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Methods for screening and evaluating sperm tropism and dedicated microfluidic control system thereof

A microfluidic chip and tropism technology, applied in biochemical equipment and methods, enzymology/microbiology devices, microbial measurement/inspection, etc., can solve the problem that the impact of fluid shear force on sperm is difficult to avoid

Active Publication Date: 2013-12-04
CAPITALBIO CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantage of this device is that it is difficult to avoid the influence of fluid shear force on sperm during operation, and it can only detect a single index of chemotaxis

Method used

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  • Methods for screening and evaluating sperm tropism and dedicated microfluidic control system thereof
  • Methods for screening and evaluating sperm tropism and dedicated microfluidic control system thereof
  • Methods for screening and evaluating sperm tropism and dedicated microfluidic control system thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1, microfluidic chip

[0061] The schematic diagram of the microfluidic chip is shown in figure 1 shown. The microfluidic chip consists of four parts: a sperm inlet pool (3.2 mm in diameter), a main channel (including a 100 μm wide, 500 μm long connecting tube and a 1.5 mm wide, 1.4 mm long gradient forming tube), an outlet pool ( 3.2 mm in diameter), two branch pipes (60 degrees to the main pipe respectively), the size of the branch pipes is suitable for a field of view (830 microns x 630 microns) of the 20× objective lens of the microscope. The dotted line in the figure shows the average swimming distance of sperm from the inlet pool into the branch duct, which is about 4.5 mm. All microchannels are rectangular and 20 microns in height. Two cylindrical microstructures with a diameter of 30 micrometers are arranged at the entrances of the two branch pipes, the distance between the two cylinders is 70 micrometers, and the distance between the cylinder and ...

Embodiment 2

[0063] Example 2, microfluidic chip used in the evaluation of sperm temperature tendency

[0064] First clean the microfluidic tubing with 75% ethanol solution and deionized water respectively, treat the tubing with oxygen plasma, and fill it with 5% CO 2 , Human fallopian tube fluid (HTF) was incubated overnight at 37°C, and the inlet pool was covered with mineral oil to prevent volatilization.

[0065] Then the microfluidic chip of Example 1 is inserted as figure 2 The temperature gradient control device shown. The position where the chip is placed is the chip placement cavity 8, and the positive pressure in the outlet pool is passed into the air at a flow rate of 2 ml / hour, so that the gas-liquid interface enters the pipeline, reaching Figure 5 (c) The position shown. The part of the chip with microfluidic channels is completely surrounded by air and does not come into contact with other liquids or solids. The left and right sides of the chip placement cavity 8 are re...

Embodiment 3

[0070] Example 3, microfluidic chips are used for the capture of sperm with different motility

[0071] From the outlet pool, continue to feed air at a flow rate of 2 ml / hour to push the interface between the air and HTF towards the inlet pool. The process is as follows: Figure 5 As shown in (c)-(e), two approximately flat gas-liquid interfaces are finally formed at the entrances of the two branch pipes to prevent the exchange between the sperm in the main pipe and the sperm in the branch pipes to complete the swimming Capture of sperm by two branching ducts. The final capture effect is as follows figure 1 The magnification of the dashed area in the center is shown. Because the connecting pipe part of the main pipe is very thin, the gas-liquid interface cannot pass through the connecting pipe at a flow rate of 2 ml / hour, and a gas-liquid interface is also formed at the junction of the connecting pipe and the gradient forming pipe, such as Figure 5 (e) shown.

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Abstract

The invention discloses methods for screening and evaluating sperm tropism and a dedicated microfluidic control chip thereof. The microfluidic control chip is composed of an upper layer structure and a lower layer structure, and the surface of the upper layer structure and / or the lower layer structure includes at least one microfluid pipeline unit; each microfluid pipeline unit comprises an entrance pool, at least one main pipeline with one end communicated with the entrance pool, an exit pool communicated with the other end of the main pipeline, and at least one branch pipeline communicated with the main pipeline. While in use, a physical or chemical gradient field is formed in the main pipeline. A sample is added from the entrance pool, freely moves in the gradient field of the main pipeline, and selectively enters the branch pipelines. Through counting sperms in the branch pipelines, the sperm tropism can be quantitatively evaluated; a fluid which is insoluble or slightly soluble in a cell culture solution is introduced, an interface of the two fluids is formed at the entrance port of each branch pipeline, movement of cells between the main pipeline and the branch pipelines can be limited, so as to realize capture and recovery of the sperms having tropism.

Description

technical field [0001] The invention relates to a method for evaluating and screening sperm tropism and a special microfluidic chip system thereof. Background technique [0002] Fertilization in the human body is a complex physiological process. Many studies have confirmed that in a normal human body, sperm need to pass through layers of screening in the female reproductive tract before they can reach the fertilization site where the egg is located for fertilization. This mechanism not only ensures that the single sperm enters the egg, but also screens out healthy sperm and finally completes the fertilization. The currently recognized in vivo screening mechanisms include: sperm motility screening, only those sperm with strong motility can swim through the slender female reproductive tract to the fertilization site in the fallopian tube, and undergo acrosome reaction to complete fertilization; An important screening mechanism is the tropism screening of sperm, which mainly i...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12M1/34C12M1/38C12Q1/02C12N5/076C12N1/20
Inventor 丘天李卓琦谢兰邢婉丽程京
Owner CAPITALBIO CORP
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