A virgin fly collecting device and method for fruit fly genetic research

Abstract

The present application relates to the technical field of insect screening and collecting, in particular to a virgin fly collecting device and method for fruit fly genetic research; the device comprises a guide pipe and a camera assembly and the like structure, two elastic bags are arranged on the guide pipe, and a preset interval is arranged between the two elastic bags as a detection section; the camera assembly is arranged opposite to the detection section, and is used for shooting photos of fruit flies located in the detection section; real-time image acquisition of fruit flies passing through the detection section is realized through the camera assembly; virgin flies and non-virgin flies are automatically distinguished by combining a deep learning visual recognition program built in an upper computer; and the airflow direction is controlled by an electromagnetic three-way valve to drive the elastic bag and the air blowing pipe to complete the shunting of fruit flies; the whole process does not need manual anesthesia, magnifying glass identification and the like operation, the labor cost is significantly reduced, and the problems of long screening time and large labor intensity of virgin flies in the prior art are solved.

Description

Technical Field

[0001] This invention relates to the field of insect screening and collection technology, and in particular to a device and method for collecting virgin flies for fruit fly genetic research. Background Technology

[0002] The fruit fly (Drosophila melanogaster), belonging to the family Drosophila in the order Diptera, is a widely used model organism in genetics, developmental biology, neuroscience, and medical research. Fruit flies are characterized by rapid reproduction, short generation cycles, a small chromosome number (2n=8), and high gene conservation. Studies have shown that approximately 75% of human disease-related genes have homologous genes in fruit flies. Fruit flies are easily reared in large numbers in the laboratory using culture flasks and artificial culture media, and a rich resource of mutants has been established, making them an indispensable core experimental material in life science research.

[0003] In genetic research, fruit flies need to be cultured. Wide-mouthed bottles or large test tubes are typically used, filled with solid culture medium and sealed with breathable sponges or cotton balls wrapped in gauze. After maturing, the flies mate and lay eggs. The eggs are then transferred to new culture bottles to await pupation. Unmated female fruit flies that have pupated are called virgin flies and are crucial in genetic research. Therefore, identifying virgin flies is critical. Within 8–10 hours of emergence, the female flies' reproductive organs are not fully developed, so they do not mate. This is the time to collect virgin flies. The current method for collecting fruit flies during their window period involves anesthetizing newly emerged fruit flies with anesthesia bottles or anesthetic plates, then pouring them into a petri dish and immediately identifying them one by one using a magnifying glass. Virgin female flies are large, with slender abdomens that are pointed at the end, and five black rings on their abdomens. They lack sex combs on their forelegs and have simple external genitalia. After mating, the abdomen of female flies is noticeably swollen and the end becomes rounded. Male flies are small, with short, round abdomens that are blunt at the end. They have three rings on their abdomens, the last of which is thick and connected. They have black sex combs on the first tarsal segment of their forelegs and complex external genitalia.

[0004] The process of separating virgin fruit flies is very time-consuming and tedious. To allow researchers more time for experimental research, designing a device that can automatically screen and collect virgin fruit flies is crucial. Currently, there are many designs available, such as a Chinese patent application (application number 201510399902.1) that discloses a male-female fruit fly separator. This separator uses a Y-shaped tube and a manually operated switch to screen and collect male and female fruit flies. However, its level of automation is low, still requiring manual operation. Currently, there are also vision-based solutions for screening male and female fruit flies. These involve placing newly emerged fruit fly pupae in a test tube. When the flies emerge, they are lured through a narrow channel by a scent from the other end of the tube. A camera below captures microscopic images of the fruit flies' abdominal spots, reproductive organs, etc., which are then transmitted to a computer for rapid analysis and activation of the screening mechanism. The virgin flies are then automatically collected. This technology is still under research.

[0005] To address this, the inventors have proposed a collection device and method for automating the screening and collection of virgin flies. Summary of the Invention

[0006] The purpose of this invention is to provide a device and method for collecting virgin flies for Drosophila genetic research, thereby solving the problems mentioned in the background art. The specific technical solution is as follows:

[0007] The first objective of this invention is to provide a virgin fly collection device for fruit fly genetic research, comprising:

[0008] The guide tube has two elastic bladders with a preset interval between them as a detection section. The detection section is transparent and the elastic bladders expand in their natural state, compressing the inner diameter of the guide tube.

[0009] A camera assembly is arranged opposite to the detection section and is used to take pictures of fruit flies located in the detection section. A host computer is electrically connected to the camera assembly and is used to perform image recognition on the fruit fly pictures taken by the camera assembly to determine whether they are virgin flies.

