TR2022015964A2 - CLOSED CHANNEL MICROFLUIDIC PLATFORM WORKING WITH CENTRIFUGAL PRINCIPLE - Google Patents

Abstract

The invention consists of at least one microfluidic chip (3) containing at least one closed-end filling channel (2) and at least one sample chamber (1), and at least one centrifugal tube (4) designed to place this microfluidic chip (3) in the centrifuge device. It is related to biological analyzes that can be performed by using a microfluidic platform containing a microfluidic platform and liquid filling in a channel closed at one end, using the centrifuge principle on this platform.

Description

DESCRIPTION CLOSED CHANNEL MICROFLUIDICE WORKING WITH CENTRIFUGAL PRINCIPLE PLATFORM Technical Field to which the Invention Relates Filling of liquids/samples into a closed channel system using centrifugal force with the invention and manipulation was carried out. The microfluidic chip is rotated at a certain rotation speed. The liquid placed in the chamber is filled only into the closed channel that has an inlet. This principle in chemical analysis, cell analysis, diagnostic purposes and many applications in the biomedical and health fields. It has the potential to be adapted to the study.

Known Status of the Art Related to the Invention (Prior Art) Microfluidic systems work with small volumes of liquids, enabling large volumes for point-of-care testing. It has become an advantageous system. Particularly centrifugal microfluidics Systems eliminate structures such as pumps and active valves and off-chip sample processing stages, It eliminates error-prone pipetting processes (Hugo et al., 2014).

Centrifugal platforms, called lab-on-disc, are produced with low-cost fabrication. can provide processing. However, in these studies, channels and chambers were located in different layers. is positioned and has a ventilation layer to ensure fluid movement in the uppermost layer. requires (Clime et al., 2019). The microfluidics platform has a microfluidics flow in the radial direction. interaction between capillary and centrifugal force that can circulate liquid back and forth through the channel In a pumping technique based on capillary Depending on the movement, liquids can move inward along the channel (Garcia-Cordero et al., 2010). Although it is a simple and effective method, this method requires only a small amount of liquid. part of it may be displaced towards the center in a lower reservoir. Therefore, this method mixing of only a few miscible solutions or a functionalized combination of analytes Suitable for surface circulation. Two reservoirs connected via a microfluidic channel A similar mechanism occurs in the fluid flow between microfluidic disks and sudden accelerations of the It is obtained by the Euler inertia force formed by the decelerations. Euler force, microfluidics It is produced by the angular velocity of the disc in the upward direction along the channel. This force is always Since it will compete with centrifugal force, optimizations will be needed according to channel size and direction. is heard (Deng et al., 2014). In a study that pumped liquids radially inward, Under the influence of centrifugal force, a less dense sample liquid moves towards the center of rotation. It was carried out using a high-density liquid to propel (Kong et al., 2012). Sample It requires immiscible liquids or air as an interphase to maintain its integrity. is heard. Its one-way character and the need for additional fluid for pumping make this approach It is limiting. In a passive pumping system presented, the liquid is in the branch in an adjacent channel. There is a principle based on the pneumatic compression of air with the hydrostatic pressure produced. is taking. In this study, the accumulated pneumatic energy was reduced by reducing the rotation speed of the platform. is released and used to pump the fluid back to the return center (Clime et al., 2019).

