TR202007738A2 - CELL SEPARATION DEVICE BASED ON MAGNETIC LEVITATION PRINCIPLE ON MICROfluid Chip - Google Patents

Abstract

The invention relates to a microfluidic device that enables the separation of circulating tumor cells from white blood cells with the help of magnetic forces without using the labeling method. With the invention consisting of 4 mirrors, 2 magnets, separator, a special channel design with one inlet and two outlets, the cells mixed with the paramagnetic solution are directed to different heights according to their density and can be collected by being separated under flow. Without changing the magnet positions and the concentration of the paramagnetic solution, the separable density range can be changed simply by moving the microfluidic channel in the horizontal axis. The harvested cells can be used in molecular testing to diagnose cancer or monitor treatment success.

Description

DESCRIPTION PRINCIPLE OF MAGNETIC LEVITATION ON MICROfluidics Chip Technical Field of the Invention The invention does not use the labeling method of circulating tumor cells from white blood cells. It is related to the microfluidic device used for separation with the help of magnetic forces.

State of the Art of the Invention (Prior Art) Circulating Tumor Cells (STH) are both present in very small numbers in the blood. (such as 1 STH per 1 million white blood cells), as well as in the context of biomarker (biomarker) They are very difficult to isolate because they are heterogeneous. Circulating circulating tumor There is no specific marker that definitively distinguishes blood cells from blood cells. Different Tumors of histological and molecular types express different marker sequences and Even the histological tumor type shows great immunologic and molecular differences. in question.

Current STH extraction methods, size between cells for this process, electrical charge, consists of techniques that utilize differences in density and cell membrane antigens.

However, these techniques require manual preparation of samples, low separation cost. difficulty and efficiency, such as causing inconsistent results and high costs can lead to troubles. They are both very low in blood and heterogeneous. Since they have biomarkers, they have been effective and efficient in separating STHs so far. A standard method has not been developed. Taking advantage of its difference, cells in a heterogeneous cell community are placed in the capillary channel. due to the difference in the positions where the magnetic force and gravitational forces are balanced. A system that can separate using Levitation mentioned in the document system, a microscope on which a microcapillary channel is placed, two neodymium (NdFeB) magnet and two mirrors are used. Cells and magnetically active agent (gadolinium-based, non-ionic paramagnetic medium), a glass microcapillary It was loaded into the tube and placed between two opposite magnets and the cells were in a static state. has been observed. However, since this system does not take place under flow, it is necessary to collect cells. It does not provide an opportunity, it only allows observing the positions of the cells. decomposition of heterogeneous populations in a microfluidic environment with one or more magnets It is related to. Magnetic integrated with the mobile phone camera mentioned in the document The levitation platform is mostly used for counting blood cells; blood count, to observe the development of the disease and the effect of the drug on the disease. is being done. In the patent in question, analysis is performed by looking at cells in a glass capillary tube. is being done. In addition, the plastic design shown in the patent, while separating the cells It does not allow to be observed, which allows the determination of cell density by separation. does not allow. The plastic channel used in the aforementioned patent is a double layer of three plastic plates. It is produced by sticking the sides together with adhesive tapes, and the thick and rough channel Its walls do not allow cells to be observed between magnets. Pine Since the capillary tube is transparent, it allows viewing, but the glass tube is a single displayed in different positions in the channel due to the fact that there is only one entry and one exit point. cells cannot be collected from different exit points and also a flow of the glass tube will be applied. It has the disadvantages of not having a design.

Separating STHs from white blood cells without marking and collecting the separated cells on the subject, the samples were not prepared manually, the separation purity was high and a standard method was used. method development is needed.

Brief Description of the Invention The invention is aimed at the early diagnosis, prediction and personalized treatment of cancer. circulating tumor cells, which play a vital role in determining that provides the separation and makes this separation with the magnetic levitation principle. presents a microfluidic chip. The aforementioned invention has been used in the last period of cells and cellular very successful in the biological characterization and observation of activities. a new method based on the magnetic levitation principle, which has emerged as a to address this need for the segregation and collection of STHs by developing aims.

