CN101629946B - Urinary cell micro staining analysis method - Google Patents
Urinary cell micro staining analysis method Download PDFInfo
- Publication number
- CN101629946B CN101629946B CN200810040597.7A CN200810040597A CN101629946B CN 101629946 B CN101629946 B CN 101629946B CN 200810040597 A CN200810040597 A CN 200810040597A CN 101629946 B CN101629946 B CN 101629946B
- Authority
- CN
- China
- Prior art keywords
- cell
- urine
- dyeing
- staining
- micro
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to the field of medical tests, and relates to a urinary cell micro staining analysis system, in particular to a urinary cell micro staining analysis method and an involved analyzing device. Combining with a computer automatic control and recognition technique, the invention provides a urinary cell micro staining quantitative system which can achieve cell separation, extraction and thin layer flaking of arena, support a single-flow micro staining technique, and aim at clinical large samples and rapid analysis. The system overcomes the defects of the prior art, and the results of urine sample clinical medical inspection practices show that the urinary cells are stained clearly, are distributed evenly and are easy to recognize and quantitate. The method can prevent sample cross contaminations and can perform precise quantitative analysis without staining solution environmental pollution. The method is in accordance with the requirements on arena examination standardization established by the inspection branches of International (NCCL) and Chinese Medical Association.
Description
Technical field
The invention belongs to field of medical examination, relate to urinary cell micro staining analysis.Be specifically related to support micro-staining technique a kind of comprising, prevent sample cross contamination, the urine sediment analysis method that accurate quantification is analyzed.
Technical background
Clinical examination practice shows, arena complicated often needs to adopt relevant analytical equipment to carry out the detection analysis of arena.So far, the sediment urinalysis analyser that there is not yet prior art is carried out the each component in arena is distinguished to extraction process analysis.While dyeing due to these existing analysers, cannot avoid the cross pollution that influences each other, do not support that trace dyeing and thin layer film-making, dyeing error are large, has sample of arena composition, false positive rate is high, often need manual operations to cause poor repeatability, length consuming time, and cannot carry out accurate quantification counting and dyeing condition cannot standardization.Only specific demand patient, just carry out the chromoscopy of arena, cannot meet large-scale arena staining examine clinically.Therefore the detection of Clinical Urinary sediment is at present analyzed and mainly rested in non-dyeing, qualitative detection level, the method for there is no is carried out accurate quantification analysis.Due to kidney cell clinically come off and the clinical meaning of leukocyte infiltration has important value for activity and the prognosis of judging kidney trouble, yet home and abroad there is no the quantitative analysis tech device for urinary cell micro dyeing at present.
The Varistain Gemini pathology automatic slide stainer of Britain Thermo Shandon company development, has realized the robotization of tissue section strain, also can be used for urinating cell dyeing.But there is following defect: single casing design cannot be avoided the cross pollution of urine sample; Can not implement trace and control the dosage of dye liquor; Dyeing, for completing in batches, only can be supported a kind of dyeing simultaneously, cannot realize the personalization dyeing of single sample; In operation, need often to change dye liquor and clean the waste that dye liquor cylinder causes a large amount of dye liquors; Without automatic ration counting procedure, without the treating apparatus of waste water and toxic gas, therefore this machine is only suitable for histopathologic slide's dyeing of small sample.
Summary of the invention:
The object of the invention is, for overcoming the defect of prior art, provides a kind of urinary cell micro staining analysis system.Relate in particular to a kind of urinary cell micro staining analysis method and related analytical equipment thereof.
The present invention in conjunction with computer controlled automatic and recognition technology provide can for clinically, large sample, express-analysis be object urinary cell micro dyeing quantitative system, this system can realize cell separation, extraction, the thin layer film-making of arena, and support single process trace staining technique, prevent sample cross contamination and accurate quantification analysis and without dye liquor environmental pollution.
The present invention meets the standardized requirement of uropsammus examination of the world (NCCL) and the formulation of check branch of Chinese Medical Association.
Object of the present invention is achieved through the following technical solutions:
1, a kind of method of urinary cell micro staining analysis is provided, and described method comprises the steps:
1) after urine sample separation to be measured, urinate cell sample,
Urine sample to be measured enters urine cell gradient density centrifugation sampling apparatus, after the automatic separation of urine sample cell separator, proceeds to urine cell sample device, through density gradient centrifugation collection urine cell liquid;
2) urine sectioning cells,
The urine cell liquid gathering, after dilution, moves to rejection tablet film-making automatically in automatic pelletizer,
3) urine cell sheet dyes sheet,
The urine cell sheet making moves into cell automatic fine dyeing apparatus and dye sheet under the software control of slide Dyeing control;
4) gather urine cell and dye picture
Urine cell dyes sheet and inserts high-resolution color numeral micrometron platform, through micro-platform automatically, controls software control, through microscope, video high Definition CCD image acquisition device, is sent on LCD colorful display screen to obtain image,
5) urine cell classification mark and image photographic, qualitative and quantitative analysis,
Step 4) urine cell dyes picture and urinate cell classification mark and image photographic behind the urine cell image Real-time Collection automatic analysis selected staining cell of quantitative Treatment software region, automatically carries out qualitative and quantitative analysis;
6) print urine test result picture and text report, grouping of data management
Above-mentioned analysis result is controlled and is carried out colour print through urine test project picture and text report print software, and patient medical record clinical data carries out Classification Management through graphic-text database management software.
After the treated device of above-mentioned dyeing liquor and waste gas purifies, discharge, in waste gas wherein, benzene concentration exceeds standard and controls automatic alarm by alarm software.
