens wiping paper will to be condensed in centrifuge tube or sizing fixation hole or other are non-fat
It is placed into after dissolubility water permeable membrane package in conventional disposable biopsy specimen embedded box with cover, and record is numbered, then
It puts it into and fills enough 10% neutral formalin fixers and (be according to the volume of fixation and the volume ratio of fixer
1:5~10) fixation container in, fix 4~24 hours, as a result as shown in Figure 3.
(4) it is dehydrated and embeds
It will be handled through well fixed Cell clumping object according to routine autopsy tissue dewatering, waxdip, transparent, embedding program,
Carry out paraffin embedding, can acquisition cell block processed, then through conventional slicing treatment, the cell section of acquisition, as a result such as Fig. 4.
4, the integrality comparative experiments of the method for the present invention and the cell block of conventional method production
Select the chest and abdomen water of inspection to be greater than 35, sample of 100ml or more, at the same using the method for the present invention and tradition from
The heart precipitation method make cell block respectively, and be sliced from the cell mass, cell block, HE of production general form, eucaryotic cell structure
And detection albumen the methods of subcellular localization be compared, analyze the method for the present invention the characteristics of.
Specific experiment condition and method are as follows: equivalent (50ml) chest and abdomen water sample first being taken to pour into two cell centrifugations respectively
Supernatant is abandoned after being centrifuged (2000 revs/min) in pipe 15 minutes, wherein a sample the method according to the invention sinks this
Starch, which continues to employ the size that suction pipe is transferred in sizing fixed plate 1 and is suitably formed in fixation hole 2, sequentially adds isometric thickening
Agent protein solution simultaneously mixes, and is then pre-fixed 10 minutes with 95% alcohol standing, it is solid to add enough 10% neutral formalins
Determine liquid and fixes 4 hours；Enough 10% neutral formal directly is added along centrifugation tube wall according to conventional centrifugal method in another sample
Woods fixer fixes 4 hours.Then the cell mass after two methods are fixed is carefully transferred to corresponding number with tweezers respectively
Embedded box in be dehydrated, waxdip, transparent, embedding, can acquisition cell block processed, then through conventional paraffin section and HE
Dyeing, the cell section of acquisition.Expression using Immunohistochemical Method detection CA125 in chest and abdomen water sample simultaneously.As a result such as
Fig. 4.
Integrality is relatively mainly reflected in cell mass, cell block, HE slice and detection protein expression etc., knot
Fruit shows as follows:
1) cell mass
As shown in figure 5, A-J in Fig. 5: using the cell mass of method provided by the invention preparation, K-L: using conventional centrifugal
The precipitation method production cell mass, it can be seen that using the conventional centrifugal precipitation method make cell mass it is loose, it is difficult to condense it is agglomerating,
And use the bonding of cell mass made from method of the invention close.
2) cell block
As shown in fig. 6, A-D in Fig. 6: the cell block of conventional centrifugal intermediate processing production；E-H: it is provided using the present invention
Method preparation cell block, it can be seen that cell mass made from the conventional centrifugal precipitation method is loose, and use present invention side
The bonding of cell mass made from method is close, structural integrity.
3) HE is sliced
As shown in fig. 7, A in Fig. 7: B: the HE slice of the cell block of conventional centrifugal precipitation method production is mentioned using the invention
The cell block of the method preparation of confession, it can be seen that the HE slice cell of conventional centrifugal precipitation method preparation is loose, it is difficult to be formed
Complete structure；And use HE prepared by the present invention slice display cell mass bonding close, structural integrity.
4) expression of CA125
As shown in figure 8, A in Fig. 8: the cell block of conventional centrifugal precipitation method production, CA125 protein expression in cytoplasm,
But eucaryotic cell structure is unintelligible, and cell membrane is imperfect.B uses the cell block of method provided by the invention preparation, CA125 albumen
It is expressed in cytoplasm, eucaryotic cell structure is clear, and cell membrane is complete.
5. the application of cell block
The cell block prepared according to the method provided by the invention is sliced using paraffin slicing machine, can be obtained
Required paraffin section is obtained, which can be used for HE dyeing, specific stain and use immunohistochemistry, original position miscellaneous
The in situ tissues cytologies such as friendship detect albumen, nucleic acid molecular target.
In conclusion the present invention devises dedicated for carrying out the fixed sizing fixed plate of moulding to cell precipitate, and
It is configured to using the biological characteristics of water-based thickener, especially xanthan gum, and by it with ovalbumin or bovine serum albumin(BSA)
Water-based thickener protein solution, with it is isometric than be added to mixed in cell precipitate after using 95% alcohol as pre-fixing
Agent is pre-fixed, so that cell precipitate condensation is agglomerating, then 10% neutral formalin is used to be fixed as fixer,
Most it can be obtained cell block through dehydration, waxdip, embedding afterwards.This method fundamentally solves conventional cell wax stone preparation side
The shortcomings that method keeps cell precipitate condensation bad, is easy to spallation in subsequent operation.Meanwhile the thickeners such as xanthan gum used
With auxilin in cell block preparation will not form to cell and result generate any adverse effect, meanwhile, also not shadow
Ring detection of the subsequent experimental to molecular target.