Although the method is sensitive and reproducible, it requires the production of additional compression chambers. hears. In a centrifugal system using a siphon valve, the siphon structure moves the liquids at a rotation speed. It can either discharge within the range or cut off the flow by increasing the return frequency (Zhu et al., 2018). Discontinuous The siphon valve consists of an additional air hole at the top of the siphon. When rotation is stopped The siphon is fed, and when the disc starts to rotate, if the rotation speed is high enough, the liquid in the siphon moves away from the center. Thus, air enters the siphon through the air hole at the top of the siphon. It will cut off the fluid in the channel. At lower rotation speed, the gas-liquid near the air hole It forms a meniscus at the interface. The liquid flows from the loading chamber to the collection chamber via the siphon. is transmitted. The design and production of these centrifugal systems containing siphon structures is complex. As in this study, the disk is exposed to constant external forces while rotating, and the gap between the liquid and gas The interface will depend on stabilization at specific locations. Hydrophilic and hydrophobic in a centrifugable disk that can separate serum from blood with a system compatible with biomarkers, cross-flow filtration method separates serum and retains amphiphilic biomarkers in serum (Lenz et al., 2021). The device consists of chambers located on top of each other, separated by a membrane. and there are four separation units with different functions on the disk. Cross-flow filtration During the process, the sample passes tangentially through the filter by centrifugal force. membrane Components smaller than the pores pass through the filter as the pressure increases, while larger components remain on the membrane surface. Hematocrit in a closed channel on a polymer disc In a study that provides measurement, an inlet chamber on the disc, an overflow channel with a hydrophobic valve and A two-layer capillary channel was created (Riegger, Lutz et al., 2007). Centrifugal assisted capillary filling, two-layer channel with different levels (upper level wider than lower-middle level) It is achieved by the use of ducts and the special shaping of the closed channel end. capillary force It carries the blood to the closed capillary channel, supported by a centrifuge. When the channel reaches the capillary end, it is reversed. Filling in the direction begins, and with the opening of the hydrophobic valve, excess blood fills the chamber here and Hematocrit value is measured by sedimentation of red blood cells. This method is just fine The manufacturing process requires microfluidic chips with a special calculated channel cross-sectional area. It is laborious and costly. Liquid filling into the channels is done using the air permeability of PDMS. In this study, density-gradient solution and microparticles were introduced into the channels under vacuum. (Oksuz & Tekin, 2021). Entrance for centrifugation of the microfluidic system Separation of microparticles of different densities by preventing blockage with this system has been achieved. Although the system is an advantageous study for cell separation, different liquid It is not a system suitable for manipulations, and it is not possible to place samples in channels under vacuum. It also takes quite a long time to load.

Active elements; vacuum, magnetic, electrical and mechanical forces for fluid flow and control It requires the application of external force such as Liquid induced by centrifugal force The unidirectional character of their flow is a fundamental limitation in the design of microfluidic circuits. It leads. In a study using an active pumping element, permanent magnets were used. It includes a pump system that is sealed with deformable polymer layers with which it is integrated. (Haeberle & Zengerle, 2007). Rotation of rooms in a constant magnetic field By ensuring that the air is compressed in deformable rooms, a constant fluid flow begins. Active Centrifugal microfluidic systems using elements offer interesting methods. Sets that need to be mounted on a rotating platform for their application and their requires checking. Installation of electronic pumps and electromechanical In a centrifuge platform made by using valves, while the platform rotates at high speed Creates air pressure at pressure ports on the chip via a pneumatic connection (Clime et al., 2019). The resulting air pressure interacts with the circuit elements on the chip. It performs functions such as valving, reverse pumping, and bubble mixing. But like this The cost required to produce a complex platform is increasing.

At stages such as diagnosing many diseases in the clinic and monitoring the disease process, The sample taken from the patient must be pre-treated and centrifuged.

Microfluidic systems feature centrifuge, which is an indispensable step in the clinic. can be demonstrated with centrifugal microfluidic systems. As an on-disk laboratory Passive pumping methods in the so-called lab-on-a-disc systems are examined. time fluid movement chambers, ventilation holes, siphons located in different layers It can be provided with structures, membranes or valves. These designs both increase the cost In addition to restricting fluid movement, it cannot guarantee efficient fluid transfer.

Additionally, since the elements used for liquid manipulation are sensitive, the method should be well optimized. It requires additional compression or waste chambers to prevent the liquids from overflowing.

Each chamber and layer added to microfluidic systems increases the cost and ease of use. directly affects. When looking at active pumping systems, externally mechanical, Studies using electrical or magnetic forces involve many complex Although there are methods that allow manipulation and analysis, they are based on a rotating platform. The components that need to be installed limit the usability of the system. Also previous Considering the techniques, most centrifugal microfluidic systems are disk-shaped. is designed. This is done in the form of a disk reader in order to increase the rotation speed of the designed system. This means that a rotating platform, i.e. an additional device design, will be needed. In summary, the technical problem encountered in the prior art is the solution for liquid manipulation and sample analysis. centrifugable microfluidic systems are expensive, complicate production, and The use of structures and methods that cannot guarantee Additional device for people who will use it separately It also needs to be taken.

Brief Description and Purposes of the Invention The present invention meets the above-mentioned requirements and eliminates the disadvantages. and a microfluidic platform that brings some additional advantages.

In the invention, the filling method presented by simplifying the production and all the materials used in the art are used. Additional structures were eliminated. In this way, it is compatible with the centrifuge device found in every laboratory. has been made. What makes the filling principle unique is the different characteristics of each channel (width, hydraulic resistance changes according to height, length) and accordingly the need for filling The rotation speed (RPM) heard also changes. As hydraulic resistance increases, the required rotation speed increases. The pressure in the channel with the applied centrifugal force is increased and air is released into the sample chamber through the channel, filling the channel. is provided.