2, provide the described related analytical equipment of urinary cell micro staining analysis system,
Described analytical equipment comprises urine sample cell separation apparatus, urine cell gradient density centrifugation sampling apparatus, automatically spreads sheet devices, automatic staining device and dyeing liquor treating apparatus, high Definition CCD image acquisition device, high-resolution color numeral micrometron platform and microscope, computer control system and control software, LCD colorful display screen and color printer (Fig. 1).
Described cell separation apparatus (Fig. 2) is by filter screen irrigator [2], and filter screen [3] and screen support point [5] form, and wherein filter screen is 30～300 orders; When freshly voided urine process filter screen irrigator, the compositions such as the separated arena cast of energy, cellulose;
Described urine cell gradient density centrifugation sampling apparatus (Fig. 3) is by urine cellular collector [1], and switch [5] rubber stopper [6] air metal catheter [7] automatic control flow [8] is controlled plug [B] and formed, and the Ficoll liquid [4] of variable concentrations is housed in device;
After urine sample separation, enter described urine cell gradient density centrifugation sampling apparatus, the compositions such as urine cell of crossing net slowly inject arena Hanks liquid and in Ficoll, are made into the gradient of continuous density test tube of 1.300～1.010g/ml under apocarpy, are controlled the cell mass of collecting layering by the air metal catheter on rubber stopper;
Described automatic pelletizer (Fig. 4,5): by [1] base; [2] support; [3] acellular high density Ficoll waste liquid; [4] disposable plastic cell rejection tablet device; [5] rejection tablet device stationary installation; [6] circular rubber band; [7] microslide spiral fixator; [8] microslide; [9] exhausted bath box; [10] switch; [11] high density Ficoll urine cell mixture; [12] waste liquid flows out pipeline formation;
After the cell mass of above-mentioned collection layering is centrifugal, with Hanks liquid, suspend, according to UVA value, adopt Hanks liquid to carry out sample dilution, after getting a certain amount of urine cell sample and adding and mix in higher density solution, move in cell rejection tablet pipe, by centrifugal several minutes of described pelletizer, the cell in liquid base is got rid of automatically in microslide cell hole made thin-layer cell, liquid flows into exhausted bath box through filter paper, fixing dry rear standby with absolute ethyl alcohol;
Described automatic staining device (Fig. 6): by 1. admission gears, 2. air groove 3. dye liquor tank 4. microslide rotating disks, 5. control panel 6. motors, 7. water mortise, 8. micro-stain pipettor, 9. air exhausting device 10. filtration units, 11. woven hoses, 12. Rinsing Areas form;
During dyeing, multi-disc thin-layer cell microslide is placed on to dying in film trap (Fig. 7) of automatic staining device fixing, completely separated between staining section, under controlling, micro-Dyeing control software systems input respectively the dyeing course of each position.In above-mentioned slide trace Dyeing control software systems, determined length of a game's axle (calling GTL in the following text) of a dynamic management, and introduced the probabilistic search module based on genetic algorithm idea;
In length of a game's axle of described dynamic management, the list structure that single dyeing action is minimum treat node is take in employing, in logic mark each time etching system dyeing behavior, and the interface that provides insertion, inquiry etc. to operate by one group of function, in order to guarantee that each action occurred in the correct moment in whole dyeing course, guarantee that any action can be because resource contention produces conflict, not guarantee the coordination of each action and efficient.
During operation, described GTL for empty, along with entering the generation of the events such as sheet, dyeing course definition, slice, coordinates pending action by control module when system starts to put into operation, and after determining true generation constantly, generates node in GTL; This node is described the action that will occur in moment, the position of main turntable, the time taking and this period is occurred, and provides foundation for the scheduling result of control module after current time.Table 1 is GTL organization definition.
Table 1
| Typedef unsigned long timeFlag; //max:4294967295; Typedef unsigned short timeMeasure in 136.2; //max:65535; 18.2 hours typedef bitset<MAXfunPlace>II_int; The axle node */unsigned int index of typedef struct GTL{/* length of a game; // node index timeFlag tf; // the time is carried timeMeasure tc; The time period size timeMeasure circle that // node is described; // be the reserved rotation time II_int postionBIT of operation of this piece; // list of locations is with respect to No. 0 position struct GTL * prep; Struct GTL * next; TimeLine, * TimeLineLink; |
The described probabilistic search module (calling NescafeGA in the following text) based on genetic algorithm idea, on the idea basis of GTL, construct appropriate dispatching sequence, so that operating in efficiently, dyeing subsystem can in solution space, search for optimum solution, thereby raising running efficiency of system, under the prerequisite of precision of guaranteeing to dye, efficient processing dyed sheet.
In this NescafeGA, adopt while the technical approach of seeking strategy, implementation strategy, the processing power of solution embedded system limited with the harsh requirement of user to system response time.
Above-mentioned micro-Dyeing control software systems of the present invention can effectively be utilized the dyeing stand-by period that will inevitably occur in the quantitative staining analysis process of arena, and, in NescafeGA operational process, monitoring processor load constantly, and to regulate dynamically its CPU usage.The described relevant informations such as NescafeGA reading system process processing time, and determine thus the CPU usage of NescafeGA.By system, sleep calls, and when system need be processed high priority action, as processed GUI (graphic user interface), coloration result etc., abdicates cpu resource.
