Novel chromosome analysis smear processing system and method

Abstract

The invention relates to the field of biomedical detection, and provides a novel chromosome analysis smear processing system and method. The system is based on a full-automatic sample treatment platform and comprises an automatic cell inoculation and culture module, an automatic cell harvesting module, an automatic cell dropping module and a cell genetic information management system, the single-batch treatment flux is not less than 192, and the system can be expanded. And precise control of each link is realized through technologies of a high-precision split charging pump, electric field assisted cell separation and the like. The method comprises the steps of automatic inoculation, constant-temperature culture, electric field assisted harvesting, precise dropping and data monitoring and analysis. Compared with a traditional mode, the method solves the problems of complicated equipment, low flux, insufficient standardization and the like, has the advantages of high processing flux, high standardization and informatization degree, small operation error, intelligent quality control and the like, and can effectively improve the cytogenetics analysis efficiency and accuracy.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical detection, and in particular to a novel chromosome analysis smear processing system and method. Background Art

[0002] Cytogenetic analysis plays a vital role in prenatal diagnosis, occupational disease examination, reproductive medicine, blood disease diagnosis and third-party laboratory testing. The classic cytogenetic analysis sample processing process includes sample inoculation, cell culture, cell harvesting, preparation and staining. However, the traditional processing method has many disadvantages:

[0003] 1. Complex equipment and low throughput: Each processing link relies on different equipment to complete. The large number of equipment not only takes up a lot of space, but also leads to cumbersome operation procedures. In addition, the overall processing throughput is low, which is difficult to meet the growing testing needs. For example, the traditional method can only process a limited number of samples in a single batch. When faced with large-scale testing tasks, the efficiency is extremely low, which seriously affects the progress of testing.

[0004] 2. Insufficient standardization and informatization: Due to the involvement of multiple devices and manual operations, it is difficult to unify the processing standards between different operators and different laboratories, resulting in poor repeatability and comparability of experimental results. At the same time, information from each link cannot be effectively integrated and shared, making it difficult to achieve informatization management of the entire process, which is not conducive to data analysis and traceability.

[0005] 3. Complex operation and large errors: There are many manual operation steps, and the technical requirements for operators are high, which is easy to introduce human errors. Taking manual cell harvesting as an example, slight deviations in factors such as culture time, hypotonic treatment, and colchicine action time will have a significant impact on cell quality. If the culture time is short or the subject is older, the division phase will be reduced; the hypotonic fluid is not sufficiently dispersed, and the cells are clumped; bacterial contamination will lead to an increase in the number of bacteria; the long action time of colchicine will increase the proportion of chromosomes that are too short; long hypotonic time will cause the chromosomes to disperse excessively or even break. These factors will affect the accuracy of the final test results.

[0006] 4. Technical limitations in some key links: In the automatic cell inoculation and culture module, there is a lack of clear standards and effective means for the precise control of culture medium packaging accuracy, inoculation volume accuracy, and cell culture conditions, making it difficult to ensure the consistency and accuracy of the experiment. In the automatic cell harvesting module, the traditional harvesting method has uncertain influencing factors that cannot be effectively eliminated, resulting in low sample processing throughput, large processing differences between samples, and a lack of quantitative effect comparison data. The continuous dripping method of the automatic dripping module lacks a detailed description of the mechanical structure and principles, and the temperature and humidity control parameters are unclear, making it difficult to achieve efficient and stable film production. The technical details of the docking of the cell genetic information management system with the external system and the internal algorithm logic are not perfect, and cannot meet the needs of full-process information management and quality control.

[0007] To solve the above problems, the present invention provides a novel chromosome analysis smear processing system and method, which aims to integrate cell inoculation, culture, harvesting, smear dripping and information management into a single device through a fully automatic integrated design, improve the processing throughput and standardization level, ensure the accuracy and reliability of the test results, and meet the needs of various related fields for efficient and precise detection of cytogenetic analysis. Summary of the invention

[0008] In order to solve the above technical problems, the present invention provides a novel chromosome analysis smear processing system and method, which integrates cell inoculation, culture, harvesting, smear dropping and information management into a single device through a fully automatic integrated design, thereby significantly improving the processing throughput and standardization level.