Centrifugal microfluidic systems are used in the laboratory on disk or on CD in technology. It is known as the laboratory and is used to manipulate liquids by applying centrifugal force. systems that provide. In classical microfluidic systems known in the literature, channel filling is the entrance. and is provided with exit holes. A closed channel for applying centrifugal force to the channel. should be used and this can be done with centrifugal microfluidic systems known in the art. For this to happen, active and passive valves, membranes and ventilation holes must be used.

In the invention, for the first time, a flat closed system was created without the use of any additional structure (valve, siphon, membrane, etc.). Fluid manipulation was performed within the channel. Additionally, disc and CD products offered in the art An additional device for applying centrifugal force in laboratory systems is required (like a CD rom or designed in this way). The method and design presented with the invention Since it is suitable for centrifugal devices, it does not require such devices. In this way The application area is also expanding.

The primary purpose of the invention is to provide structures such as additional chambers, valves, membranes and siphons that provide filling. Closed channel filling and analysis in a single channel with only centrifugal force without the need for To develop a centrifugal microfluidic system that provides In the invention, liquid was poured into a chamber. placement and filling into the channel and carrying out the necessary operations are completely done by centrifugal force. It is carried out with .

With the invention filling method, all additional structures used in the technique were eliminated. In this way, every It was made compatible with the centrifuge device in the laboratory. What makes the filling principle unique is the hydraulic resistance of each channel according to its different characteristics (width, height, length). change and accordingly the rotation speed (RPM) needed for filling is to change. As the hydraulic resistance increases, the required rotation speed increases. Applied With the centrifugal force, the pressure in the channel is increased and the sample is transferred to the sample chamber through the channel. Filling is achieved by providing air outlet.

The invention microfluidic system is suitable for many biological analysis processes (blood sample analysis, DNA analysis, purification, separation, enrichment, single cell) and most of them are compatible with microscopy. It is a system that allows it to be viewed as well as analyzed by phone.

The invention solved the main problems in previous techniques: cost, complex design and an additional rotation. In addition to providing an advantage by eliminating the need for a device, additional compression chambers, without using siphon and membrane structures, using a single inlet chamber in two directions (inlet and exit) and offers a principle not included in the literature. Presented Thanks to this method, the centrifuge device in the research, cell and diagnostic laboratory has low volume (capable of processing samples, providing short-term analysis, sample pretreatment) A method that has the potential to eliminate it is aimed. desired method and According to the analysis, the channel design was developed. All you need to get results is is the placement of the microfluidic chip in the centrifuge device. In this way, other methods When compared, the invention provides a great advantage in terms of cost and ease of use. It will maintain this advantage in terms of The method also has a programmable structure. It allows many analysis steps to be carried out automatically in the centrifuge device. It provides.

With the invention product and the method used, it is the first time that a closed channel can be placed in a closed channel without using any additional structure. filling is provided. What makes the filling principle unique is the different characteristics of each channel (width, hydraulic resistance changes according to height, length) and accordingly the need for filling The rotation speed (RPM) heard also changes. As hydraulic resistance increases, the required rotation speed increases. The pressure in the channel with the applied centrifugal force is increased and filling is ensured by allowing air to exit through the channel into the sample chamber. What makes the product unique is that, depending on the filling principle, it is liquid compatible with centrifuge devices. It has become a product that enables manipulation and sample analysis.

Centrifugal microfluidic systems are used in the laboratory on disk or on CD in technology. It is known in the art as a laboratory and by applying centrifugal force, liquid systems that enable manipulation. In classical microfluidic systems known in the literature Channel filling is provided by inlet and outlet holes. Applying centrifugal force to the channel A closed channel must be used and centrifugal microfluidic systems known in the art To achieve this, active and passive governors, membranes, ventilation holes, special channels are used. architectures should be used. In this invention, for the first time, any additional structure (valve, siphon, membrane, etc.) Liquid manipulation was carried out in a straight closed channel without the use of Moreover Application of centrifugal force in laboratory systems on disk and CD offered in the technique requires an additional device (such as a CD-ROM or designed to do so). presented in the invention Since the method and design are suitable for the centrifuge device commonly found in the laboratory, this type of It does not require any devices. In this way, the application area also expands. The chip can be filled from different reservoirs sequentially. Different channel sizes and centrifuge Using the speeds, liquid can only be drawn into the chip from the desired reservoir. Thus For molecular analysis, solutions can be introduced sequentially into the chip. By adjusting the channel size The desired volume of liquid can be drawn from the reservoir. Thus, a precise amount of liquid is available for analysis. can be used. With the invention, hematocrit and white blood cell amount can be determined and Plasma can be collected on the chip.