With the developed microfluidic chip, the white blood cells of STHis are removed at high speed. High rate of separation in as little as one hour thanks to the flow (1 mL/hour) is targeted. Cell separation platform based on the invention magnetic levitation principle, 1 mL/hour (~15 µL/min) from the cells in the microfluidic channel with the help of a syringe pump. By giving the flow, it is ensured that it is separated under a continuous flow. Decomposition in the invention It is dynamic and at the end of the 1 hour period, the cells go to different outlets with the flow given to the channel. can be directed. separation and collection of cancer cells from a heterogeneous environment without blood analysis is on it. In the inventive system, unlike the patent documents in the previous art, The channel in which the cells are located is not a glass tube or plastic channel, but the channel wall. Flow-resistant and discrimination, where distances and separator point position can be controlled with the advantage of being transparent, having different channel exit points after A special microfluidic channel design is presented, which makes it possible to

No prior art papers indicate that STH°s are labeled from white blood cells, decompose under continuous flow by magnetic levitation in the microfluidic chip It is seen that it does not offer a system that provides

In the invention design, there is no use of capillary glass, instead one entrance and two exits. A microfluidic channel (1) made of glass and polymer with a dotted to visualize cells and analyze their density, and also to allows for differentiation. In the invention, cells with different density are different. It is collected by the effect of flow from the exits and thus cancer cells are removed from blood cells. It can be separated and is a preliminary step for the next diagnostic and therapeutic analysis. is happening.

Also, in the device design presented in the invention, there is no magnet position and paramagnetic without changing the concentration of the solution, simply by shifting the channel (l) on the horizontal axis The separable density range for cells can be changed. In this way, only one possible to separate different cells and particles using microfluidic channel (1) design is locked.

The microfluidic channel (1) used in the invention system is privately owned by the people to whom the invention belongs. is designed as The glass capillary tube used in the prior art is standard with certain dimensions. It is a tube and it is not possible to change the dimensions of the channel width or length. It is not suitable for use in cell differentiation experiments. For this reason, in the desired width in the invention, It is made of glass (1 1) and polymer, where the wall thicknesses and the position of the separator (12) can be controlled. A fabricated microfluidic channel is designed. The aforementioned polymer is based on silicone. It can be elastic. In the invention, polydimethylsiloxane, which is preferably one of the silicone-based elastomers (PDMS) is used.

In the inventive system produced to separate cancer cells from blood cells, the microfluidic channel (1) soft lithography with high resolution, desired channel width and reagent (12) by producing the mold with position 1 and using polydimethylsiloxane (double-agent polymer). amplified and then adhered to the glass (1 1) surface with oxygen plasma.

There is a glass (11) surface on the lower and upper surfaces of the microfluidic channel (1).

In the invention microfluidic chip, four mirrors (6) placed at an angle of 45° and the microfluidic With the advantage of the transparency of the inside of the channel (1), the cells can both be separated under flow and It is collected at different outputs and can be displayed simultaneously. upper channel and 2 neodymium magnets (4,5) with opposite poles crossing each other are placed under it.

Thus, a channel design that is thin enough to enter between two magnets is provided. The thin glass (1 l) surface allows visualization of cells.

The inventive device can be used to detect magnetic forces without using the marking method of cancer cells. It is the first product to demonstrate microfluidic on-chip decomposition with the help of has. In addition, with the proposed invention and method, it is possible to separate the cancer cells from the blood cells. instead of complex, expensive devices that require specially trained personnel to use. usability, independent of the marking technique, therefore inexpensive platform can be designed and development is provided.

The most important goal of the invention is to differentiate cancer cells without being labeled with antibodies. This Thanks to this feature, cancer cells are independent of the antigens on their surface. can be differentiated. Same type of cancer cells have different surface antigens Considering that, the recommended method is to blot a larger cancer cell population. It has the feature of separating it from its cells.