[0009] A novel chromosome analysis smear processing system of the present invention comprises: an automatic cell inoculation and culture module, an automatic cell harvesting module, an automatic drop film module, and a cell genetic information management system; the system is based on a fully automatic sample processing platform, has the ability to process a single batch of no less than 192 samples, and the throughput is expandable; the automatic cell inoculation and culture module is used for culture medium packaging, sample inoculation and cell culture; the automatic cell harvesting module integrates a variety of cell harvesting processes; the automatic drop film module is used for smear preparation; the cell genetic information management system is connected to an external system and monitors the whole process data, and the above-mentioned fully automatic sample processing system uses the SPS-HS-01 model processing system equipment of Shanghai Lechen Biotechnology Co., Ltd.

[0010] Preferably, the automatic cell inoculation and culture module is equipped with a high-precision filling pump and a constant temperature culture unit, the culture medium filling accuracy reaches ±0.15ml, the inoculation volume range is 0.3-0.5ml, the culture temperature is controlled at 37℃±0.5℃, and the carbon dioxide concentration in the culture environment is controlled at 5%.

[0011] Preferably, the high-precision filling pump achieves high-precision filling through volumetric quantitative filling technology, which utilizes a high-precision piston pump structure to accurately control the filling volume of the culture medium by controlling the displacement of the piston under the precise drive of the motor; the constant temperature culture unit utilizes a PID (proportional-integral-differential) temperature control circuit in combination with a heating wire and a cooling plate to maintain a constant temperature, and the temperature sensor monitors the culture environment temperature in real time and feeds back the signal to the PID temperature control circuit, which adjusts the working state of the heating wire and the cooling plate, thereby achieving precise control of the culture temperature.

[0012] Preferably, the automatic cell harvesting module integrates centrifugation, hypotonic treatment and fixation processes, and adopts two fixation processes, with 6 ml of fixative added each time and a fixation time of 20-30 minutes.

[0013] Preferably, the innovative harvesting method of the automatic cell harvesting module eliminates uncertainties through electric field-assisted cell separation technology, specifically: during the centrifugation process, an alternating electric field with a frequency of 20-50 Hz and an intensity of 0.5-2 V / cm is applied, so that the cells settle more orderly under the action of the electric field force, reducing cell aggregation and loss; the centrifugal speed is 1800 rpm, and the centrifugation time is 6 minutes; at the same time, during the hypotonic treatment and fixation process, the presence of the electric field promotes full contact between chemical reagents and cells, improves the consistency of the treatment effect, and thereby improves the sample processing throughput and reduces the processing differences between samples.

[0014] Preferably, the automatic film dropping module adopts a continuous film dropping mechanical structure and a temperature and humidity control device, the temperature and humidity control accuracy is ±0.25°C and ±2%RH, RH is relative humidity, and the film dropping height is 10-30cm.

[0015] Preferably, the dripper of the continuous drip sheet mechanical structure adopts a quartz capillary micro-dripper, which has good chemical stability and low surface energy, and can effectively reduce liquid residue and wall hanging phenomenon; the driving mechanism is piezoelectric ceramic drive, by applying a changing voltage to the piezoelectric ceramic, causing it to produce a slight deformation, thereby pushing the liquid out of the dripper; by optimizing the inner diameter, length and shape of the internal flow channel of the dripper, using Poiseuille's law in fluid mechanics, the flow rate and droplet size of the dripping liquid are accurately controlled to ensure the stability of the dripping liquid.

[0016] Preferably, the cell genetic information management system transmits data to the LIS / HIS system in real time, and the file format of the data transmission is set to the TXT file format, but is not limited to this format, and the algorithm based on threshold judgment and statistical analysis is used internally for data processing and quality control. The specific judgment rules are: the normal threshold range of cell survival rate is set to 70%-95%, the normal range of chromosome dispersion is 7-9 levels (using the industry's general dispersion rating standard), and the normal threshold range of chromosome aberration rate is 0-3%. When the cell survival rate is lower than 70% or higher than 95%, the chromosome dispersion is lower than 7 levels or higher than 9 levels, and the chromosome aberration rate is higher than 3%, the system automatically determines that the data is abnormal and triggers an early warning. The statistical method is: the cell survival rate, chromosome dispersion and aberration rate in each batch of experimental data are collected, and the mean value, standard deviation and coefficient of variation of each index are calculated. If the coefficient of variation of an indicator is greater than 10%, the stability of the experimental data of this batch is judged to be poor, and the system prompts the operator to check the experimental equipment or re-perform the experiment; if the average value of an indicator of consecutive specified batches exceeds the normal threshold range, the system will also issue an early warning and prompt relevant inspections and adjustments. The selection of the specified batches needs to consider many factors, including experimental stability and reliability, risk tolerance, and historical data and experience.

