Automated robot for in vitro freezing of germ cells and embryos

Abstract

The process of freezing mammalian gametes and embryos has always been known as a very slow process. So far, no device has been developed that is easy to use and usable by non-professionals and can automatically conduct the cell freezing process. In conventional freezing methods, due to the time constraints that are considered for placing samples at predetermined intervals on nitrogen vapor, it is not possible to arrange a large number of pivots simultaneously because moving these pivots from the previous distance to the next stopping distance is time-consuming and the time difference created between the first and last pivots that are moved affects the result and efficiency of freezing. The application of the present device is to perform simultaneous freezing of more than 500 samples in one cycle without any fluctuation between the samples.

Description

Description of the invention Title of the invention (as stated in the declaration) Automated robot for in vitro freezing of gametes and embryos Automated Robot for Gametes and Embryos Vitrification Inventor / Inventors: Navid Dadashpour Devachi, national code 0065790561, born on 05 / 17 / 1988, issued in Tehran, ID card number 4370, father's name Ali Akbar Seyyed Javad Hosseini, national code 0322607353, born on 11 / 12 / 1360, issued from Karaj, ID card number 1582, father's name Seyyed Hossein Technical background of the relevant invention The technical field of this invention is related to the field of automated cell freezing devices, and in particular, it can automatically conduct the freezing operation using only liquid nitrogen vapor, without the need for the complexities of programmable freezers, and without the intervention of human force during the vitrification process. Technical problem and stating the objectives of the invention The process of freezing mammalian gametes and embryos has always been known as a time-consuming and very slow process, both in the research and treatment sectors. The process of freezing gametes in glass, because it has always been done manually and using basic equipment and manpower, has led to increased pressure and workload and shortage of manpower on the days when this process must be done in the laboratory. So far, a device that is easy to use and usable by personnel without scientific and professional expertise and can automatically guide the cell freezing process has not been built. Therefore, after examining and in order to save time and costs of specialized manpower in modern reproductive technology laboratories and fertility and infertility clinics, I designed and built an autonomous robot for the glass freezing of gametes and embryos for the first time in the world. Description of the state of the prior art and the history of developments related to the claimed invention. So far, only one device has been designed in this field in the world. Its focus has been more on the process of filling the pivot and freezing straws with samples and freezing solutions, and its mechanism is based on the very expensive mechanism and technology of programmable freezers with the ability to reduce the temperature to -180 degrees Celsius, which are very expensive and their provision for research and medical centers is not economically justified. In patent number United States Patent 9228925 on 01 / 05 / 2016, the University of Kansas (Lawrence, KS, US) disclosed a device in which loading of cell samples and freezing are performed automatically and based on the technical knowledge of programmable freezers. Providing a solution to an existing technical problem along with an accurate, sufficient, and integrated description of the invention Vitrification of cells is considered the most up-to-date and effective method of freezing oocytes, sperm, and embryos (regardless of the specific stage of development). When we talk to embryologists and people working in fertility and infertility clinics, they all point out that vitrification of gametes and embryos is a time-consuming and very slow procedure. In carrying out this process, careful training and, of course, the accuracy and patience of the relevant technician can have a great impact on the expected efficiency and output of the work. Apart from careful training and considering the necessary accuracy and patience in this process, it should be kept in mind that on the days when the freezing program is in place in the laboratory, the workload increases greatly and the laboratory faces a shortage of human resources in the process.Also, in the usual methods that use nitrogen vapor and liquid nitrogen to freeze pivots and straws containing gametes and embryos, due to the time limit for placing the samples at predetermined distances on the nitrogen vapor, it is not possible to arrange a large number of pivots, because moving these pivots from the previous distance to the next stopping distance is time-consuming, and the time difference between the first and last pivots that are moved can affect the result and efficiency of freezing. Therefore, having a device that is at the lowest cost, unlike foreign models that have very expensive prices and complex mechanisms, and also with very easy-to-use capabilities that reduce the need for expert personnel to zero and the ability to automate the glass freezing process with the help of nitrogen vapor and liquid nitrogen from the beginning to the end of the procedure and with a performance accuracy of 0.01 seconds, can be desirable for all laboratories and users of such a technique in the field of knowledge of modern reproductive technologies.Using this device, the accuracy, quality and quantity of the vitrification process