The above content is merely illustrative of the invention's technical idea, and this does not limit the scope of protection of the present invention, all to press
According to technical idea proposed by the present invention, any changes made on the basis of the technical scheme each falls within claims of the present invention
Protection scope within.
Claims (10)
- The fixed plate 1. a kind of cell block is formed, which is characterized in that including being formed fixed plate (1), opened in sizing fixed plate (1) Equipped with dry setting fixation hole if (2)；Fixation hole (2) are formed as cylindrical hole, bottom is smooth or bottom is U-shaped, for containing cell precipitate；The diameter of sizing fixation hole (2), hole depth is adjustable.
- The fixed plate 2. cell block according to claim 1 is formed, which is characterized in that the sizing fixation hole (2) is fixed It is in array distribution in type fixed plate (1).
- The fixed plate 3. cell block according to claim 1 is formed, which is characterized in that the straight of fixation hole (1) of being formed Diameter is 3~10mm, and depth is 5~15mm.
- 4. a kind of production method of cell block, which is characterized in that this method is based on described in any one of claims 1 to 3 Cell block sizing fixed plate realize, comprising the following steps:1) Sample pretreatmentSample liquid to be prepared is stood, cell precipitation is obtained, centrifugal treating goes supernatant to obtain cell precipitate, by this Cell precipitate is transferred in the sizing fixation hole (2) opened up in sizing fixed plate (1)；2) thickener is addedAccording to the volume for the cell precipitate that step 1) obtains, according to the volume ratio of 1:0.5~1, to where cell precipitate It is formed in fixation hole (2) and water-based thickener protein solution is added, mix well；3) cell block is made95% alcohol, stewing process are added into the sizing fixation hole (2) added with water-based thickener protein solution, until seeing It is agglomerating to examine cell precipitate condensation, is then wrapped up with lens wiping paper or non-fat-soluble water permeable membrane, is placed in embedded box and place is fixed Finally cell block is made through dehydration, the processing of transparent, waxdip, paraffin embedding in the cell precipitate sample fixed by reason.
- 5. the production method of cell block according to claim 4, which is characterized in that in step 2), water-based thickener egg White water solution is that 0.2~5g water-based thickener and 5~10g auxilin are dissolved in 100mL PBS buffer solution to be formulated.
- 6. the production method of cell block according to claim 4, which is characterized in that the water-based thickener is xanthan One of glue, algin sodium, pectin, algin propylene diester, carboxymethyl cellulose, gum arabic and guar gum or It is several.
- 7. the production method of cell block according to claim 4, which is characterized in that the auxilin uses egg white egg White or bovine serum albumin(BSA).
- 8. the production method of cell block according to claim 4, which is characterized in that in step 3), according to containing aqueous The volume of the cell precipitate of thickener protein solution is slowly added to 95% alcohol according to the volume ratio of 1:5~10, stands It is pre-fixed within 10~15 minutes.
- 9. the production method of cell block according to claim 4, which is characterized in that in step 3), fill 10% neutrality Fixing process 4~24 hours in the stationary cylinder of formalin fixer.
- 10. the production method of cell block according to claim 4, which is characterized in that further include the cell wax that will be obtained The operation of paraffin section is made in block slicing treatment；Gained paraffin section, for observing eucaryotic cell structure and detection specific molecular target, or for extracting albumen, nucleic acid point Son is detected accordingly.
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| CN117660604A (en) * | 2024-02-01 | 2024-03-08 | 广州迈景基因医学科技有限公司 | FFPE reference for NGS detection and preparation method and application thereof |
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| CN102719352A (en) * | 2012-06-06 | 2012-10-10 | 西安交通大学 | Cell chip slide for preparing microarray cell chips and preparation method |
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| CN102719352A (en) * | 2012-06-06 | 2012-10-10 | 西安交通大学 | Cell chip slide for preparing microarray cell chips and preparation method |
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| CN117660604A (en) * | 2024-02-01 | 2024-03-08 | 广州迈景基因医学科技有限公司 | FFPE reference for NGS detection and preparation method and application thereof |
| CN117660604B (en) * | 2024-02-01 | 2024-04-16 | 广州迈景基因医学科技有限公司 | FFPE reference for NGS detection and preparation method and application thereof |
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Application publication date: 20191105 |