[0010] An air blowing tube and a separation tube are respectively installed on both sides of the detection section, and a one-way valve is installed at the proximal end of the separation tube;

[0011] An electromagnetic three-way valve has an inlet connected to an air supply module, a first exhaust port connected to the air blowing pipe, and a second exhaust port connected to a negative pressure generator via a pipeline. The negative pressure interface of the negative pressure generator is connected to the two elastic bladders via a negative pressure pipeline. The electromagnetic three-way valve is connected to the host computer and is used to control the airflow through the negative pressure generator to cause the elastic bladders to contract, or to switch the airflow to the air blowing pipe to blow non-virgin flies in the detection section into the separation pipe.

[0012] Preferably, the guide tube has a first connector at its proximal end and a second connector at its distal end. The first connector is used to connect to an incubation bottle, the second connector is used to connect to a virgin fly collection bottle, and the separation tube has a third connector at its distal end for connecting to a first fruit fly collection bottle.

[0013] Preferably, a first outer sleeve is fitted over the guide tube, forming a male fruit fly passage between the first outer sleeve and the guide tube; a plurality of small holes are provided on a section of the guide tube inside the first outer sleeve, the small holes allowing only male fruit flies to pass through; the first outer sleeve is provided with a connecting tube, the end of which is connected to a second fruit fly collection bottle.

[0014] Preferably, a plastic film is provided outside the small hole, and the plastic film forms a one-way channel to prevent fruit flies from returning from the first outer tube to the guide tube.

[0015] Preferably, the small hole has an elliptical structure with a major axis of 0.8-1.0 mm and a minor axis of 0.6-0.8 mm.

[0016] Preferably, the inner wall of the first outer tube is coated with a pheromone coating to attract male fruit flies to enter the first outer tube through the small hole.

[0017] Preferably, a second outer sleeve is also fitted onto the separating tube, and a male fruit fly channel is provided between the second outer sleeve and the separating tube; the separating tube has sieve holes for connecting the second outer sleeve.

[0018] Preferably, the inner diameter of the guide tube is 1.5-2.0 mm.

[0019] Preferably, the camera assembly further includes an infrared sensor, which automatically triggers the camera assembly to take pictures when the infrared sensor detects a fruit fly entering the detection section.

[0020] A second object of the present invention is to provide a method for collecting virgin flies using any of the above-described devices, comprising the following steps:

[0021] S1: Install the first connector to the incubation bottle, the second connector to the virgin fly collection bottle, and the third connector to the first fruit fly collection bottle. Place food to attract fruit flies in the virgin fly collection bottle, and arrange the guide tube upward or tilted upward.

[0022] S2: Open the air supply module, and the airflow enters the negative pressure generator through the electromagnetic three-way valve, causing the elastic bladder to contract, and the fruit fly passes through the detection section;

[0023] S3: The camera component captures images of fruit flies entering the detection section and sends them to the host computer, which then performs image recognition.

[0024] S4: When the identification result is a virgin fly, maintain the current state of the electromagnetic three-way valve, and the fruit fly continues to move forward to the virgin fly collection bottle;

[0025] S5: When the identification result is non-virgin flies, the host computer controls the electromagnetic three-way valve to switch the airflow direction, the negative pressure of the negative pressure generator disappears, the elastic bladder resets to fix the fruit flies on the front and back sides of the detection section, and the airflow blows the non-virgin flies in the detection section into the separation tube through the blowing pipe, and finally into the first fruit fly collection bottle.

[0026] S6: After the blow-off is completed, the electromagnetic three-way valve is reset, the airflow re-enters the negative pressure generator, and the fruit fly resumes crawling.

[0027] Beneficial effects:

[0028] 1. This invention uses a camera component to collect real-time images of fruit flies passing through the detection section. Combined with a deep learning visual recognition program built into the host computer, it automatically distinguishes between virgin and non-virgin flies. An electromagnetic three-way valve controls the airflow direction to drive the elastic bladder and the blowing tube to complete the separation of fruit flies. The entire process does not require manual anesthesia, magnifying glass identification, or other operations, which significantly reduces labor costs and solves the problems of long time consumption and high labor intensity in the screening process of virgin flies in the prior art.

[0029] 2. Two elastic bladders are installed inside the guide tube. The contraction and repositioning of the elastic bladders are controlled by a negative pressure generator. When the elastic bladders contract, fruit flies can pass smoothly through the detection section. When they reposition, they compress and fix the fruit flies on both sides of the detection section. This ensures that each fruit fly enters the detection section one by one for testing, and effectively prevents misjudgment and missed detection caused by multiple fruit flies passing through at the same time. The diameter of the guide tube is limited to 1.5-2.0mm to prevent fruit flies from passing through side by side, fundamentally preventing fruit flies from mating inside the guide tube.