[0017] A novel chromosome analysis smear processing method comprises the following steps:

[0018] Step 1: Use the high-precision dispensing pump in the automatic cell inoculation and culture module to automatically dispense the culture medium, and use the micro-injection pump combined with a high-precision flow sensor to control the inoculation volume within the range of 0.3-0.5 ml for sample inoculation;

[0019] Step 2: Place the inoculated sample in the constant temperature culture unit of the automatic cell inoculation and culture module, use the PID temperature control circuit combined with the heating wire and the cooling sheet to maintain the culture temperature at 37°C ± 0.5°C, and use the gas concentration control device to maintain the carbon dioxide concentration in the culture environment at 5% for constant temperature culture;

[0020] Step 3: Through the automatic cell harvesting module, electric field-assisted cell separation technology is used to first perform centrifugation operation, and centrifuge at a speed of 1800 rpm for 6 minutes under an alternating electric field environment with a frequency of 20-50 Hz and an intensity of 0.5-2 V / cm to allow the cells to initially settle; then 0.075 mol / L potassium chloride is added as a hypotonic solution for hypotonic treatment, and the treatment time is 10 minutes; finally, two fixation processes are performed, and each time the amount of fixative (methanol: glacial acetic acid = 3:1 mixed solution) added is 6 ml, and the fixation time is 20-30 minutes to complete the cell harvest;

[0021] Step 4: Use the continuous film drop mechanical structure and temperature and humidity control device of the automatic film drop module to make films. The piezoelectric ceramic drives the quartz capillary micro-droplet head to achieve continuous film drop. The film drop flow rate is controlled to 0.2ml / s, and the droplet diameter is controlled to 1.0mm±0.05mm. At the same time, the temperature and humidity are controlled at 20-40℃ and 25%-75%RH respectively by using the temperature and humidity control device combined with the compressor and the semiconductor. The height of the film drop is 10-30cm;

[0022] Step 5: Through the cell genetic information management system, using an algorithm based on threshold judgment and statistical analysis, upload the data during the experiment in real time, and monitor quality control parameters such as cell survival rate, chromosome dispersion and aberration rate. When the data is abnormal, timely warnings will be issued and the corresponding processing flow will be initiated.

[0023] Preferably, in step five, if the cell survival rate is lower than 70%, the system prompts to check whether the culture conditions are abnormal; if the chromosome dispersion is lower than level 7, it prompts to optimize the droplet operation parameters; if the chromosome aberration rate is higher than 3%, it prompts to check whether the experimental sample is contaminated or whether the experimental process is wrong.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] 1. Improved throughput and integration: Fully automatic integrated design, single batch processing throughput is not less than 192 and can be expanded, integrating multiple links into a single device to solve the problems of complex equipment and low throughput.

[0026] 2. Standardization and information enhancement: unify processing standards to improve the repeatability and comparability of results; realize information management of the entire process to facilitate data traceability and analysis.

[0027] 3. Operation and precision optimization: reduce manual operations and errors; each module is precisely controlled, such as the inoculation and culture module accurately controls the packaging and culture conditions, the harvesting module uses new technologies to improve the processing effect, and the drop film module stabilizes the film production.

[0028] 4. Intelligent quality control: The genetic information management system monitors key parameters in real time, issues abnormal warnings and prompts processing to ensure accurate and reliable results. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 is a schematic diagram of the system architecture of the present invention;

[0030] Figure 2 is a flow chart for implementing the method of the present invention; DETAILED DESCRIPTION

[0031] The specific implementation of the present invention is further described in detail below in conjunction with the accompanying drawings and examples. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0032] like Figure 1 to Figure 2 As shown,

[0033] 1. Embodiment 1

[0034] 1. Experimental preparation

[0035] This example uses the SPS-HS-01 fully automatic sample processing system of Shanghai Lechen Biotechnology Co., Ltd. to carry out experiments, aiming to process 192 samples for chromosome analysis smear preparation. In the preparation stage, all modules of the system are fully checked and calibrated to ensure that the equipment is in the best operating state.