Definitions of Drawings Explaining the Invention Figure 1: Closed channel microfluidic design (A) Microfluidic chip parts (top view) (B) Microfluidic chip created using double-sided tape, a closed-end filling channel and Contains sample chamber (side view).

Figure 2: Centrifuge tube designed to place the microfluidic chip in the centrifuge device Figure 3: Filling rate of the microfluidic chip depending on channel width and channel height change (A) At the same time, channels with different channel widths and channel heights Rotation speed required to fill (B) Hydraulic resistance and rotation speed depending on the channel height (C) Relationship between hydraulic resistance and rotation speed depending on channel width Figure 4: At a fixed channel width and height, the channel changes over time depending on the channel length. connected filling profile Figure 5: By keeping the channel width and height constant, hydraulic resistance and relationship of filling speed Figure 6: Incubation of the whole blood sample in the microfluidic chip for 5, 10 and 15 minutes, respectively Buffy coat and plasma regions formed as a result of centrifugation Figure 7: (A) Calculated hematocrit value at different rotation speeds (B) Plasma after centrifugation Figure 8: Comparison of hematocrit values measured on the chip with a microhematocrit tube comparison with measurements Figure 9: Correlation between buffy coat thickness and white blood cell count Definitions of the Elements/Parts/Parts Constituting the Invention Prepared to better explain the microfluidic platform developed with this invention. The parts/sections/elements in the figures are stated below. 1: Sample chamber 2: Filling channel 3: Microfluidics Chip 4: Centrifuge Tube Detailed Description of the Invention The invention is a microfluidic platform for liquid filling and its features are; ° At least one filling channel (2) with one end closed and flat cross-sectional area and at least one filling channel connected to the channel (2). microfluidic chip (3) containing the sample chamber (l) and ° At least one centrifuge designed to place the microfluidic chip (3) into the centrifuge device It contains the tube (4).

Filling channel (2) according to channel length (h), channel width (W) and channel height (L) The rotation speed of the centrifuge device in which the microfluidic chip (3) is positioned can be adjusted or It is characterized by being programmable.

The microfluidic chip (3) included in the invention has a length of 5-50 mm and different widths (0.1-5 mm). It consists of a flat and closed filling channel (2) (Figure 1 (A)). Microfluidics chip (3) 0.15 mm thickness double-sided tape, the lower surface of the coverslip with a thickness of 1 mm, the upper surface of the sample It contains a Polymethyl methacrylate (PMMA) layer containing a chamber (l) and a closed filling channel. (Figure 1 (B)). The volume of the sample chamber (l) is 1-100 uL.

The liquid filling method includes the following process steps: ° At least one flat cross-sectional filling channel (2) with one end closed and at least one channel connected to the channel (2). containing the sample chamber (l), filling channel (2) channel length (h), channel width (W) and channel At least one microfluidic chip is used to adjust the rotation speed of the centrifuge device according to its height (L). (3) ensuring, ° At least one centrifuge designed to place the microfluidic chip (3) into the centrifuge device ensuring the tube (4), ° Filling the liquid into at least one sample chamber (l), ° Placing the microfluidic chip (3) into the centrifuge tube (4), ° Filling channel (2) according to channel length (h), channel width (W) and channel height (L) Adjusting the centrifuge device rotation speed, ° Centrifuge the microfluidic chip (3) by rotating it for a certain period of time at the set centrifuge speed. determined liquid by allowing air to exit from the channel (2) to the sample chamber (1) with the force filling the volumes from the sample chamber (1) into the channel (2).

The invention method is performed by rotating the microfluidic chip (3) at different centrifuge speeds for specified periods of time. Programmable flow of different liquids from the desired sample chamber (l) to the filling channel (2). It is used for filling.

After the microfluidics chip (3) is placed in the liquid reservoir to be filled in the channel (2), a special It was put into the centrifuge tube (4) designed as a centrifuge and placed in the centrifuge device (Figure 2). The centrifuge tube (4) fits perfectly inside the microfluidic chip (3) and is placed in the centrifuge device. It is a tube that ensures that the chip (3) remains stable. Technically fits perfectly into the centrifuge device, inside which the microfluidic chip (3) can enter and which prevents the movement of the chip (3) during centrifugation. contains at least one indent.

