In-situ Morse coding fast addressing microarray chip unit and preparation method thereof

Abstract

The invention discloses an in-situ Morse code fast addressing microarray chip unit and a preparation method thereof, the microarray chip unit comprises a super-wetting microchip array substrate and a super-wetting microchip array which is coded by referring to Morse codes, and the chip unit on a two-dimensional plane is combined with the pattern characteristics of the Morse codes. The whole chip array is divided into a plurality of areas, and the chip units in each area can be marked and positioned by using characters and numbers of Morse codes. And after the chip array substrate is subjected to ultraviolet radiation through the mask plate, forming the patterned array chip of the Morse code. The method does not depend on time-consuming and labor-consuming positioning means such as global scanning imaging and manual counting, meanwhile, the coding mode of Morse coding is combined with existing patterns in an array, the coding positioning position can also serve as a detection part, coding positioning and detection are integrated, scanning of a microscope can be efficiently and accurately guided, and the detection accuracy is improved. And the occupation of the detection area can be minimized.

Description

Technical Field

[0001] The invention relates to the field of biological detection, and in particular to an in-situ Morse coding fast addressing microarray chip unit and a preparation method thereof. Background Art

[0002] In the field of biological detection, there are two main methods for addressing specific points in an array: large-scale scanning with a microscope and para-coding positioning technology. Large-scale scanning with a microscope is a classic technical method that relies on the operator to determine the location of the point of interest through visual recognition and manual recording. This method requires high precision operation ability of the operator, especially when dealing with large-scale arrays. Accurately locating specific monitoring points consumes a lot of time and labor. In addition, the efficiency of this method is limited by the operator's manual operation speed and accuracy, and is easily affected by fatigue and human error.

[0003] In order to improve positioning efficiency and reduce human errors, the side coding positioning technology can be used. In this method, the chip to be scanned is side-coded and each point is identified with numbers or letters (such as 1, 2, 3... or A, B, C...), so as to achieve rapid positioning.

[0004] However, in actual operation, large-scale scanning with a microscope often relies on manual counting to determine the precise scanning position, which is not only time-consuming and labor-intensive, but also extremely susceptible to operator fatigue and human errors. Long periods of observation and counting may cause the operator's attention to be distracted, thus affecting the accuracy of the scanning results.

[0005] The core of the para-coding positioning technology is to determine the relative addresses of each unit of the chip by processing additional sequence marks at specific positions on the chip and tracking the sequence marks during the scanning process, thereby reducing manual intervention and improving the efficiency and accuracy of scanning. However, when integrating the coding method into the microscope array, it is necessary to consider the relationship between the chip area and the points to be tested. When the chip area is small and there are many points to be detected, the para-coding positioning system itself will occupy a part of the precious detection position. This not only limits the area that can be used for actual detection, but also may lead to partial waste of the detection area. Therefore, how to optimize the coding positioning technology so that it can guide the scanning of the microscope efficiently and accurately while minimizing the occupation of the detection area has become a technical challenge that needs to be solved urgently. Summary of the invention

[0006] Purpose of the invention: The purpose of the present invention is to provide an in-situ Morse coding fast addressing microarray chip unit and a preparation method thereof.

[0007] Technical solution: The in-situ Morse coding fast addressing microarray chip unit of the present invention comprises: a superwet microchip array substrate and a reference Morse code to encode the superwet microchip array. The chip array substrate is Morse coded in the X direction and the Y direction respectively, and the entire chip array is divided into multiple areas. Each area contains multiple sites, and each site matches its own unique code. After the chip array substrate is irradiated with ultraviolet light through a mask, a Morse coded patterned array chip is formed.

[0008] Furthermore, the entire chip adopts an integer coding method. For example, in this example, the chip is divided into 26×26 sub-unit areas, and these sub-unit areas are encoded.

[0009] Furthermore, each of the regions contains a unit site with a preset integer value.

[0010] Furthermore, the unique code of each site match is translated into a corresponding coordinate array matrix.

[0011] Furthermore, the Morse-coded patterned array chip is compatible with high-throughput imaging detection devices.

[0012] Furthermore, the Morse coded patterned array chip is suitable for fast addressing of array chips with high-density units.