[0036] In the automatic cell inoculation and culture module, the high-precision filling pump uses volumetric quantitative filling technology to drive a high-precision piston pump with a motor to accurately control the piston displacement to achieve the filling of the culture medium. The 10 filling processes were recorded, and the results showed that the filling accuracy was stable at ±0.15ml. The specific data is shown in Table 1 below:

[0037] Table 1:

[0038]

[0039]

[0040] Through the combination of a micro-injection pump and a high-precision flow sensor, the inoculation volume was precisely controlled at 0.3 ml, and the inoculation operation of 192 samples was completed, with the inoculation volume error controlled within a very small range.

[0041] (II) Cell culture

[0042] The inoculated samples were quickly placed in a constant temperature culture unit. The PID temperature control circuit was combined with the heating wire and the cooling sheet for temperature control, and the temperature sensor monitored the culture environment temperature in real time. During the 24-hour continuous culture process, the temperature data was recorded every hour, and the culture temperature was stably maintained at around 37°C, with a fluctuation range of ±0.5°C, which fully met the experimental requirements. At the same time, the carbon dioxide concentration was accurately controlled at 5% through the gas concentration control device to create a stable environment for cell growth.

[0043] (III) Cell harvesting

[0044] The automatic cell harvesting module was started, and electric field-assisted cell separation technology was used. During the centrifugation process, an alternating electric field with a frequency of 30 Hz and an intensity of 1 V / cm was applied, and the centrifuge was centrifuged at a speed of 1800 rpm for 6 minutes. After the centrifugation, 0.075 mol / L potassium chloride solution was added for 10 minutes of hypotonic treatment. Then two fixation processes were performed, each time 6 ml of fixative solution (methanol: glacial acetic acid = 3:1 mixture) was added, and each fixation time was 25 minutes.

[0045] To verify the effect of this harvesting method, 10 samples were randomly selected and compared with samples processed by the traditional harvesting method to detect cell purity, integrity and harvest volume. The results are shown in Table 2 below:

[0046] Table 2:

[0047]

[0048]

[0049] It can be clearly seen from the data comparison that the harvesting method of the present invention is superior to the traditional method in terms of cell purity, integrity and harvest volume.

[0050] (IV) Automatic film drop

[0051] The automatic film drop module starts working, and the quartz capillary micro-droplet head of the continuous film drop mechanism is driven by piezoelectric ceramics to drop film. By optimizing the dropper parameters, the drip state is accurately controlled according to Poiseuille's law. The drop flow rate is set to 0.2ml / s. During the continuous drop process of 100 times, the droplet diameter is measured every 10 times. The results are shown in Table 3 below:

[0052] Table 3:

[0053]

[0054]

[0055] It can be seen that the droplet diameter is stably controlled at about 1.0 mm with minimal deviation. The temperature and humidity control device controls the ambient temperature at 25°C, the relative humidity at 50%, and the droplet height at 20 cm to ensure an efficient and smooth film making process.

[0056] (V) Data monitoring and analysis

[0057] The cell genetic information management system monitors the entire experimental process in real time and transmits the experimental data to the LIS / HIS system in real time in TXT file format. The system processes and controls the data quality based on threshold judgment and statistical analysis algorithms. In this experiment, cell survival rate, chromosome dispersion and aberration rate were monitored in real time.

[0058] After the experiment, the data of 192 samples were statistically analyzed to calculate the mean, standard deviation and coefficient of variation of each indicator. The results showed that the mean cell survival rate was 86%, the standard deviation was 2.5%, and the coefficient of variation was 2.91%; the mean chromosome dispersion was 8.2, the standard deviation was 0.4, and the coefficient of variation was 4.88%; the mean chromosome aberration rate was 2.8%, the standard deviation was 0.8%, and the coefficient of variation was 28.57%. All indicators were within the normal threshold range, and the experimental data were stable and reliable.

[0059] 2. Embodiment 2

[0060] 1. Experimental preparation

[0061] 250 samples were processed using the SPS-HS-01 fully automatic sample processing system. In the automatic cell inoculation and culture module, a high-precision dispensing pump dispensed the culture medium, and the dispensing accuracy was stabilized at ±0.15ml after multiple tests. The inoculation volume was set to 0.4ml, and all sample inoculations were completed with precise control through relevant equipment.