of cells is improved. It is important to note that one of the advantages of this device is the simultaneous entry and exit of samples at a certain height from the nitrogen surface up to 500 samples per second. Also, more importantly, in the process of moving the samples, since this movement is completely robotic, there is no difference in the speed of movement or accidental exit of the sample from the nitrogen vapor. Explanation of shapes, maps and diagrams The technical drawing of the device includes the following. Power 12 V: It is used to power the main 12V relay. Power 7.9 V: It is used to power two device modules. Power 5V: It is used to move the lift motor. Module number one: First relay: Normally closed. After pressing the start button, it opens and causes the lift to move downward by supplying negative current from the 12V power supply to the main relay. When given to this relay on a tenth of a second scale, it determines the amount of motor movement and lift displacement. When the countdown of the first relay ends, the current is cut off, the lift stops, and the countdown of the second relay is activated. Second relay: The function of this relay is to determine the first stop time. Third relay: Like the first relay, it is responsible for moving the lift downwards. Fourth relay: This relay issues a command to start the second module by establishing a short circuit. Module number two: First relay: is responsible for counting down the duration of the second stop. Second relay: Same as the first relay of the first module Third relay: It is responsible for counting down the duration of the third stop. Fourth relay: Like the first relay, the first module, by connecting the current, causes the lift to move downwards, and this movement will continue until the test tubes are completely immersed in nitrogen, which will finally be completely turned off at the end of this stage through the bottom microswitch of the device. 3-position switch: Up: In this position, the lift moves up and automatically stops when it reaches the highest point to start work. Middle: In this state, the lift does not receive any commands from the modules. Down: In this state, the device is ready to start working. Upper NO microswitch: It is responsible for cutting off the power to the lift when the lift reaches its highest height. NO-Lower microswitch: It is responsible for cutting off power to the modules when the lift reaches its lowest level. Relay: It is responsible for connecting the current from the module to the motor and moving the lift. Start key: After programming the modules and placing the status key in the down position, pressing this button starts the device. A clear and precise statement of the advantages of the claimed invention over prior inventions. 1- Very low cost of manufacturing the device, 2- Very simple programmability and use, 3- Possibility of loading 500 samples simultaneously, which has not been reported in any device, 4- Performing the freezing procedure using a vapor temperature gradient on the nitrogen surface, 4- Not using the complex mechanisms of common programmable freezers that have very high prices. Description of at least one implementation method for implementing the invention In order to freeze monkey sperm, according to the available instructions, we program the first stop distance from the liquid nitrogen surface to 10 cm via the keyboard, set the duration of this stop to 10 minutes, and then set the next stop at a distance of 7 cm from the nitrogen surface via the keyboard, and set the duration of the stop to 20 minutes. Then we transfer the samples to a special chest that is connected to the device lift and press the start button. From now on, all steps until the samples are immersed in nitrogen are performed automatically and without the need for human presence. And at the end of the work, the device automatically turns off. At this time, the samples have been completely placed in liquid nitrogen, maintaining the standard procedure. Express mention of the industrial application of the invention Use in embryology laboratories and fertility and infertility centers Brief description of the invention Automated robot for in vitro freezing of gametes and embryos The freezing process of mammalian germ cells and embryos has always been known as a very slow process. So far, a device that is easy to use and usable by non-professionals and can automatically guide the cell freezing process has not been developed. In conventional freezing methods, due to the time limit for placing samples at predetermined distances on nitrogen vapor, it is not possible to arrange a large number of pivots simultaneously, because moving these pivots from the previous distance to the next stopping distance is time-consuming, and the time difference between the first and last pivots that are moved affects the result and efficiency of freezing. The application of the present device is to perform simultaneous freezing of more than 500 samples in one cycle and without any oscillation between the samples.

Claims

Claim What is claimed: Claim 1) A lifting device is connected to two programmable time controllers that regulate and control the values ​​of the stop periods and stop intervals from the surface of the liquid nitrogen in the device's tank. Claim 2) According to claim number 1, the accuracy of the lift movement is within 0.01 seconds. Claim 3) According to claim number 1 and claim number 2, the variance in the exposure times of samples processed on different days and times of processing is equal to zero. Claim 4) According to claim 1 and claim 2, the time / Brodt gradient is available in the temperature range of -20°C to -196°C and in the time range of 0.01±1.

Images