[0030] 3. The first outer sleeve, fitted onto the outside of the tube, attracts male fruit flies through a small hole into the second fruit fly collection bottle via a pheromone coating. This pre-screens out a large number of male fruit flies before they enter the detection section, significantly reducing the number of flies that need to enter the visual recognition stage and greatly improving overall screening efficiency. The second outer sleeve on the separation tube, with appropriately enlarged sieve apertures, can further recover male fruit flies blown into the separation tube, reducing waste of male fruit flies. Attached Figure Description

[0031] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0032] Figure 1 This is a schematic diagram of the structure of the device described in this invention;

[0033] Figure 2 This is a cross-sectional view of the guide tube described in this invention. Detailed Implementation

[0034] The following detailed description, in conjunction with the accompanying drawings and specific embodiments, provides a further detailed explanation of the virgin fly collection device for Drosophila genetic research proposed in this invention. The advantages and features of the invention will become clearer from the following description. It should be noted that the drawings are all in a very simplified form and use non-precise proportions, and are only used to facilitate and clarify the illustration of the embodiments of the invention.

[0035] In the description of this invention, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this invention, "a plurality of" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0036] It should be noted that the illustrations provided in the following embodiments are only schematic representations of the basic concept of the present invention. Therefore, the illustrations only show the components related to the present invention and are not drawn according to the actual number, shape and size of the components in the actual implementation. In the actual implementation, the state, quantity and proportion of each component can be arbitrarily changed, and the layout of the components may also be more complex.

[0037] In some embodiments, a virgin fly collection device for fruit fly genetic research, such as Figure 1-2As shown, the device includes a guide tube 1, with a first connector 11 at its proximal end and a second connector 12 at its distal end. The first connector 11 is connected to an incubation bottle, and the second connector 12 is connected to a virgin fly collection bottle. The guide tube 1 has two elastic bladders 13, with a spacing of 3.6-4.0 mm between them, sufficient to accommodate one fruit fly. The elastic bladders 13 expand naturally, compressing the inner diameter of the guide tube 1 to compress and fix the fruit fly. The guide tube 1 located between the two elastic bladders 13 is a detection section 14. The detection section 14 is transparent, or the entire guide tube 1 is made of transparent material. A camera assembly 2 is located below the detection section 14, connected to a host computer. The host computer has a built-in visual recognition program that performs image recognition on the fruit fly photos taken by the camera assembly 2 to determine whether the fly is a virgin. The detection section 14 has two sides... The device is equipped with an air blowing pipe 15 and a separation pipe 16. A one-way valve 161 is installed at the proximal end of the separation pipe 16. After air is blown into the air blowing pipe 15, the air pressure will open the one-way valve 161, allowing the fruit flies located in the detection section 14 to be blown into the separation pipe 16 by the airflow. A third connector 162 is installed at the distal end of the separation pipe 16. The third connector 162 is connected to the first fruit fly collection bottle. The air blowing pipe 15 is connected to the first exhaust port of the electromagnetic three-way valve 3. The air inlet of the electromagnetic three-way valve 3 is connected to the air supply module 4. The second exhaust port of the electromagnetic three-way valve 3 is connected to the negative pressure generator 5 through a pipeline. The electromagnetic three-way valve 3 is used to control the airflow through the negative pressure generator 5 or through the air blowing pipe 15. The negative pressure interface of the negative pressure generator 5 is connected to two elastic bladders 13 through a negative pressure pipeline. When the airflow from the air supply module 4 passes through the negative pressure generator 5, the air pressure in the elastic bladder 13 decreases, the elastic bladder 13 contracts, and the fruit flies can pass through smoothly.