[0062] (II) Cell culture

[0063] The inoculated samples were placed in a constant temperature culture unit, and the culture temperature was set at 36.8°C. During the culture process, the temperature data was recorded every 30 minutes, and the temperature fluctuation was always controlled within the range of ±0.5°C, and the carbon dioxide concentration was stably maintained at 5%.

[0064] (III) Cell harvesting

[0065] The automatic cell harvesting module was running. In the electric field-assisted cell separation technology, the electric field frequency was set to 40 Hz, the intensity was 1.5 V / cm, the centrifuge speed was 1800 rpm, and the centrifugation was performed for 6 minutes. The hypotonic treatment used a 0.075 mol / L potassium chloride solution for 10 minutes. In the fixation process, the amount of fixative added each time was 6 ml, and the fixation time was 22 minutes and 28 minutes, respectively.

[0066] 15 samples were randomly selected to detect cell-related indicators and compared with Example 1. The data are shown in Table 4 below:

[0067] Table 4:

[0068]

[0069]

[0070] Compared with Example 1, the differences in various indicators are small, indicating that the method of the present invention has good stability.

[0071] (IV) Automatic film drop

[0072] When the automatic film drop module is working, the film drop flow rate is set to 0.2ml / s, and the droplet diameter is controlled within the range of 1.0mm±0.05mm. The temperature and humidity control device controls the temperature at 30℃, the relative humidity at 40%, and the droplet height is adjusted to 25cm. The flow rate and droplet diameter during the continuous film drop process are monitored, and the data show that the flow rate is stable at 0.2ml / s, and the droplet diameter fluctuation is within the allowable range.

[0073] (V) Data monitoring and analysis

[0074] The cell genetic information management system transmits the experimental data to the LIS / HIS system in real time in TXT file format. The experimental data were statistically analyzed, and the average cell survival rate was 87%, the standard deviation was 2%, and the coefficient of variation was 2.3%; the average chromosome dispersion was 8.3 levels, the standard deviation was 0.3 levels, and the coefficient of variation was 3.61%; the average chromosome aberration rate was 2.6%, the standard deviation was 0.7%, and the coefficient of variation was 26.92%. The experimental data is stable and meets the quality control requirements.

[0075] 3. Embodiment 3

[0076] 1. Experimental preparation

[0077] The SPS-HS-01 fully automatic sample processing system was selected to process 300 samples. The high-precision dispensing pump of the automatic cell inoculation and culture module accurately dispensed the culture medium, with a dispensing accuracy of ±0.15ml. The inoculation volume was set to 0.5ml to complete the inoculation of all samples.

[0078] (II) Cell culture

[0079] The inoculated samples were placed in a constant temperature culture unit and the culture temperature was maintained at 37.2°C. During the culture period, the temperature data was recorded every 2 hours, the temperature fluctuation was within ±0.5°C, and the carbon dioxide concentration was stabilized at 5%.

[0080] (III) Cell harvesting

[0081] The automatic cell harvesting module was running, the electric field frequency of the electric field-assisted cell separation technology was 25 Hz, the intensity was 0.8 V / cm, and the centrifuge was centrifuged at 1800 rpm for 6 minutes. The hypotonic treatment used 0.075 mol / L potassium chloride solution for 10 minutes. In the fixation process, 6 ml of fixative solution was added each time, and the fixation time was 30 minutes.

[0082] 20 samples were randomly selected to test cell indicators, and the data are shown in Table 5 below:

[0083] Table 5:

[0084]

[0085]

[0086] The cell harvesting effect was good, and the variability between samples was small.

[0087] (IV) Automatic film drop

[0088] The automatic film drop module is started, the film drop flow rate is maintained at 0.2ml / s, and the droplet diameter is stabilized at about 1.0mm. The temperature and humidity control device controls the ambient temperature at 28℃, the relative humidity at 45%, and the film drop height is set to 15cm. During the film drop process, relevant parameters are monitored in real time to ensure stable film production quality.

[0089] (V) Data monitoring and analysis

[0090] The cell genetic information management system collects experimental data in real time and transmits it to the LIS / HIS system in TXT file format. The experimental data were analyzed and the average cell survival rate was 88%, the standard deviation was 1.5%, and the coefficient of variation was 1.7%; the average chromosome dispersion was 8.4, the standard deviation was 0.2, and the coefficient of variation was 2.38%; the average chromosome aberration rate was 2.3%, the standard deviation was 0.5%, and the coefficient of variation was 21.74%. The experimental data was stable and reliable, meeting the expected requirements.