Data definition in above-mentioned list structure is as following, take 24 dye sheet as example:
int allele[24]＝{1，21，6，12，15，10，20，22，13，4，18，11，5，2，8，23，16，0，14，19，7，3，17，9}
typedef struct alleleST{
char arrange[CardNum]
}allele，*allelelink；
typedef struct chromatidST{
unsingned int fitness；
unsingned int result；
double probability；
allelelink head；
}chromatidST，*chromatidlink；
initPOP()；
initOutstandingPool()；
while(Generation＜MaxGeneration && now-start＜CalculateTime && Minmodify)
{
CalculateFitness()；
getnexFitness()；
Generation++；
The chromatid number of a population can be specified by parameter, and monomer fitness is determined by the scheduling sequence complete operation required time of describing according to this chromosome.And evolve with the variation probability of 0.6 crossover probability and 0.2.Evolution termination condition is determined by 3 factors: 1. reach the maximum algebraically of evolving, 2. calculate and T.T. surpass predetermined value, 3. processing time rate of change corresponding to current optimum solution is less than predetermined value.
The present invention provides the specific implementation process of control module:
1) natural language of HE dyeing course is:
1s haematine dye liquor; Wait for 210s; 210s tap water; 1s 0.5% hydrochloride alcohol; Wait for 7s; 210s tap water; 1s 0.5% ammoniacal liquor oil blackeite; Wait for 45s; 210s tap water; 210s distilled water; 1s 0.5% Yi Hongshui; Wait for 180s; 15s distilled water; 23s 80% alcohol; 23s 95% alcohol; 90s 100% anhydrous alcohol; 1s dimethylbenzene; Wait for 90s; 1s dimethylbenzene; Wait for 240s; 2s neutral gum.
2) to translate above-mentioned natural language be following sequence to control module, take 1 as example,
F2_L2_S1.##__210^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F10_L4_S1.##__7^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F11_L4_S1.##__45^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F30_L20_L21_S210.F12_L5_S1.##__180^0.F30_L20_L21_S15.F13_L9_L10_L11_S23.F5_L9_L10_L11_S23.F7_L9_L10_L11_S90.F8_L3_S1.##__90^30.F8_L3_S1.##__240^60.F9_L4_S2
Event sheet is separated by point (.); The event sheet starting with alphabetical F, represents to need that system respond with stain or rinse and move; The event sheet that the character # of take starts is waiting event, does not need system to provide action.As: F2_L2_S1.##__210^0.F5_L9_L10_L11_S23
Wherein, haematine dye liquor is defined as No. 2 functions (F2) and is assemblied on No. 2 positions (L2) this event effect 1 second (S1); Waiting event starts (##__) with two No. # and two underscores and adds the stand-by period, and ' ^ ' number represents that heel numeral waits for error of fragment permission for this, so ##__210^0 represents to wait for 210 seconds, and error should be controlled at ± in 0 second; With 80% alcohol, rinse 23 seconds, be expressed as F5_L9_L10_L11_S23, wherein 80% alcohol functionality number 5 (F5), are arranged on 9,10, No. 11 positions (_ L9_L10_L11), act on 23 seconds (S23).
It is following sequence that control module is translated above-mentioned multi-disc natural language,
In clobal table descriptive system, the needed dyeing course of institute's sheet of having illicit sexual relations of current loading, is comprised of multirow, and line number is the maximum load amount of system.
0！；
1：F2_L2_S1.##__210^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F10_L4_S1.##__7^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F11_L4_S1.##__45^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F30_L20_L21_S210.F12_L5_S1.##__180^0.F30_L20_L21_S15.F13_L9_L10_L11_S23.F5_L9_L10_L11_S23.F7_L9_L10_L11_S90.F8_L3_S1.##__90^30.F8_L3_S1.##__240^60.F9_L4_S2；
2：F2_L2_S1.##__210^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F10_L4_S1.##__7^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F11_L4_S1.##__45^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F30_L20_L21_S210.F12_L5_S1.##__180^0.F30_L20_L21_S15.F13_L9_L10_L11_S23.F5_L9_L10_L11_S23.F7_L9_L10_L11_S90.F8_L3_S1.##__90^30.F8_L3_S1.##__240^60.F9_L4_S2；
3：F30_L20_L21_S5.F14_L5_S1.##__1200^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S60.F15_L5_S1.##__15^5.F29_L12_L13_L14_L15_L16_L17_L18_L19_S90.F30_L20_L21_S7.F16_L6_S1.##__240^60.F29_L12_L13_L14_L15_L16_L17_L18_L19_S90.F30_L20_L21_S7.F17_L6_S1.##__60^0.F18_L6_S7.F9_L4_S2；
4！；
5！；
6！；
7！；
8！；
24！
Above-mentioned sequence description the situation of system loading multi-disc while dying sheet.Line number opens from zero, and every row is started by line number, heel dyeing course descriptor, and with branch (; ) ending.If certain row unloaded is dyed sheet, by exclamation mark (! ) replacement dyeing course descriptor.In upper table, first trip " 0! ; " representing that No. 0 position of rotating disk do not dye sheet, the second No. 1, line display position has been loaded and has been dyed sheet, lists file names with the dyeing course that this dyes sheet requirement, and same, 2,3 positions have also been loaded and have been dyed sheet.Here only for describing the problem, unlisted whole length of a game shows.In fact, clobal table can be comprised of the line number of the maximum load amount of system (24).Clobal table is the input of control module algorithm.