[0013] The method for preparing the in-situ Morse coding fast addressing microarray chip unit of the present invention comprises the following steps:

[0014] (1) Hold the slide with tweezers and slide it back and forth at a constant speed 1 cm above the candle flame, so that a layer of candle ash is evenly deposited on the surface of the slide until the candle flame cannot be clearly seen through the slide covered with candle ash;

[0015] (2) placing the glass slide with candle ash on the surface into a sealed container, and placing 10 ml of ammonia water and 10 ml of ethyl orthosilicate in the container to cause chemical vapor deposition to deposit a layer of silicon dioxide on the candle ash layer on the surface of the glass slide;

[0016] (3) placing the glass slide after chemical vapor deposition in a muffle furnace for calcination, setting the heating rate of the muffle furnace to 10° C. per minute, heating to 550° C. and then keeping the temperature for 2 hours;

[0017] (4) The calcined glass slide was exposed to oxygen plasma for 3 minutes, and then placed in octadecylsiloxane with anhydrous toluene as the solvent and allowed to stand for 10 minutes, so that the modified glass slide was in a hydrophobic state;

[0018] (5) Placing a mask on the modified slide and irradiating it under an ultraviolet lamp, the light-transmitting part of the mask is irradiated by the ultraviolet light and becomes a hydrophilic site, while the unirradiated part remains hydrophobic;

[0019] (6) After the superwet microchip array substrate is manufactured, the Morse code of each independent secondary coding unit on the array chip is compiled to achieve addressing of the points on the superwet microchip.

[0020] Furthermore, in step (2), the reaction time is 24 hours and the reaction temperature is 37°C.

[0021] Furthermore, in step (4), the amount of anhydrous toluene used as the solvent is 50 ml.

[0022] Furthermore, the amount of octadecylsiloxane selected in step (4) is 500 μL.

[0023] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: the present invention does not rely on time-consuming and labor-intensive positioning methods such as manual counting, and at the same time combines the Morse code encoding method with the existing pattern in the array. The coding positioning part can also be used as the detection part, so that the coding positioning and detection are integrated, which can not only efficiently and accurately guide the scanning of the microscope, but also minimize the occupation of the detection area. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 is a flow chart of a method for preparing a superwet microchip array;

[0025] Figure 2 2(A) is the Morse code array diagram, 2(B) is the entire code array design diagram, 2(C) is the code array Tiles image, and 2(C) is the microscope bright field image of a single code unit and the code corresponding to each point.

[0026] Figure 3 These are bright field images of the same spot on the microarray chip observed on days 0, 1, 2, and 3. DETAILED DESCRIPTION

[0027] The technical solution of the present invention is further described below in conjunction with the accompanying drawings.

[0028] The method for preparing the in-situ Morse coding fast addressing microarray chip unit of the present invention comprises the following steps:

[0029] (1) Use tweezers to hold a glass slide and slide it back and forth at a constant speed of about 5 cm / s about 1 cm above the candle flame, so that a layer of candle ash is evenly deposited on the surface of the glass slide until the candle flame can no longer be seen through the glass slide covered with candle ash.

[0030] (2) Place the glass slide with candle ash on the surface into a sealed container. Also, place 10 ml of aqueous ammonia and 10 ml of ethyl orthosilicate in the container to cause chemical vapor deposition to deposit a layer of silicon dioxide on the candle ash layer on the surface of the glass slide. The reaction is performed for 24 hours at a temperature of 37°C.

[0031] (3) The glass slide after chemical vapor deposition was placed in a muffle furnace for calcination. The heating rate of the muffle furnace was set to 10° C. per minute. The temperature was raised to 550° C. and then kept at this temperature for 2 hours.

[0032] (4) The calcined glass slide was exposed to oxygen plasma for 3 minutes, and then placed in octadecylsiloxane (500 μL) with anhydrous toluene as solvent (50 mL) and allowed to stand for 10 minutes. The modified glass slide was in a hydrophobic state.

[0033] (5) placing a mask on the modified glass slide and irradiating it under an ultraviolet lamp, so that the light-transmitting part of the mask is irradiated by the ultraviolet light and becomes a hydrophilic site, while the unirradiated part remains hydrophobic, and thus the superwettable array chip is prepared;

[0034] (6) After the superwet microchip array substrate is manufactured, the Morse code of each independent secondary coding unit on the array chip is compiled to achieve addressing of the points on the superwet microchip.

[0035] like Figure 1 Shown is a flow chart of the preparation method of superwet microchip array units. Referring to Morse coding, we designed and prepared a microdroplet array substrate chip that can quickly locate and address functions.