The magnetic levitation principle used in the invention is to separate cells from each other under flow. It has never been used before for collecting. In this context, thanks to the invention, the necessary infrastructure With the acquisition of this technology, it also contributes to the applications of this technology in different fields. will be found. The invention is aimed at diagnosing and improving the course of cancer by distinguishing STHs in the blood. can be used for tracking. In addition, the separated STH71ers are more effective with cancer. also in the development of personalized treatment methods to combat they can be used.

The invention will also allow the development of different cell sorting projects.

Among these studies; without being marked in the blood of microorganisms such as viruses, fungi and yeast identifying and separating them and administering the treatment specific to the microorganism found to the patient; isolating drug-resistant microorganisms and producing more effective drugs for them. development; pregnant as an alternative to the amniocentesis method, which carries significant risks. Prenatal genetic tests by separating fetal cells in women's circulation It is performed without risk in the separated cells.

Description of Figures Explaining the Invention Figure 1: Visualization of the forces acting on the cells on the magnetic levitation platform.

Figure 2: Magnetic magnet consisting of two magnets, four mirrors and a PDMS microfluidic channel levitation apparatus.

Figure 3: Magnetic levitation in a microfluidic channel under flow of cells with different densities the principle of separation in the arrangement.

Figure 4: A. Microfluidic channel with one inlet and two outlets, B. From the separator point of the channel PDMS upper and lower canal wall widths in the section taken.

Figure 5: Microfluidic chip and magnet positions A. Microfluidic channel with magnets in the same position, B. The microfluidic channel is y=250 µm away from the magnets position, C. The microfluidic channel is y=500 µm away from the magnets D. The microfluidic channel is at a distance of y=1000 µm from the magnets. Figure 6: Density range that can be separated according to magnetic positions in the microfluidic chip. mM) and under different flow rates (5 µL/min, 10 µL/min, 15 µL/min. 20 µL/min) separate data rates.

Figure 8: Aggregation in the upper channel of the U-937 cell line at different cell and Gd concentrations efficiencies.

Figure 9: MDA-MB-231 cell line in the upper channel at different cell and Gd concentrations collection efficiencies.

Definitions of Invention Elements 1. Microfluidics Channel Microfluidics Channel Upper Wall Microfluidics Channel Bottom Wall Top Magnet Bottom Magnet Upper Channel Sub Channel Upper Channel Output . Sub Channel Output . Reagent Detailed Description of the Invention The “sample to be separated”, which is aimed at separating the cells in it, is the invention microfluidic are given to the device and the cells collected at different heights according to their density, under flow. can be collected separately. Collected cells, cancer diagnosis or treatment success It can be used in molecular tests to monitor

The “sample to be separated” referred to in the invention is blood, serum or blood containing different cells. could be a plasma sample. Cells in the sample to be separated mentioned in the invention; virus, microorganisms such as bacteria, yeast, cancer cells, circulating tumor cells, red blood cells, white blood cells, stem cells, and platelets. The phrase "cell solution" defines the sample to be separated when mixed with the paramagnetic solution. Meet separation of all aforementioned cell groups from the cell mixture/sample can be provided.

For this purpose, Polydimethylsiloxane (PDMS) microfluidic chip by soft lithography method. is produced. The upper and lower wall thicknesses of this chip are thin (<800 pm (200-800 pm) by means of the mold. A high magnetic field is obtained in the channel by keeping it between 400-600)) has been made. Keeping the distance (<2 mm) between the upper and lower magnets increases the magnetic field values, which results in a less concentrated paramagnetic solution. allows analysis. Highly concentrated paramagnetic solutions On the other hand, it affects the viability of the cells and this causes the decomposition efficiency to decrease. can happen.

The microfluidic channel has one input and two outputs. A constant thickness in the output section reagent with a long (thinner than the size of the canal) and a pointed end, passes through the midpoint of the channel, which is the dividing point for cells of different heights. It allows the cells to be directed to different outlets and thus collected from different outlets. it provides. Position of said separator to parse cells/redirect to different outputs in terms of importance. The thickness of the separator, the size of the channel and the cells to be separated can be changed according to its density. The thickness of the separator (12) mentioned in the invention is 25-200 µm range, preferably 100 µm thick.