[0038] In use, the first connector 11 is installed on the incubation bottle, the second connector 12 is installed on the virgin fly collection bottle, and the third connector 162 is connected to the first fruit fly collection bottle. Food that attracts fruit flies is placed inside the virgin fly collection bottle. The guide tube 1 is positioned upwards or tilted upwards, utilizing the scent of the food and the fruit flies' tendency to climb, causing them to crawl into the guide tube 1. To prevent mating, the diameter of the guide tube 1 is set to 1.5-2.0 mm to avoid fruit flies running side-by-side and thus preventing mating within the guide tube 1. After the fruit flies crawl into the guide tube 1, the air supply module 4 is opened, and the airflow enters the electromagnetic three-way valve 3 and then the negative pressure generator 5. The elastic bladder 13 contracts, allowing the fruit flies to pass normally through the detection section 14 and the camera assembly 2. A photo of a fruit fly is taken and sent to a host computer for image recognition. When a virgin fly is detected, the electromagnetic three-way valve 3 remains in its current state, and the fruit fly continues to move towards the virgin fly collection bottle. When a non-virgin fly is detected, the host computer controls the electromagnetic three-way valve 3 to switch the airflow direction, the negative pressure of the negative pressure generator 5 disappears, the elastic bladder 13 resets, and the fruit flies located in front of and behind the detection section 14 are squeezed and fixed. If there are no fruit flies, the passage of the guide tube 1 is directly cut off, and the airflow blows the non-virgin flies of the detection section 14 into the separation tube 16 through the blowing tube 15, and finally into the first fruit fly collection bottle, completing the automatic separation and collection of virgin flies. Then the electromagnetic three-way valve 3 resets, the airflow re-enters the negative pressure generator 5, and the fruit flies continue to crawl.

[0039] In some embodiments, a light source assembly may also be disposed below the detection segment 14 to provide uniform illumination and improve the accuracy of image recognition.

[0040] In some embodiments, the camera assembly 2 further includes an infrared sensor, which automatically triggers the camera to capture images when the infrared sensor detects a fruit fly entering the detection section 14, ensuring that a clear image of the fruit fly is captured.

[0041] In some embodiments, a first outer sleeve 17 is fitted over the guide tube 1. A channel for male fruit flies to pass through is provided between the first outer sleeve 17 and the guide tube 1. A plurality of small holes 18 are provided on a section of the guide tube 1 inside the first outer sleeve 17. The small holes 18 have an elliptical structure, with the major axis between 0.8-1.0 mm and the minor axis between 0.6-0.8 mm. Only male fruit flies are allowed to enter the first outer sleeve 17 through the small holes 18. A plastic film is provided outside the small holes 18, forming a one-way channel. After the fruit flies pass through the small holes 18, the plastic film blocks the small holes 18, preventing the male fruit flies from climbing back into the guide tube 1. The first outer sleeve 17 is provided with a connecting tube 6, and a fourth connector 61 is provided at the end of the connecting tube 6, which is connected to a second fruit fly collection bottle to achieve the separation and collection of male fruit flies.

[0042] In some embodiments, the inner wall of the first mantle 17 is coated with pheromones to attract male fruit flies to enter the first mantle 17 through the small hole 18. The pheromone coating enhances the directional movement of male fruit flies and improves separation efficiency. Before the fruit flies enter the detection section 14, a large number of male fruit flies are screened out, thereby accelerating the screening and collection efficiency. This design not only improves the screening efficiency but also reduces interference between fruit flies.

[0043] In some embodiments, a second outer sleeve 163 is also provided on the separation tube 16, and a male fruit fly channel is provided between the second outer sleeve 163 and the separation tube 16. A pheromone coating is also provided inside the second outer sleeve 163. The separation tube 16 is also provided with a sieve hole for connecting the second outer sleeve 163. The sieve hole has an elliptical structure, with a major axis between 0.9-1.2 mm and a minor axis between 0.7-0.9 mm, slightly larger than the small hole 18. The fruit flies blown into the separation tube 16 are male fruit flies and fertilized female fruit flies. After conception, the female fruit fly's abdomen becomes more rounded, and even if the sieve aperture is enlarged, the female fruit fly cannot pass through. Enlarged sieve apertures, on the other hand, allow larger male fruit flies to pass through, further reducing the waste of male fruit flies. The outside of the sieve aperture is also equipped with a one-way plastic film to prevent male fruit flies from returning. The second outer sleeve 163 is also connected to the second fruit fly collection bottle. The fruit flies in the second fruit fly collection bottle are all male, while the fruit flies in the first fruit fly collection bottle are conceived female fruit flies mixed with a small number of male fruit flies.

[0044] In some embodiments, the host computer includes an image acquisition module, an image preprocessing module, a deep learning recognition model, an inference engine, and a decision and control module: The image acquisition module acquires MJPEG format image frames from the camera component 2 in real time through the V4L2 driver framework; the image preprocessing module scales, crops, normalizes, and converts the color space of the acquired images, processing them into 224x224 pixel RGB format, which is used as input to the recognition model; the deep learning recognition model adopts the MobileNetV3-Small lightweight convolutional neural network, which has been pre-trained on a dataset of fruit fly abdomen images labeled with virgin and non-virgin flies through transfer learning; the inference engine adopts the NCNN framework, which calls the neural network processing unit, loads the recognition model weights, performs inference calculations on the preprocessed images, and outputs a two-dimensional vector, representing the probability that the image content is a virgin fly or a non-virgin fly; the decision and control module performs logical judgments based on the inference results and sends control signals to the electromagnetic three-way valve 3 through the GPIO interface.