The output sequence form of control module is as follows:
Begin ###
C：0 BP：1
S TF0 L2 Fn2
R TF1 L2 Fn2
C：1 BP：0
S TF3 L2 Fn2
R TF4 L2 Fn2
C：9 BP：14
S TF14 L20 Fn30
R TF19 L20 Fn30
C：6 BP：11
S TF213 L12 Fn29
S TF216 L13 Fn29
R TF423 L12 Fn29
R TF426 L13 Fn29
End ###
In output sequence, concrete response action of every line display.Specifically be divided into three kinds: C character starts, represent master rotation.S character starts, and presentation function is opened.R character starts, and presentation function is closed.In " C:9 BP:14 ", 9 expression systems are the reserved rotational time of this rotation process.14 represent take that No. 0 Ran Pian position is as with reference to master is turned to position 14.Sequence is as the input of system dyeing action processing module, and is finally converted into mechanical action.
Above-mentioned dyeing completes automatically, does not interfere with each other, without cross-infection; Described device adopts airtight design, and under its circular staining trough, for sky sets up meter, dyeing liquor all flows into the bottom efferent tract of dyeing apparatus, through adsorbent equipment, enters sewer; Gas is after the top absorption of machine outside discharge chamber (Fig. 6,7); Thin-layer cell microslide mounting is preserved; After purifying, the treated device of dyeing liquor and waste gas discharges; As benzene concentration in waste gas exceeds standard alarm software by automatic alarm;
The present invention dyes sheet analysis by following method to above-mentioned urine cell:
With counting scale microslide, carry out computer picture scale calibration rear (Fig. 8), with automatic micro-platform, control software by the thin layer urine cell microslide picture position under mouse control microscope, by high Definition CCD image acquisition device LCD colorful display screen observe image and under urine cell image auto Analysis is controlled capturing digital image, count tag target cell and area, automatically calculate every microlitre target cell number, urinate cell classification mark and image photographic, automatically carry out qualitative and quantitative analysis.Its result is controlled and is carried out colour print through urine test project picture and text report print software, and patient medical record clinical data carries out Classification Management through graphic-text database management software.
Urinary cell micro staining analysis system of the present invention is put into practice through urine specimen clinical medical inspection, and result shows: urine cell dyeing is clear, is evenly distributed, and is easy to identification and quantitative.Can for clinically, the urinary cell micro dyeing quantitative test that is object of large sample, express-analysis, can realize cell separation, extraction, the thin layer film-making of arena, and support single process trace staining technique, prevent sample cross contamination and accurate quantification analysis and without dye liquor environmental pollution.
The present invention meets the standardized requirement of uropsammus examination of the world (NCCL) and the formulation of check branch of Chinese Medical Association.
The present invention has advantages of following outstanding:
1) carry out single process automatic staining, dyeing kind, dosage and the dyeing time of various dyeing liquors are accurately controlled, thereby realized personalized dyeing;
2) due to the design of single process automatic staining, centrifuge tube and rejection tablet pipe can high temperature, autoclave sterilization can prevent cross pollution completely;
3) support micro-staining technique, can support to carry out quantitative immunohistochemical staining and data analysis, support the ordinary stain of histopathologic slide simultaneously;
4) urine cell separation and density gradient centrifugation design, avoid arena composition influencing each other to dyeing; Thin layer higher density fluid basal cell Wholemount, makes cellular morphology complete, clear in structure; Scale is proofreaied and correct; Area of computer aided quantitative test;
Guaranteed accurate quantification analysis.Thereby the urine cell automatic staining system that provides a kind of uropsammus examination standardization that meets the world (NCCL) and the formulation of check branch of Chinese Medical Association to require;
5) precise time is measured, and multiple colouring method can carry out simultaneously, and clinical samples, without wait, can dye in time, saves time, laborsaving;
6) support micro-staining technique, avoid a large amount of consumption of natural resource, in airtight space, work, and be furnished with char filter, can effectively prevent that chemical reagent, dyestuff are to the pollution of working environment and the harm to human body;
7) can on screen, realize monitoring in real time, control the quality of dyeing;
8) menu mode design, simple to operate, can dye in batches, and dyeing course is precisely controlled, and effect is good, and comparability is strong, is particularly useful for the batch dyeing that workload is large.
9) except urine extracellular is also suitable for hydrothorax, ascites, sputum, cerebrospinal fluid and vaginal exfoliated inspection;
10) after patient's original urine cell dyes sheet mounting, can preserve for a long time;
11) patient that saves as for a long time that the foundation of Urinary cell detection data bank and original urine cell dye sheet follows up a case by regular visits to guarantee is provided.
12) feature of automatic thin sectioning cells is that disposable cell rejection tablet pipe design, high density Ficoll dilution prevent cell deposition and skewness, and stand apparatus can be used for a long time.Owing to there being rejection tablet liquid case, avoided the pollution in hydro-extractor.
Accompanying drawing explanation
Fig. 1, urinary cell micro staining analysis system schematic.
Fig. 2, urine sample cell separation apparatus,
Wherein, 1, freshly voided urine; 2, filter screen irrigator; 3, filter screen (30～300 order); 4, cross the urine cell of netting; 5, the strong point.
Fig. 3, urine cell gradient density centrifugation sampling apparatus,
Wherein, 1, urine cellular collector; 2, cross the arena after net; 3, urine cellular layer; 4, the Ficoll liquid of variable concentrations; 5, switch; 6, rubber stopper; 7, air metal catheter; 8, automatic control flow; 9, collect urine cellular layer; A, before urine cell collection device is centrifugal, B, controls plug, and C, after urine cell collection device density centrifugation.
The automatic pelletizer of Fig. 4,
Wherein, 1, base; 2, support; 3, acellular high density Ficoll waste liquid; 4, disposable plastic cell rejection tablet device; 5, rejection tablet device stationary installation; 6, circular rubber band; 7, microslide spiral fixator; 8, microslide; 9, exhausted bath box; 10, switch; 11, high density Ficoll urinates cell mixture;
12, waste liquid flows out pipeline; A, before rejection tablet, B, after rejection tablet.