In one embodiment of the invention, the "sample to be separated" is cancer and blood cells, pump The sample to be separated (cancer and blood cells) given from the canal entrance (13) through the canal Depending on their density, they reach the equilibrium height and cancer cells in the exit part. From the top of the separator, the blood cells are directed to different outlets from the bottom of the separator. Meet It has two strong magnetic properties fixed by the parts surrounding the microfluidic chip. By using neodymium magnet (N52), high magnetic forces can be achieved and high speed (l mL/hour (may be in the range of 0.1-2mL/hour), cancer and blood cells with different density segregation is carried out. In this respect, the invention is for the early diagnosis of cancer. It is an innovative product within the scope of the studies carried out.

The separation provided by the invention is not in static form but is dynamic and at the end of a 1 hour period. cells can be directed to different outlets with the flow given to the channel. As shown in Figure 1 and Figure 4 PDMS microfluidic channel produced by soft lithography method as The wall (2) thicknesses were kept thin (200-800mm) by means of the mold obtained from the three-dimensional printer. um (preferably 400-600)), the upper and lower magnets (4 and 5) more influence into the channel and thus a high magnetic field is obtained in the channel. In this way, each The magnetic force acting on the particle or cell is kept high.

In the magnetic levitation device, microparticles in the paramagnetic solution/ cells move towards the midpoint of the two magnets, which is the point in the channel where the magnetic field is the least. they will move. The middle point of the magnets (as the levitation height) is the lowest point of the separator. Unlike the levitation height at the tip of the spike, after the cells are in equilibrium, then it ensured that they were collected from different outlets depending on their density.

In the magnetic levitation method, cells and the paramagnetic solution in which the cells are mixed negative magnetic susceptibility difference between lifts to heights. This height depends on the density of the cells, but differs from their dimensions. is independent. This method measures cell densities down to the single cell level, while at the same time It is the only place where cells can be observed and identified without the use of a labeling method. method. Using this method, each cell type has a unique levitation, i.e. It can be observed that there is a density profile. Channel between magnets with opposite poles negative magnetic susceptibility between cell and solution in levitation of cells in The difference is that the cells move towards B (magnetic flux density) with small magnetic field strength. it moves. The magnetic force (Fm) acting on the cell is equal to the buoyant force (Fo) of the liquid. At that point, the cell remains motionless. This point that the cell reaches depends on the density of the cell. it is attached.

Magnetic principles: A particle (or a particle) in a liquid under the influence of a magnetic force. the net force (FNet) acting on the cell until it reaches the equilibrium position in the system; magnetic force (FM) is the resultant of drag force (FD) and gravitational force (FG) (equation 1 set). In this system, the inertial force (Fi) is determined by the microfluidics magnetophoresis. Due to the low Reynolds number, the Brownian force (FB) is only small enough. Because it significantly affects the movement of particles (approximately 510 nm), can be ignored.

E-er=îi+î+l~î (1) FM acting on a particle; B: magnetic flux density (Tesla, T), V: del operator, m: it depends on the magnetic dipole values (equation 2). As B moves away from the surface of the magnet is decreasing. From here, in a paramagnetic salt solution or in ferrofluid, low the magnetic dipole produced in the magnetic field can be reached (equation 3).

FM : (NI.V)B (2) and #o (3) Moon: arranged as the magnetic susceptibility difference (Xp-Km) of the particle and the surrounding environment The FM value can be expressed (equation 4).

It is expanded in the (B.V)B Cartesian coordinate system, to the magnetic flux values in three dimensions. accessible. at 68.dB.

BxTI* B}.Ö_`;`+BZÖ_Z" 68 dB Exa-I +B}.Ö_\_Z+ Eta-ZZ If the other force acting on the particle is the hydrostatic buoyant force (FG); V: your particle volume, Ap: volumetric density difference between the particle and the surrounding medium (Pp-PM), g: gravity It can be calculated depending on the function (9.8 in-s_2) (equation 6).