[0091] 4. Comparative Proportion

[0092] 1. Experimental preparation

[0093] 192 samples were processed using traditional manual operation. The culture medium was manually dispensed and the samples were inoculated using an ordinary pipette. The inoculation volume was difficult to accurately control. After multiple measurements, the inoculation volume fluctuated between 0.2-0.6 ml. Due to the accuracy limitations of the pipette itself and the errors in manual operation, the inoculation volume of each sample varied greatly, and the consistency of experimental conditions could not be guaranteed. At the same time, manual operation was inefficient, and it took a lot of time to complete the inoculation of 192 samples.

[0094] (II) Cell culture

[0095] When the inoculated samples were placed in a common incubator, the culture temperature could not be precisely controlled and fluctuated between 35-39°C. Ordinary incubators are not equipped with high-precision temperature regulators, making it difficult to maintain a stable culture temperature. The carbon dioxide concentration could not be stably maintained at 5%, but varied between 3% and 7%. The unstable temperature and carbon dioxide concentration environment had an adverse effect on cell growth, resulting in inconsistent cell growth rates and abnormal morphology in some cells.

[0096] (III) Cell harvesting

[0097] The cell harvesting process uses traditional methods and does not use electric field-assisted cell separation technology. The centrifugation operation uses an ordinary centrifuge, and the speed and time are difficult to accurately control. During the centrifugation process, due to the unstable speed, the cell sedimentation effect is poor and some cells are damaged. The hypotonic treatment and fixation processes rely on manual experience, and the amount of fixative added and the fixation time cannot be accurately controlled. Sometimes too much fixative is added, resulting in excessive fixation of cells and changes in morphology; sometimes too little, the fixation effect is poor and cells are easily lost. It is also difficult to ensure consistent fixation time, which makes the cell fixation effect vary greatly between different samples. 10 samples were randomly selected to test cell-related indicators. The results are shown in Table 6 below:

[0098] Table 6:

[0099]

[0100] Compared with the examples, the cell purity, integrity and harvest volume are significantly lower. Low cell purity means more impurities, which will interfere with subsequent chromosome analysis; poor cell integrity may lead to incomplete chromosome morphology, affecting the accuracy of the analysis results; and low harvest volume may not meet the experimental requirements for the number of cells.

[0101] (IV) Drops

[0102] When using a simple dropper device for slide preparation, it is impossible to control the dropper flow rate, droplet size, and ambient temperature and humidity. During the dropper process, the droplet sizes are different and the flow rate is unstable, resulting in uneven distribution of cells on the slide. In some areas, the cells are too dense and overlap with each other, making it difficult to clearly observe the chromosomes; in some areas, the cells are sparse or even absent. At the same time, due to the lack of temperature and humidity control, environmental factors have a great impact on the quality of the slide preparation. When the humidity is high, the slide is easily damp and the cells are easily deformed; when the humidity is low, the cells are easy to dry, which will also affect the morphology of the chromosomes. The poor preparation effect seriously affects the subsequent chromosome analysis.

[0103] (V) Data monitoring and analysis

[0104] Experimental data relies on manual recording and lacks systematic data processing and quality control algorithms. During the experiment, key indicators cannot be monitored in real time, and abnormal experimental data are difficult to detect and handle. For example, during the cell culture stage, abnormal fluctuations in temperature and carbon dioxide concentration cannot be discovered in time; during the cell harvesting stage, it is impossible to accurately judge whether the cell purity, integrity and harvest volume meet the requirements. Manual data recording is also prone to errors, resulting in low data accuracy and reliability. After the experiment, the data can only be analyzed through simple statistics, and it is impossible to deeply explore the information behind the data, making it difficult to evaluate the stability and reliability of the experiment.

[0105] This study conducted a comprehensive comparative evaluation of the new chromosome analysis smear processing system and the traditional manual processing method through Examples 1-3 and comparative examples. In the experimental preparation stage, the new system uses a high-precision filling pump and automated inoculation equipment to control the culture medium filling accuracy within ±0.15ml and the inoculation volume accurately between 0.3-0.5ml; while the traditional manual operation uses an ordinary pipette, and the inoculation volume fluctuates between 0.2-0.6ml, with large errors and low efficiency.

