Micro-fluidic-chip device and method for particle-and-cell sequence isolation and enumeration based on nanometer-micron channel combination

Abstract

The invention discloses a micro-fluidic-chip device and method for particle-and-cell sequence isolation and enumeration based on nanometer-micron channel combination. A micro channel of the micro-fluidic-chip device comprises a sample feeding channel, a main channel and a plurality of sample discharging channels, wherein a first electric-field isolation and enumeration channel is provided with anisolation-and-enumeration anode liquid storage hole, and is perpendicularly communicated with the main channel through a first detection door; a second electric-field isolation and enumeration channelis provided with an isolation-and-enumeration cathode liquid storage hole, and is perpendicularly communicated with the main channel and a flow field focusing channel through a second detection door;platinum electrodes are inserted into the isolation-and-enumeration anode/cathode liquid storage holes, the anode of a direct-current power source is connected with the first electric-field isolationand enumeration channel, and the cathode of the direct-current power source is connected with the second electric-field isolation and enumeration channel. According to the micro-fluidic-chip device and method for particle-and-cell sequence isolation and enumeration based on nanometer-micron channel combination, enumeration can be achieved when to-be-detected cells do not enter the detection doors, and under the condition that to-be-isolated cells are not damaged, cells accurately flow into the different discharging channels under the dielectrophoresis force.

Description

technical field [0001] The invention relates to the technical field of cell manipulation, in particular to a microfluidic chip device and method for sequentially separating and counting particles and cells based on nano-micro channel combinations. Background technique [0002] Separating different types of cells in mixed samples and accurately counting the separated cells has important significance and needs in many fields; for example, in the fields of marine environmental monitoring, biomedical research, etc., it is necessary to determine the target mixture ( There is always an urgent need for a portable device or method for rapid separation and counting of different types of cells such as bacteria, viruses, marine microorganisms, etc. [0003] There are two types of DEP separation in the field of microfluidics: direct current dielectrophoresis and alternating current dielectrophoresis. However, in DC dielectrophoresis, a voltage of hundreds of volts or even thousands of ...

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Problems solved by technology

This method uses the density and size of the particles to separate the particles, but under the physical conditions of ultracentrifugation, some cells will be destroyed, and the separable tiny particles have certain limitations.

[0009] 3) Field flow classification method: it is an important means of particle separation. The field flow classification method is to apply the field to the suspension or solution in a long and narrow tunnel, vertically (or at other angles) to The method of separating the direction of the mobile phase, this method can carry out continuous high-resolution separation of colloids with a wide size range, but the disadvantage of this method is that the sample is easy to lose, the main reason is that in the process of the interaction between the sample and the separation membrane , the separation membrane will adsorb the sample, and the pH value and ionic strength of the sample will also change.

[0010] 4) Magnetic separation method: Magnetic separation method is a method that uses magnetic force to effectively separate cells. This method requires small equipment, simple structure, and easy maintenance. However, sometimes in order to increase the magnetic field strength, a high magnetic field must be selected. There are certain technical difficulties in the selection of magnetic gathering media with saturation, and it will increase the cost of operation.

[0013] In addition, since the separation channel and the counting channel are relatively close, it is easy to interfere with each other, which affects the movement of particles and the accuracy of counting.

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