= Vûpy (6) The cell is at rest at the point where the magnetic force is equal to the hydrostatic buoyant force. will stay. However, if a perpendicular flow to the magnetic forces is applied during magnetic levitation, the cells will start to drift (Figure 1). a spherical particle under specified conditions. Drag force FD, which is another force acting on it; R (particle based on radius), n (dynamic viscosity), fd (entrainment coefficient), and vp (particle velocity) will change (equation 7).

FT; : 6nRiifd(vp) (7) Cell lines grown in normal cell culture in tests (example to be separated) to model), the PDMS on the microfluidic chip is connected to the microfluidic channel (1), figure As shown in 4, the input is given from the person (13) with the help of a syringe pump. in Figure 2 As shown, on the invention, magnetic applied to both the PDMS microfluidic channel (1) In order to increase the effect and to ensure that the magnetic effect is collected only on the channel. 2 pieces in the form of rectangular prisms, the upper magnet (4) and the lower magnet (5). neodymium iniknatis was placed. Placed at an angle of 45° where viewing is possible Plastic produced by 3D printer with 4 mirrors (6) and one PDMS microfluidic channel (1) parts are integrated with each other. Being transparent, pressure resistant and soft Since it is the most suitable polymer for the lithography method, PDMS is used as the channel material. polymer was chosen. Said polymer may be a silicone-based elastomer. In the invention, preferably Polydimethylsiloxane (PDMS), one of the silicone-based elastomers, is used. Soft Different flexible and transparent polymers such as Ecoflex compatible with lithography can also be used in the microchannel of the invention. can be used in production.

Before cell transfer, cells contain a determined concentration of gadolinium ion (Gd). It was prepared in fetal bovine serum (F BS) containing the agent. This agent is paramagnetic. and magnetic resonance imaging (MRI) of cancerous tissues with its water-soluble structure. It is a very active substance in diagnosis. The Gd concentration determines the levitation heights of the cells. and it can change the speed of reaching this height. Except for the GD ion, especially in MR Different paramagnetic ions (such as metal manganese ((Mn2+)) can also be used.

The cell separation protocol includes the following steps and is detailed below. described in sections: 1. Final solution in cell solution 30 mM (10-100 mM concentration the sample to be separated, with Gd in concentration is mixed. 2. 1 mL of cell solution (cell solution) containing the sample to be isolated and Gd ions amount is 0. It is drawn into the syringe and pumped into the syringe pump. is placed. 3. The tip of the syringe is connected to the microfluidic chip inlet (13) with the help of a capillary tube, the upper These outlets are used to collect cells separated from the channel outlet (9) and downstream outlet (10). It is connected to two tubes with the help of capillary pipes. 4. A maximum flow of 2 mL/hr (preferably 1 mL/hr) is initiated from the syringe pump and the cell solution is fed into the PDMS microfluidic channel (1).

. After the entire cell solution is delivered to the PDMS microfluidic channel (1), the upper channel The dissociated cells are collected from the tubes connected to the outlet (9) and the lower channel outlet (10).

Optimization of Gd Concentrations and Cell Delivery Rates In an application of the invention. to effectively separate cancer cells from white blood cells. cell delivery rates and Gadavist concentration are optimized. For this microparticles close to the density of these cells instead of cancer cells and white blood cells used. Microparticles are in the invention only for optimization, cancer cell and white It is used to mimic blood cells. solution containing cells with PDMS microfluidic channel (1) with a syringe pump. When administered, they have a lower density (1.06 g/mL) than cells advancing in the downstream direction. cells will be collected from the upper channel outlet (9) of the mold channel on the separator (12). More Since cells with high Density (1.09 g/mL) remain in the lower part of the separator (12), the lower channel They will be collected from the outlet (10).