[0045] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A device for collecting virgin flies for genetic research in fruit flies, characterized in that, include: A guide tube, wherein two elastic bladders are provided on the guide tube and a preset interval is provided between the two elastic bladders as a detection section; The detection section is a transparent structure, and the elastic bladder expands under natural conditions, compressing the inner diameter of the guide tube; A camera assembly is arranged opposite to the detection section and is used to take pictures of fruit flies located in the detection section. A host computer is electrically connected to the camera assembly and is used to perform image recognition on the fruit fly pictures taken by the camera assembly to determine whether they are virgin flies. An air blowing tube and a separation tube are respectively installed on both sides of the detection section, and a one-way valve is installed at the proximal end of the separation tube; An electromagnetic three-way valve has an inlet connected to an air supply module, a first exhaust port connected to the air blowing pipe, and a second exhaust port connected to a negative pressure generator via a pipeline. The negative pressure interface of the negative pressure generator is connected to the two elastic bladders via a negative pressure pipeline. The electromagnetic three-way valve is connected to the host computer and is used to control the airflow through the negative pressure generator to cause the elastic bladders to contract, or to switch the airflow to the air blowing pipe to blow non-virgin flies in the detection section into the separation pipe.

2. The apparatus according to claim 1, characterized in that, The guide tube has a first connector at its proximal end and a second connector at its distal end. The first connector is used to connect to an incubation bottle, and the second connector is used to connect to a virgin fly collection bottle. The separation tube has a third connector at its distal end for connecting to a first fruit fly collection bottle.

3. The apparatus according to claim 1, characterized in that, The guide tube is fitted with a first outer sleeve, forming a male fruit fly passage between the first outer sleeve and the guide tube; a number of small holes are opened on a section of the guide tube inside the first outer sleeve, which only allow male fruit flies to pass through; the first outer sleeve is provided with a connecting tube, the end of which is connected to a second fruit fly collection bottle.

4. The apparatus according to claim 3, characterized in that, A plastic film is provided outside the small hole, forming a one-way channel to prevent fruit flies from returning from the first outer tube to the guide tube.

5. The apparatus according to claim 3, characterized in that, The small hole has an elliptical structure with a major axis of 0.8-1.0 mm and a minor axis of 0.6-0.8 mm.

6. The apparatus according to claim 3, characterized in that, The inner wall of the first mantle is coated with a pheromone coating to attract male fruit flies to enter the first mantle through the small hole.

7. The apparatus according to claim 1, characterized in that, The separating tube is also fitted with a second outer tube, and a male fruit fly passage is provided between the second outer tube and the separating tube; the separating tube has sieve holes for connecting the second outer tube.

8. The apparatus according to claim 1, characterized in that, The inner diameter of the guide tube is 1.5-2.0 mm.

9. The apparatus according to claim 1, characterized in that, The camera assembly also includes an infrared sensor, which automatically triggers the camera assembly to take pictures when the infrared sensor detects a fruit fly entering the detection section.

10. A method for collecting virgin flies using the apparatus according to any one of claims 2-9, characterized in that, Includes the following steps: S1: Install the first connector to the incubation bottle, the second connector to the virgin fly collection bottle, and the third connector to the first fruit fly collection bottle. Place food to attract fruit flies in the virgin fly collection bottle, and arrange the guide tube upward or tilted upward. S2: Open the air supply module, and the airflow enters the negative pressure generator through the electromagnetic three-way valve, causing the elastic bladder to contract, and the fruit fly passes through the detection section; S3: The camera component captures images of fruit flies entering the detection section and sends them to the host computer, which then performs image recognition. S4: When the identification result is a virgin fly, maintain the current state of the electromagnetic three-way valve, and the fruit fly continues to move forward to the virgin fly collection bottle; S5: When the identification result is non-virgin flies, the host computer controls the electromagnetic three-way valve to switch the airflow direction, the negative pressure of the negative pressure generator disappears, the elastic bladder resets to fix the fruit flies on the front and back sides of the detection section, and the airflow blows the non-virgin flies in the detection section into the separation tube through the blowing pipe, and finally into the first fruit fly collection bottle. S6: After the blow-off is completed, the electromagnetic three-way valve is reset, the airflow re-enters the negative pressure generator, and the fruit fly resumes crawling.

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