The microfluidic chip (3) is rotated at different centrifugal speeds (100-1000 rpm) to produce different liquids. volumes can be filled into the channel (2). Each channel (2) features (length, width, It has different hydraulic resistance due to height). The increase in hydraulic resistance causes the filling of the channel (2) to decrease. This means that it will need a higher rotation speed. For this reason, every channel The aim here is to find the rotation speed at which the channel (2) is filled according to its characteristics. Different channel (2) sizes can be filled at different rotation speeds (Figure 3A). This is on the chip (3) If different channel (2) widths connected to the chamber are used, different rotation speeds can be programmed It means that it will create a flow profile. In this way, different liquids are Processes that require mixing can be carried out in the centrifuge device with the recommended principle.

In this invention, filling of the closed channel (2) system consists of valve, membrane and siphon structures. It is presented for the first time without being used.

With the invention, different channel (2) sizes can be filled at different rotation speeds at the same time. is observed (Figure 3A). According to the results obtained, channel (2) width and channel height It has been concluded that as it increases, the rotation speed required for the liquid to fill into the channel (2) decreases. The filling principle was revealed by calculating the hydraulic resistance with Equation 1 (Equation 1).

Hydraulic resistance (R), channel height (11), channel width (W), channel length (L) and liquid It is related to its viscosity (u). Hydraulic resistance depending on channel width and channel height When the relationship between the rotation speed required for filling and the calculation is taken into account, the correlation can be seen (Figure 3B, C). This means different channels (2) connected to reservoirs on the chip (3). If widths are used, it will create a programmable flow profile at different rotation speeds. It means. In this invention, filling of the closed channel (2) system consists of valve, membrane, It is presented for the first time without the use of siphon structures. 1-0.63(g)h3W The channel width of the channels is kept constant at 2 mm and the channel height is 150 um, depending on the channel length. According to the time-dependent volume profile was examined (Figure 4). Hydraulic resistance using equation 1 The relationship between the channel length and the rotation speed required to fill the channel is calculated. is shown (Figure 5). Thus, the channel with low hydraulic resistance, that is, the channel length is short, While it takes a shorter time for it to reach 95% of its volume, as the channel length increases, this duration is also included.

Within the scope of the invention, hematocrit and white blood tests performed routinely in the hematology unit Cell counting was applied in a closed channel (2) system. 10 ML whole blood sample sample placed in the chamber (l) and then in the centrifuge device at different rotation speeds and times. centrifuged. In this way, plasma is obtained from whole blood without using any additional processes or additional structures. has left. The total number of red blood cells collected in the lower part of the channel is within the channel (2). The hematocrit value can be calculated from its ratio to the sample volume (Figure 8). The result obtained Considering that the effects of rotation speeds on hematocrit calculation are the same. Optimum rotation speed and duration by counting the amount of cells remaining in the plasma is 4000 rpm 10 It is determined as minutes (Figure 7A, B). In the clinic, it is used in blood for plasma separation. All cells present must collapse and there should be no cells left in the plasma. is required. For this reason, the number of cells remaining in the plasma at rotation speeds It was observed that there was no difference in cell numbers between the two.

The separation of plasma from the whole blood sample and the remaining white blood between the red blood cells It was observed that the buffy coat region containing cell and platelet was obtained in the invention (Figure 6). buffy The amount of white blood cells in the blood can be calculated by looking at the coat thickness (Figure With the presented technique, for the first time, a closed-ended channel was used to create additional structures with centrifugal force from a single chamber. hematocrit value and The amount of white blood cells could be determined. This discovery was based on various blood tests separation by serving applications at different molecular levels as well as being applicable It also has the potential to be used in studies such as health and enrichment.

Considering the filling principle, in the direction of rotation of the chip, the liquid moves towards the channel surface. It is seen that it forms a meniscus structure. The meniscus structure advances along the channel and absorbs the fluid. It fills starting from the bottom of the duct, and the air in the duct is discharged in the opposite direction. is done.

By making changes in channel shapes and positions on the designed system, water can be removed from different chambers. sequential sample filling and mixing of different samples in the channel can be provided. Additionally, with the designed channel and the proposed principle, plasma from whole blood can be obtained. separation of biomarkers (such as creatinine) in the separated plasma can be achieved. measurement can also be made. For this, the whole blood sample is placed in the sample chamber and It is then centrifuged. In this way, the red blood cells in whole blood are at the bottom. While collecting, there is an area in the middle where white blood cells, known as buffy coat, collect. is formed and pure plasma remains on top. Red blood from microfluidics chip image The area covered by cells is measured and compared to the entire sample area, and the hematocrit value is determined. will be found. In addition, white blood cells collect in the area called buffy coat in the middle. The number of white blood cells can be calculated from this region. in plasma For the measurement of biomarkers, the blood sample is used as a result of its reaction with biomarkers outside the chip. It is preferably mixed with solutions that yield colorimetric products and given to the sample chamber.