In this direction, the movement and separation of the cells within the magnetic levitation mechanism. to two different cores with diameters of 10 am and 20 am to monitor and optimize the system. (1.02 g/mL and 1.09 g/mL) particles were used and different Gadavist pL/min, 20 µL/min) dissociation efficiencies in phosphate buffered saline (PBS) calculated (Figure 7). Microparticle with separation efficiency collected from the upper channel outlet It is calculated as the ratio of the number of microparticles collected from the lower and upper channel outlets.

Flow rate and Gd concentration are factors affecting separation efficiency. Namely; Gd Changing the concentration changes the final levitation height of the particles and Increasing the flow rate will separate the particles before they reach the final levitation height. causes it to reach. Gd concentration and flow giving separation efficiency above 90% speed range has been determined. Microparticles with a density of 1.02 g/mL 15 µL/min and 30 Mm Gd It was collected from the upper channel outlet with a separation efficiency of 93.11% in the containing medium. same conditions 14.13% of particles with a density of 1.09 g/mL below were collected from the upper channel.

Microparticles fetal bovine serum to model patient blood (sample to be separated) It is prepared in medium containing (PBS) and 30 mM (10-100 mM concentration) in the solution. It can be in the range of the microfluidic channel Gd is given to the channel. Gd added in The solution has become paramagnetic and has a density of 1.02 g/mL and 1.09 g/mL. microparticles from the upper channel at ~90% and 5% efficiency, respectively, under 15 µL/min flow. has been collected. The flow rate at which the results are obtained (15 µL /min) corresponds to the desired flow rate (1 mL/hour) is close. Also, microparticle separation efficiencies in PBS and FBS is similar.

Three different concentrations of white blood to model patient blood (the sample to be separated) cells/mL) at different concentrations of Gd (20 mM, 30 mM and 40 mM) 1 mL/h flow and their efficiencies were calculated (Figure 8-9). Separation of microparticles The optimum Gd concentration selected as a result of the results was consistent with the cell sorting experiments. has been observed. Likewise, in 30 mM Gd, at a concentration of 103 MBA-MB-ZSl/mL A separation efficiency of 66.75% was observed at a concentration of 66.75%.

In addition, realizing the separation efficiency of over 901% with the invention, that is, white blood in order to collect almost all of the cells from the lower canal, the lower part of the white blood cells Increasing magnet pulls could be the next goal. For this reason, these cells magnetically coated with CD45 antibody that will enable them to bind to antigens on its surface. higher than the lower channel outlet (10) of white blood cells labeled with nanoparticles can be collected efficiently. Namely; CD45 binds to white blood cells and will change the magnetic susceptibility of the white blood cells and thus the lower channel of the white blood cells. The difference in levitation height between cancer cells by ensuring that they are collected at the level of (8) will increase. This will increase the separation efficiency.

As a result of the above-mentioned optimizations, the Gd concentration suitable for separation The rate of delivery of mM and cell solution to the canal was determined as a maximum of 1 ml/hour.

However, when separating with the invention, the Gd concentration is in the range of 10-100 mM and the cell The rate of delivery of the solution to the canal can be in the range of 0.1-2 ml / hour. However, different Gd If it is desired to work with the same efficiency in the concentrations of the magnets to each other, positions can be changed.

As shown in Figure 51, the microfluidic chip can be formed in different shapes according to the magnets. positioned inside the channel without changing the amount of Gd and the distance between the magnets. The separable density value can be changed. of the microfluidic chip to the magnets As the distance increases on the horizontal axis, the magnetic effect on the microparticles/cells the force is changing, which means that the microparticles/cells flow into different Output channels. influences its orientation. In cell sorting experiments, PDMS microfluidic It is positioned so that the horizontal distance of the channel (l) to the inductions is y:0 µm (Fig. A.). The distances of the magnets to each other are kept constant and only the microfluidic chip y=250 µm (Figure 5 B.), y=500 µm (Figure 5 C.) in the horizontal direction from the outer frame of the magnets and When y=1000 µm (Figure 5D.) is removed, the separable density value decreases. observed according to the simulation results (Figure 6). Thus, the same microfluidic chip using the relative position of the magnets and the paramagnetic solution. without changing the concentration of the microfluidic chip only horizontally relative to the magnets. By changing its position on the axis, cells in different density ranges can be separated.