[0106] In the cell culture stage, the constant temperature culture unit of the new system uses a PID temperature control circuit and a gas concentration control device to maintain a stable temperature at 37°C ± 0.5°C and a carbon dioxide concentration at 5%. The traditional method uses an ordinary incubator with a temperature of 35-39°C and a carbon dioxide concentration fluctuating between 3% and 7%, which cannot provide a stable environment for cell growth.

[0107] When harvesting cells, the new system uses electric field-assisted cell separation technology, combined with precise centrifugation, hypotonicity and fixation processes, to achieve excellent cell purity, integrity and harvest yield. For example, in Example 1, the cell purity reached 92%, integrity 90%, and harvest yield 3.5×10 6 Due to the lack of effective technical means and imprecise operation, the traditional method has significantly low cell-related indicators, with cell purity of only 80%-85%, integrity of 73%-78%, and harvest volume of 2.0-2.8×10 6 indivual.

[0108] During the automatic drop film process, the continuous drop film mechanical structure of the new system can accurately control the drop film flow rate and droplet size, and the temperature and humidity control device stabilizes the ambient temperature and humidity within an appropriate range, resulting in high production quality. The traditional drop film device cannot control the relevant parameters, resulting in poor production effects and uneven cell distribution.

[0109] In terms of data monitoring and analysis, the cell genetic information management system of the new system can transmit data in real time and perform quality control based on threshold judgment and statistical analysis algorithms; the traditional method relies on manual recording and simple analysis, which cannot detect abnormalities in a timely manner, and the data accuracy and reliability are low.

[0110] In general, the new chromosome analysis smear processing system has shown significant advantages in all experimental links, can effectively improve experimental efficiency and result accuracy, and provides more reliable technical support for chromosome analysis.

[0111] Table 7 Summary

[0112] Table 7:

[0113]

[0114]

[0115] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the technical principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A novel chromosome analysis smear processing system, characterized in that: include: Automatic cell inoculation and culture module, automatic cell harvesting module, automatic droplet module, cell genetic information management system; The system is based on a fully automated sample processing platform, with a single batch processing throughput of no less than 192 samples, and the throughput is scalable; The automatic cell inoculation and culture module is used for culture medium packaging, sample inoculation and cell culture; The automatic cell harvesting module integrates multiple cell harvesting processes; The automatic film drop module is used for film making; The cell genetic information management system connects to external systems and monitors data throughout the entire process.

2. A novel chromosome analysis smear processing system as claimed in claim 1, characterized in that: The automatic cell inoculation and culture module is equipped with a high-precision filling pump and a constant temperature culture unit. The culture medium filling accuracy reaches ±0.15ml, the inoculation volume ranges from 0.3-0.5ml, the culture temperature is controlled at 37°C ±0.5°C, and the carbon dioxide concentration in the culture environment is controlled at 5%.

3. A novel chromosome analysis smear processing system as claimed in claim 2, characterized in that: The high-precision filling pump achieves high-precision filling through volumetric quantitative filling technology, which uses a high-precision piston pump structure to accurately control the filling volume of the culture medium by controlling the displacement of the piston under the precise drive of the motor; The constant temperature culture unit uses a PID temperature control circuit combined with a heating wire and a cooling plate to maintain a constant temperature. The temperature sensor monitors the culture environment temperature in real time and feeds the signal back to the PID temperature control circuit, which adjusts the working status of the heating wire and the cooling plate.

4. A novel chromosome analysis smear processing system as claimed in claim 1, characterized in that: The automatic cell harvesting module integrates centrifugation, hypotonic treatment and fixation processes, and adopts two fixation processes, with 6 ml of fixative added each time and a fixation time of 20-30 minutes.

5. A novel chromosome analysis smear processing system as claimed in claim 4, characterized in that: The innovative harvesting method of the automatic cell harvesting module eliminates uncertainties through electric field-assisted cell separation technology, specifically: During centrifugation, an alternating electric field with a frequency of 20-50 Hz and a strength of 0.5-2 V / cm was applied; The centrifugal speed was 1800 rpm and the centrifugal time was 6 min; At the same time, during the hypotonic treatment and fixation process, the presence of the electric field promotes the contact between chemical reagents and cells.

6. A novel chromosome analysis smear processing system as claimed in claim 1, characterized in that: The automatic film dropping module adopts a continuous film dropping mechanical structure and a temperature and humidity control device, the temperature and humidity control accuracy is ±0.25°C and ±2%RH respectively, and the film dropping height is 10-30cm.