The automatic pelletizer pelletizer of Fig. 5 outboard profile,
Wherein, 1, base; 2, support; 3, acellular high density Ficoll waste liquid; 4, disposable plastic cell rejection tablet device; 5, rejection tablet device stationary installation; 7, microslide spiral fixator; 9, exhausted bath box;
12, waste liquid flows out pipeline; 13 microslide cell holes.
Fig. 6 urinates cell automatic staining device sectional view,
Wherein, 1. admission gear, 2. air groove 3. stain groove 4. microslide rotating disks, 5. control panel 6. motors, 7. water mortise, 8. micro-stain pipettor, 9. air exhausting device 10. filtration units, 11. woven hoses, 12. Rinsing Areas.
Fig. 7 dyeing apparatus rotating disk figure, wherein, 1. microslide rotating disk, 2. microslide.
The quantitative test of Fig. 8 standard scale,
Wherein, 1, cell detection district, 2, cell detection scale, 3, label area.
Fig. 9 urinary cell micro dyeing flow.
Embodiment
Freshly voided urine 10～50ml, through the centrifugal 10min of 2000rpm, sediment mixes with 1ml Hanks liquid, cross 30～300 object filter screen (Fig. 2,3), under apocarpy, slowly inject 1ml arena Hanks liquid and in Ficoll, be made into the gradient of continuous density test tube of 1.095～1.055g/ml, the cell mass of sucking-off layering, through the centrifugal 10min of 2000rpm, resuspended with 0.5ml Hanks liquid.Adopt Hanks liquid to carry out after sample dilution, get 200 μ l urine cell samples and add 3～8ml Ficoll (1.065g/ml) solution to mix in the clean cell rejection tablet device of rear immigration, through the centrifugal 5min of 3000rpm, automatically make thin cell sheet (Fig. 4,5).Fixing dry rear standby with absolute ethyl alcohol.During dyeing, 1 microslide is placed on to automatic staining device [Fig. 6,7] in staining trough, input respectively each dyeing procedure, as Papanicolaou's vaginal smear technique, automatic staining device is controlled addition and the kind of dyeing liquor by the chip program of device, complete: 1) urine cell smear adds distilled water 100 μ l 2min, and distilled water flushing dries up; 2) haematine dye liquor 50 μ l15min, rinse with flowing water; 3) add 0.25% watery hydrochloric acid, 50 μ l to remove unnecessary haematine dye liquor in endochylemas, after 5 seconds, with flowing water, rinse; 4) add rare lithium carbonate solution 50 μ l alkalization 1min, flowing water rinses; 5) add 95% alcohol 100 μ l dehydration 2min, flowing water rinses; 6) add EA liquid 50 μ l dyeing 3min; 7) add 100 μ l95% alcohol washing 3 times, remove too much dye liquor; 8) add 100 μ l anhydrous alcohol dehydration 3min; 9) add 100 μ l dimethylbenzene 1min to dye, mounting.In above-mentioned dyeing course,
(1) natural language of HE dyeing course is:
1s haematine dye liquor; Wait for 210s; 210s tap water; 1s 0.5% hydrochloride alcohol; Wait for 7s; 210s tap water; 1s 0.5% ammoniacal liquor oil blackeite; Wait for 45s; 210s tap water; 210s distilled water; 1s 0.5% Yi Hongshui; Wait for 180s; 15s distilled water; 23s 80% alcohol; 23s 95% alcohol; 90s 100% anhydrous alcohol; 1s dimethylbenzene; Wait for 90s; 1s dimethylbenzene; Wait for 240s; 2s neutral gum.
(2) to translate above-mentioned natural language be following sequence to control module,
F2_L2_S1.##__210^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F10_L4_S1.##__7^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F11_L4_S1.##__45^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F30_L20_L21_S210.F12_L5_S1.##__180^0.F30_L20_L21_S15.F13_L9_L10_L11_S23.F5_L9_L10_L11_S23.F7_L9_L10_L11_S90.F8_L3_S1.##__90^30.F8_L3_S1.##__240^60.F9_L4_S2
Event sheet is separated by point (.); The event sheet starting with alphabetical F, represents to need that system respond with stain or rinse and move; The event sheet that the character # of take starts is waiting event, does not need system to provide action.As: F2_L2_S1.##__210^0.F5_L9_L10_L11_S23
Wherein, haematine dye liquor is defined as No. 2 functions (F2) and is assemblied on No. 2 positions (L2) this event effect 1 second (S1); Waiting event starts (##__) with two No. # and two underscores and adds the stand-by period, and ' ^ ' number represents that heel numeral waits for error of fragment permission for this, so ##__210^0 represents to wait for 210 seconds, and error should be controlled at ± in 0 second; With 80% alcohol, rinse 23 seconds, be expressed as F5_L9_L10_L11_S23, wherein 80% alcohol functionality number 5 (F5), are arranged on 9,10, No. 11 positions (_ L9_L10_L11), act on 23 seconds (S23).