Claims (1)

REQUESTS It is a method of separating cells by magnetic levitation and its feature is, i. Creating a cell solution by mixing the sample to be separated with a paramagnetic solution, ii. Aspirating the cell solution into the syringe and placing it in the syringe pump, iii. Connecting the tip of the syringe to the microfluidic channel inlet (13) with the help of a capillary tube, iv. Placement of the upper magnet (4) and the lower magnet (5), with opposite poles facing each other, above and below the microfluidic channel (1). ,10) connecting two tubes with the help of capillary pipes, vi. Starting the flow from the syringe pump and giving the cell solution to the microfluidic channel (l) under flow, vii. After the entire cell solution is given to the microfluidic channel (1) under flow, the cells separated by magnetic levitation in the microfluidic channel (1) between the upper magnet (4) and the lower magnet (5), the opposite poles of which are facing each other, the upper channel outlet (9) and the lower magnet (5). It is collected from the tubes connected to the channel outlet (10) and includes the processing steps. . It is a method according to claim 1. Its feature is that the sample to be separated is a blood sample, serum sample, plasma sample or cell mixture sample containing fetal bovine serum (FBS). It is a method according to claim 1, characterized in that the cells in the sample to be separated are viruses, bacteria, yeast, cancer cells, circulating tumor cells, red blood cells, white blood cells, stem cells or platelets. It is the method according to claim 1, characterized in that the concentration of the said paramagnetic solution is in the range of 10-100 mM in the cell solution. The method according to claim 4, characterized in that the concentration of said paramagnetic solution is 30 mM in the cell solution. A method according to any one of claims 1, 4 or 51, characterized in that said paramagnetic solution is a solution containing gadolinium (Gd). It is the method according to claim 1, characterized in that the amount of said cell solution is in the range of 0.1-10 1nL. It is a method according to claim 7, characterized in that the amount of said cell solution is 1 mL. It is the method according to any of the claims 1, 7 or 8, characterized in that the delivery rate of the said cell solution to the channel is in the range of 0.1-2 m1/hour. It is a method according to claim 9, characterized in that the delivery rate of the said cell solution to the channel is 1 ml/hour. It is a method according to demand, and its feature is that the material of the mentioned microfluidic channel (1) is silicon-based elastomer. It is a method according to claim 11, characterized in that the said silicone-based elastomer is polydimethylsiloxane (PDMS). It is a device for separating cells with magnetic levitation, and its feature is that it works according to the 1" request. It is a device according to claim 13 and its feature is, i. It consists of an inlet (13), the upper channel (7) and the lower channel (8), separated by a separator (12) inside, the upper channel outlet (9) and the lower channel outlet (10) at the end of these channels, where the capillary pipes where the cells will be collected are connected. microfluidic channel (1) with two outlets, ii. 2 magnets, the upper magnet (4) and the lower magnet (5), whose opposite poles are facing each other, placed above and below the microfluidic channel (l), iii. It contains four mirrors (6) placed at an angle of 45°. It is a device according to claim 14, and its feature is that the material of the mentioned microfluidic channel (1) is silicon-based elastomer. It is a device according to claim 15, and its feature is that the said silicone-based elastomer is polydimethylsiloxane (PDMS). It is a device according to claim 14, characterized in that the wall thickness of the said microfluidic channel (1) is in the range of 200-800 µm. It is a device according to claim 17, and its feature is that the wall thickness of the mentioned microfluidic channel (1) is in the range of 400-600 µm. It is a device according to claim 14, and its feature is that the thickness of the said separator (12) is in the range of 25-200 µm. 20. It is a device according to claim 19, characterized in that the said reagent (12) is 100 µm thick. 21. It is a device according to claim 14, characterized in that the said magnet is a neodymium (N52) magnet. 22. It is a device according to claim 14, and its feature is that the microfluidic channel (1) is movable so that the said device can separate different cell types.