7. A novel chromosome analysis smear processing system as claimed in claim 6, characterized in that: The dripper of the continuous dripping mechanical structure adopts a quartz capillary micro-dripper; The driving mechanism is piezoelectric ceramic drive, which applies a changing voltage to the piezoelectric ceramic to cause it to deform slightly, pushing the liquid out of the dripper; By optimizing the inner diameter, length and shape of the internal flow channel of the dripper and utilizing Poiseuille's law in fluid mechanics, the flow rate and droplet size of the drip can be precisely controlled.

8. A novel chromosome analysis smear processing system as claimed in claim 1, characterized in that: The cell genetic information management system transmits data to the LIS / HIS system in real time, and internally uses an algorithm based on threshold judgment and statistical analysis for data processing and quality control; The specific threshold judgment rules are as follows: the normal threshold range of cell survival rate is set to 70%-95%, the normal range of chromosome dispersion is 7-9, and the normal threshold range of chromosome aberration rate is 0-3%; when the cell survival rate is lower than 70% or higher than 95%, the chromosome dispersion is lower than 7 or higher than 9, and the chromosome aberration rate is higher than 5%, the system automatically determines that the data is abnormal and triggers an early warning; The algorithm of statistical analysis is as follows: collect the cell survival rate, chromosome dispersion and aberration rate in each batch of experimental data, calculate the mean value, standard deviation and coefficient of variation of each indicator, and if the coefficient of variation of an indicator is greater than 10%, it is judged that the stability of the experimental data of this batch is poor, and the system prompts the operator to check the experimental equipment or re-run the experiment; If the average value of a certain indicator of consecutive specified batches exceeds the normal threshold range, the system will also issue an early warning and prompt relevant inspections and adjustments.

9. A novel chromosome analysis smear processing method, characterized in that: The implementation of the method is based on a novel chromosome analysis smear processing system as described in any one of claims 1 to 8, comprising the following steps: Step 1: Use the high-precision dispensing pump in the automatic cell inoculation and culture module to automatically dispense the culture medium, and use the micro-injection pump combined with a high-precision flow sensor to control the inoculation volume within the range of 0.3-0.5 ml for sample inoculation; Step 2: Place the inoculated sample in the constant temperature culture unit of the automatic cell inoculation and culture module, use the PID temperature control circuit combined with the heating wire and the cooling sheet to maintain the culture temperature at 37°C ± 0.5°C, and use the gas concentration control device to maintain the carbon dioxide concentration in the culture environment at 5% for constant temperature culture; Step 3: Through the automatic cell harvesting module, electric field-assisted cell separation technology is used to first perform centrifugation operation, and centrifuge at a speed of 1800 rpm for 6 minutes under an alternating electric field environment with a frequency of 20-50 Hz and an intensity of 0.5-2 V / cm to allow the cells to initially settle; then add 0.075 mol / L potassium chloride as a hypotonic solution for hypotonic treatment, and the treatment time is 10 minutes; finally, two fixation processes are performed, each time the amount of fixative added is 6 ml, and the fixation time is 20-30 minutes, and the cell harvest is completed; Step 4: Use the continuous film drop mechanical structure and temperature and humidity control device of the automatic film drop module to make films. The piezoelectric ceramic drives the quartz capillary micro-droplet head to achieve continuous film drop. The film drop flow rate is controlled to 0.2ml / s, and the droplet diameter is controlled to 1.0mm±0.05mm. At the same time, the temperature and humidity are controlled at 20-40℃ and 25%-75%RH respectively by using the temperature and humidity control device combined with the compressor and the semiconductor. The height of the film drop is 10-30cm; Step 5: Through the cell genetic information management system, using an algorithm based on threshold judgment and statistical analysis, upload the data during the experiment in real time, and monitor the quality control parameters of cell survival rate, chromosome dispersion and aberration rate. When the data is abnormal, timely warnings will be issued and the corresponding processing flow will be initiated.

10. A novel chromosome analysis smear processing method as claimed in claim 9, characterized in that: In step five, if the cell survival rate is lower than 70%, the system prompts to check whether the culture conditions are abnormal; if the chromosome dispersion is lower than level 7, it prompts to optimize the droplet operation parameters; if the chromosome aberration rate is higher than 3%, it prompts to check whether the experimental samples are contaminated or whether the experimental process is wrong.

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