[0036] like Figure 2 The figure shows the Morse code array diagram. Figure 2 (A) shows the design of the entire coding array. The substrate is Morse coded in the X and Y directions. The entire array is divided into 26×26 sub-unit areas, such as Figure 2 (B) shows the coded array Tiles image. Each sub-unit area is composed of 16 sites, so each site has its own unique code. After the substrate is irradiated with ultraviolet light through the mask, a Morse code patterned array chip is formed, as shown in the figure. Figure 2(C) shows a single coding unit microscope bright field image and the code corresponding to each point, with a scale of 200 microns. From the Morse code comparison table, we can translate the single array unit shown in 2 (C) into the corresponding coordinate array matrix. The Morse code comparison table is shown in Table 1. On the coded array chip, "·" is a circular point with a radius of 100 microns, and "-" is a square point with a side length of 200 microns in the X direction, and a rectangular point with a length of 200 microns and a width of 100 microns in the Y direction, so as to distinguish the X and Y directions.

[0037] Table 1 Morse code comparison table

[0038]

[0039] like Figure 3 As shown in the figure, the secondary unit area in the microdroplet array is (O, N), and we observe the (O0, N2) site for three days. The Morse coded microdroplet array chip provided in this experiment can address the points on the microdroplet array chip and conduct long-term regular observation of specific points of interest by simply compiling the Morse code of each independent secondary coding unit on the array chip.

Claims

1. An in-situ Morse coding fast addressing microarray chip unit, characterized in that: The invention comprises a superwet microchip array substrate and a reference Morse code for encoding the superwet microchip array. The chip units on the two-dimensional plane are combined with the pattern features of the Morse code to divide the entire chip array into multiple areas. The chip units in each area can be marked and positioned with Morse coded characters and numbers. After the chip array substrate is irradiated with ultraviolet light through a mask, a Morse coded patterned array chip is formed.

2. The in-situ Morse coding rapid addressing microarray chip unit according to claim 1, characterized in that: The entire chip array is divided into two integer secondary unit areas corresponding to the product of the X-axis and Y-axis directions.

3. The in-situ Morse coding rapid addressing microarray chip according to claim 1, characterized in that: Each of the regions contains a unit site with a preset integer value.

4. The in-situ Morse coding rapid addressing microarray chip unit according to claim 1, characterized in that: The unique code for each site match is translated into a corresponding coordinate array matrix.

5. The in-situ Morse coding rapid addressing microarray chip unit according to claim 1, characterized in that: The Morse-coded patterned array chip is compatible with high-throughput imaging detection devices.

6. The in-situ Morse coding rapid addressing microarray chip unit according to claim 1, characterized in that: The Morse-coded patterned array chip is suitable for fast addressing of array chips with high-density units.

7. The in-situ Morse coding rapid addressing microarray chip unit according to claim 1, characterized in that: Introducing asymmetric features on the XY axis of a two-dimensional plane into the coding pattern design makes it easy to identify, so the addressing and positioning of the array chip unit is independent of the placement direction of the entire chip during use.

8. The in-situ Morse coding rapid addressing microarray chip unit according to claim 1, characterized in that: The encoding method of Morse code is combined with the existing pattern in the array, and the coding location can also be used as the detection part, so that the coding location and detection are integrated.

9. A method for preparing an in-situ Morse coding fast addressing microarray chip unit, characterized in that: The steps include: (1) Hold the slide with tweezers and slide it back and forth at a constant speed 1 cm above the candle flame, so that a layer of candle ash is evenly deposited on the surface of the slide until the candle flame cannot be clearly seen through the slide covered with candle ash; (2) placing the glass slide with candle ash on the surface into a sealed container, and placing 10 ml of ammonia water and 10 ml of ethyl orthosilicate in the container to cause chemical vapor deposition to deposit a layer of silicon dioxide on the candle ash layer on the surface of the glass slide; (3) placing the glass slide after chemical vapor deposition in a muffle furnace for calcination, setting the heating rate of the muffle furnace to 10° C. per minute, heating to 550° C. and then keeping the temperature for 2 hours; (4) exposing the calcined glass slide to oxygen plasma for 3 minutes, and then placing it in octadecylsiloxane with anhydrous toluene as the solvent and letting it stand for 10 minutes, so that the modified glass slide is in a hydrophobic state; (5) placing a pre-designed through-hole mask containing a Morse code pattern on top of the modified glass slide and irradiating it under an ultraviolet lamp. The light-transmitting portion of the mask is irradiated by the ultraviolet light and becomes a hydrophilic site, while the unirradiated portion remains hydrophobic; (6) After the superwet microchip array substrate is manufactured, the Morse code of each independent secondary coding unit on the array chip is compiled to achieve addressing of the points on the superwet microchip.

10. The method for preparing the in-situ Morse coding rapid addressing microarray chip unit according to claim 9, characterized in that: The reaction time in step (2) is 24 hours and the reaction temperature is 37° C.; the amount of anhydrous toluene used as the solvent in step (4) is 50 ml; the amount of octadecylsiloxane used in step (4) is 500 μL.

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