The output sequence form of control module is as follows:
Begin ###
C：0 BP：1
S TF0 L2 Fn2
R TF1 L2 Fn2
C：1 BP：0
S TF3 L2 Fn2
R TF4 L2 Fn2
C：9 BP：14
S TF14 L20 Fn30
R TF19 L20 Fn30
C：6 BP：11
S TF213 L12 Fn29
S TF216 L13 Fn29
R TF423 L12 Fn29
R TF426 L13 Fn29
End ###
In output sequence, concrete response action of every line display.Specifically be divided into three kinds: C character starts, represent master rotation.S character starts, and presentation function is opened.R character starts, and presentation function is closed.In " C:9 BP:14 ", 9 expression systems are the reserved rotational time of this rotation process.14 represent take that No. 0 Ran Pian position is as with reference to master is turned to position 14.Sequence is as the input of system dyeing action processing module, and is finally converted into mechanical action;
After purifying, the treated device of dyeing liquor and waste gas discharges; Dye sheet and insert high-resolution color numeral micrometron platform, under computer keyboard or mouse control, by microscope, video high Definition CCD image acquisition device, transmit image to LCD colorful display screen, mouse is selected staining cell region, mark kind, is taken pictures; Kind and the number of computing machine Auto-counting urine cell, analysis result is printed and is stored.Analysis result shows: urine cell dyeing is clear, is evenly distributed, and is easy to identification and quantitative.
Freshly voided urine 10～50ml, through the centrifugal 10min of 2000rpm, sediment mixes with 1ml Hanks liquid, cross 30～300 object filter screen (Fig. 2,3), under apocarpy, slowly inject 1ml arena Hanks liquid and in Ficoll, be made into the gradient of continuous density test tube of 1.095～1.055g/ml, the cell mass of sucking-off layering, through the centrifugal 10min of 2000rpm, resuspended with 0.5ml Hanks liquid.Adopt Hanks liquid to carry out after sample dilution, get 200 μ l urine cell samples and add 3～8ml Ficoll (1.065g/ml) solution to mix in the clean cell rejection tablet device of rear immigration, through the centrifugal 5min of 3000rpm, automatically make thin cell sheet (Fig. 4,5).Fixing dry rear standby with absolute ethyl alcohol.During dyeing, 24 microslides are placed on to automatic staining device [Fig. 6,7] in staining trough, input respectively each dyeing procedure, as Papanicolaou's vaginal smear technique, automatic staining device is controlled addition and the kind of dyeing liquor by the chip program of device, complete: 1) urine cell smear adds distilled water 100 μ l 2min, and distilled water flushing dries up; 2) haematine dye liquor 50 μ l15min, rinse with flowing water; 3) add 0.25% watery hydrochloric acid, 50 μ l to remove unnecessary haematine dye liquor in endochylemas, after 5 seconds, with flowing water, rinse; 4) add rare lithium carbonate solution 50 μ l alkalization 1min, flowing water rinses; 5) add 95% alcohol 100 μ l dehydration 2min, flowing water rinses; 6) add EA liquid 50 μ l dyeing 3min; 7) add 100 μ l95% alcohol washing 3 times, remove too much dye liquor; 8) add 100 μ l anhydrous alcohol dehydration 3min; 9) add 100 μ l dimethylbenzene 1min to dye, mounting.In above-mentioned dyeing course,
It is following sequence that control module is translated above-mentioned multi-disc natural language,
In clobal table descriptive system, the needed dyeing course of institute's sheet of having illicit sexual relations of current loading, is comprised of multirow, and line number is the maximum load amount of system.
0！；
1：F2_L2_S1.##__210^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F10_L4_S1.##__7^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F11_L4_S1.##__45^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F30_L20_L21_S210.F12_L5_S1.##__180^0.F30_L20_L21_S15.F13_L9_L10_L11_S23.F5_L9_L10_L11_S23.F7_L9_L10_L11_S90.F8_L3_S1.##__90^30.F8_L3_S1.##__240^60.F9_L4_S2；
2：F2_L2_S1.##__210^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F10_L4_S1.##__7^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F11_L4_S1.##__45^0.F29_L12_L13_L14_L15_L16_L17_L18_L19_S210.F30_L20_L21_S210.F12_L5_S1.##__180^0.F30_L20_L21_S15.F13_L9_L10_L11_S23.F5_L9_L10_L11_S23.F7_L9_L10_L11_S90.F8_L3_S1.##__90^30.F8_L3_S1.##__240^60.F9_L4_S2；
3：F30_L20_L21_S5.F14_L5_S1.##__1200^0.F29_L12_L13_L14 L15_L16_L17_L18_L19_S60.F15_L5_S1.##__15^5.F29_L12_L13_L14_L15_L16_L17_L18_L19_S90.F30_L20_L21_S7.F16_L6_S1.##__240^60.F29_L12_L13_L14_L15_L16_L17_L18_L19_S90.F30_L20_L21_S7.F17_L6_S1.##__60^0.F18_L6_S7.F9_L4_S2；
4！；
5！；
6！；
7！；
8！；
Upper table has been described situation when system loading multi-disc is dyed sheet.Line number opens from zero, and every row is started by line number, heel dyeing course descriptor, and with branch (; ) ending.If certain row unloaded is dyed sheet, by exclamation mark (! ) replacement dyeing course descriptor.In upper table, first trip " 0! ; " representing that No. 0 position of rotating disk do not dye sheet, the second No. 1, line display position has been loaded and has been dyed sheet, lists file names with the dyeing course that this dyes sheet requirement, and same, 2,6 positions have also been loaded and have been dyed sheet.Here only for describing the problem, unlisted whole length of a game shows.In fact, clobal table can be comprised of the line number of the maximum load amount of system (24).Clobal table is the input of control module algorithm.