Then, the mixture is transferred to a closed channel by centrifugation and the red blood is collected by high-speed centrifugation. cells and white blood cells are precipitated. In the plasma region above biomarkers react with the solutions, depending on the concentration of the biomarker found. Depending on the color channel, it produces color at different intensities. According to this color intensity biomarker concentration can be calculated. With this method, blood in the same channel By separating blood cells from plasma, the negative effect of blood cells on colorimetric measurements is eliminated. (such as background noise) are eliminated. The proposed invention is used in cell separation, single cell It can be used in studies and many molecular and clinical tests that require centrifugation. has potential. In addition, the proposed system can process a very small amount of samples (0,, The separated samples can also be examined under a microscope due to the transparent structure of the microfluidic chip. can be examined. The separated samples were collected from the microfluidic chip and collected into different samples. It can also be used in analyses.

Claims (14)

STEMLER is a microfluidic platform and its features are; ° Microfluidic chip (3) containing at least one filling channel (2), closed at one end and with a flat cross-sectional area, and at least one sample chamber (l) connected to the channel (2), ° Designed to place the microfluidic chip (3) in the centrifuge device, at least It is characterized by containing a centrifuge tube (4) and the rotation speed of the centrifuge device in which the microfluidic chip (3) is positioned according to the filling channel (2), channel length (h), channel width (W) and channel height (L) being adjustable or programmable. It is a microfluidic platform in accordance with claim 1 and its features are; The length of the filling channel (2) with one end closed is 5-50 mm. It is a microfluidic platform in accordance with claim 1 and its features are; The width of the said one-end closed filling channel (2) is 0.1-5 mm. It is a microfluidic platform in accordance with claim 1 and its features are; The sample chamber (l) consists of a material layer selected from the group containing glass, polystyrene, polyVinyl chloride, cyclic olefin co-polymers, polycarbonate, polydimethylsiloxane (PDMS)) and Polymethyl methacrylate (PMMA). It is a microfluidic platform in accordance with claim 1 and its features are; The sample chamber (l) has a volume of 1-100 uL. It is a microfluidic platform in accordance with claim 1 and its features are; The centrifuge tube (4) is such that the microfluidic chip (3) can enter and contains at least one recess that prevents the movement of the chip (3) during centrifugation. It is a microfluidic platform suitable for the claim and its features are; It is used in liquid filling. It is a microfluidic platform in accordance with claim 1 and its features are; It is used in hematocrit analysis. It is a microfluidic platform suitable for the claim and its features are; It is used to determine the amount of white blood cells. It is a microfluidic platform in accordance with claim 1 and its features are; It is used for plasma separation and taking the plasma out of the chip. 11. It is a microfluidic platform in accordance with claim and its feature is; It is used in biomarker analysis. 12. It is a microfluidic platform according to claim 1 and its feature is; It is used to examine the sample inside the chip under a microscope. 13. It is a liquid filling method and its feature is; a) The filling channel (2), which is closed at one end and contains at least one filling channel (2) with a flat cross-sectional area and at least one sample chamber (1) connected to the channel (2), has the channel length (h), channel width (W) and Providing at least one microfluidic chip (3) in which the rotation speed of the centrifuge device is adjusted according to the channel height (L), b) Providing at least one centrifugal tube (4) designed to place the microfluidic chip (3) into the centrifuge device, c) Providing the liquid at least filling a sample chamber (1), d) placing the microfluidic chip (3) into the centrifuge tube (4), e) filling channel (2) centrifugation according to channel length (h), channel width (W) and channel height (L). adjusting the rotation speed of the device, f) The process of filling the determined liquid volumes from the sample chamber (1) into the channel (2) by rotating the microfluidic chip (3) at the set centrifuge speed for a certain period of time and allowing air to exit from the channel (2) to the sample chamber (1) with the centrifugal force. It contains the steps. 14. It is a method in accordance with claim 13, and is used to programmably fill different liquids from the desired sample chamber (1) into the filling channel (2) by rotating the microfluidic chip (3) at different centrifuge speeds for specified periods of time.