The output sequence form of control module is as follows:
Begin ###
C：0 BP：1
S TF0 L2 Fn2
R TF1 L2 Fn2
C：1 BP：0
S TF3 L2 Fn2
R TF4 L2 Fn2
C：9 BP：14
S TF14 L20 Fn30
R TF19 L20 Fn30
C：6 BP：11
S TF213 L12 Fn29
S TF216 L13 Fn29
R TF423 L12 Fn29
R TF426 L13 Fn29
End ###
In output sequence, concrete response action of every line display.Specifically be divided into three kinds: C character starts, represent master rotation.S character starts, and presentation function is opened.R character starts, and presentation function is closed.In " C:9 BP:14 ", 9 expression systems are the reserved rotational time of this rotation process.14 represent take that No. 0 Ran Pian position is as with reference to master is turned to position 14.Sequence is as the input of system dyeing action processing module, and is finally converted into mechanical action;
After purifying, the treated device of dyeing liquor and waste gas discharges, dye sheet and insert high-resolution color numeral micrometron platform, under computer keyboard or mouse control, by microscope, video high Definition CCD image acquisition device, transmit image to LCD colorful display screen, mouse is selected staining cell region, mark kind, is taken pictures, kind and the number of computing machine Auto-counting urine cell, analysis result is printed and is stored.
Claims (8)
1. a urinary cell micro staining analysis device, is characterized in that described analytical equipment comprises urine sample cell separation apparatus, urine cell density gradient centrifugation sampling apparatus, automatic pelletizer, automatic staining device and dyeing liquor treating apparatus, high Definition CCD image acquisition device, high-resolution color numeral micrometron platform and microscope, computer control system and controls software, LCD colorful display screen and color printer; Wherein, described urine cell density gradient centrifugation sampling apparatus forms by urinating cellular collector, switch, rubber stopper, air metal catheter, automatic control flow and control plug, and the Ficoll liquid of variable concentrations is housed in described urine cell density gradient centrifugation sampling apparatus; Described automatic pelletizer is comprised of base, support, disposable plastic cell rejection tablet device, rejection tablet device stationary installation, circular rubber band, microslide spiral fixator, microslide, exhausted bath box, switch, high density Ficoll urine cell mixture, waste liquid outflow pipeline and microslide cell hole; Described automatic staining device is comprised of admission gear, air groove, dye liquor tank, microslide rotating disk, control panel, motor, Shui Cao, micro-stain pipettor, air exhausting device, filtration unit, woven hose and Rinsing Area; Described automatic staining device dyes sheet under the software control of slide Dyeing control; Described slide Dyeing control software consists of length of a game's axle, and introduces the probabilistic search module based on genetic algorithm; Described length of a game axle adopts take the list structure that single dyeing action is minimum treat node, in logic mark each time etching system dyeing behavior, and by one group of function, provided the interface of insertion, query manipulation; On the basis of described genetic algorithm Shi length of a game axle, the appropriate dispatching sequence of structure.
2. urinary cell micro staining analysis device claimed in claim 1, is characterized in that described urine sample cell separation apparatus consists of filter screen irrigator, filter screen and screen support point; Filter screen is wherein 30～300 orders.
3. the urinary cell micro staining analysis device described in claim 1 or 2, is characterized in that the urinary cell micro staining analysis carrying out with described urinary cell micro staining analysis device comprises the steps:
1) after urine sample separation to be measured, urinate cell sample,
Urine sample to be measured enters urine cell gradient density centrifugation sampling apparatus, after the automatic separation of urine sample cell separation apparatus, proceeds to urine cell density gradient centrifugation sampling apparatus, through density gradient centrifugation collection urine cell liquid;
2) urine cell automatic thin layer film-making,
The urine cell liquid gathering, after dilution, moves to rejection tablet film-making automatically in automatic pelletizer;
3) urine cell sheet automatic fine dyeing,
The urine cell sheet making moves into automatic staining device and dye sheet under the software control of slide Dyeing control;
4) gather urine cell and dye picture
Urine cell dyes sheet and inserts high-resolution color numeral micrometron platform, through micro-platform control software control automatically, through microscope, video high Definition CCD image acquisition device, is sent to the image on LCD colorful display screen,
5) urine cell classification mark and image photographic, qualitative and quantitative analysis,
The urine cell of step 4) dyes picture and urinate cell classification mark and image photographic behind the urine cell image Real-time Collection automatic analysis selected staining cell of quantitative Treatment software region, automatically carries out qualitative and quantitative analysis;
6) print urine test result picture and text report, grouping of data management.
4. urinary cell micro staining analysis device according to claim 3, is characterized in that, described step 1) adopts the Ficoll density gradient solution of 1.025～1.095g/ml through density gradient centrifugation collection urine cell liquid be urine cell collection liquid.
5. urinary cell micro staining analysis device according to claim 3, is characterized in that described step 2) automatic rejection tablet film-making in adopt the Ficoll solution of 1.010～1.050g/ml suspend to extract urine cellular layer.
6. urinary cell micro staining analysis device according to claim 3, is characterized in that, the dye liquor of the slide dyeing of described step 3) is the dyeing applied sample amount of 10～300 μ l.
7. urinary cell micro staining analysis device according to claim 3, it is characterized in that circular stage and microslide in the automatic staining device described in step 3) are coupled, circular stage is placed 24～364 microslides simultaneously, adopt rotation mode to carry out application of sample design, the kind of simultaneously controlling dyeing application of sample is 24～64 kinds, the application of sample time is 0.1～1 second, and dyeing time is 2 seconds～48 hours.
8. urinary cell micro staining analysis device according to claim 3, the quantitative test process that it is characterized in that described step 5) is: urine cell sample extracts cell device, thin layer rejection tablet device, dyeing circular area by filter screen tripping device, gradient density centrifugal, and to have scale to proofread and correct, determine that computing machine after target cell completes the counting of target cell automatically quantitative, and described dyeing circular area diameter is 0.25～1.0mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810040597.7A CN101629946B (en) | 2008-07-15 | 2008-07-15 | Urinary cell micro staining analysis method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810040597.7A CN101629946B (en) | 2008-07-15 | 2008-07-15 | Urinary cell micro staining analysis method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101629946A CN101629946A (en) | 2010-01-20 |
| CN101629946B true CN101629946B (en) | 2014-04-09 |
Family
ID=41575127
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200810040597.7A Active CN101629946B (en) | 2008-07-15 | 2008-07-15 | Urinary cell micro staining analysis method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101629946B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103149070B (en) * | 2013-03-11 | 2014-10-08 | 黄石市第一医院 | Automatic slide stainer |
| CN104359746B (en) * | 2014-11-19 | 2018-03-16 | 白银市第一人民医院 | Cell mass collector and the method for collecting cell in humoral specimen |
| CN111175099B (en) * | 2019-12-09 | 2024-06-25 | 湖北泰康医疗设备有限公司 | Extraction and tabletting method for body fluid cells for detecting bladder cancer |
| CN111180058B (en) * | 2020-01-07 | 2023-05-12 | 同济大学 | Automatic urinary cell staining efficiency optimization method |
| CN112525633B (en) * | 2020-11-20 | 2022-12-20 | 深路医学科技(武汉)有限公司 | Tablet making control method and tablet making machine |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1319643A (en) * | 1969-06-26 | 1973-06-06 | Information Utilization Corp | Preparing cellular specimens for subsequent testing for the pre sence of diseased cells and method and apparatus for carrying out the subsequent testing |
| US6780380B2 (en) * | 2001-01-16 | 2004-08-24 | Triangle Biomedical Sciences, Inc. | Tissue processor |
| CN1727902A (en) * | 2003-09-04 | 2006-02-01 | 合肥新月电子技术有限责任公司 | Full-automatic urine analysis system |
-
2008
- 2008-07-15 CN CN200810040597.7A patent/CN101629946B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1319643A (en) * | 1969-06-26 | 1973-06-06 | Information Utilization Corp | Preparing cellular specimens for subsequent testing for the pre sence of diseased cells and method and apparatus for carrying out the subsequent testing |
| US6780380B2 (en) * | 2001-01-16 | 2004-08-24 | Triangle Biomedical Sciences, Inc. | Tissue processor |
| CN1727902A (en) * | 2003-09-04 | 2006-02-01 | 合肥新月电子技术有限责任公司 | Full-automatic urine analysis system |
Non-Patent Citations (4)
| Title |
|---|
| 刘俊彬等.流行性出血热患者尿液沉渣超活体染色镜检及其临床意义.《第四军医大学学报》.1992,第13卷(第1期),全文. |
| 尿细胞成分染色检查对小儿肾脏疾病诊断的意义;项彤芳等;《实用儿科杂志》;19871231;第2卷(第2期);全文 * |
| 流行性出血热患者尿液沉渣超活体染色镜检及其临床意义;刘俊彬等;《第四军医大学学报》;19920131;第13卷(第1期);全文 * |
| 项彤芳等.尿细胞成分染色检查对小儿肾脏疾病诊断的意义.《实用儿科杂志》.1987,第2卷(第2期),全文. |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101629946A (en) | 2010-01-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101629946B (en) | Urinary cell micro staining analysis method | |
| Chien et al. | Urine sediment examination: a comparison of automated urinalysis systems and manual microscopy | |
| CN101560544B (en) | Method for integrative detection of cells | |
| CN102393705B (en) | Sample formed component analyzer automatic detection control apparatus and control method thereof | |
| CN115266540A (en) | Optical measurement performed on a sample | |
| CN109612911B (en) | Full-automatic sperm cell detector | |
| RU2007137654A (en) | METHOD, DEVICE AND SYSTEM FOR VOLUMETRIC CALCULATION OF WHITE BLOOD CELLS | |
| CN202770842U (en) | Urine analysis device | |
| CN101587109B (en) | Determination method and intruments of hexavalent chromium | |
| CN101560543B (en) | Cell staining reagent and preparation method and application thereof in cell staining | |
| CN101963612A (en) | Detection kit for sperms and check method thereof | |
| CN201060190Y (en) | Full-automatic dejecta analytical equipment | |
| CN103852368A (en) | Mitochondria DNA observation through MTG-DAPI double-staining of semi-thin sections of cucumber pollen | |
| Toma et al. | Comparison of 4 fixation and staining methods for the cytologic evaluation of ear canals with clinical evidence of ceruminous otitis externa | |
| Dolled-Filhart et al. | Automated analysis of tissue microarrays | |
| US20030138877A1 (en) | Automated staining and decolorization of biological material | |
| CN116448988A (en) | Industrial park soil pollution monitoring system and method | |
| CN202257125U (en) | Automatic detection controller for sample visible component analytical instrument | |
| Gandjeva et al. | Lung histological methods | |
| CN102839208B (en) | Absolute quantification method for fluorescence in-situ hybrid cell counting | |
| CN112697779A (en) | Water quality online monitoring system and method and water sample storing and taking device and method thereof | |
| CN201305591Y (en) | Microorganism intelligent identification equipment | |
| CN201266164Y (en) | Device for counting urine cell tittle dyeing | |
| CN216095204U (en) | A belt cleaning device and push jack dyeing machine for push jack dyeing machine | |
| CN218885746U (en) | Water quality testing device that short-term test |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |