Method for monitoring movement, sleep and circadian rhythm of fruit flies

Abstract

The invention relates to the field of insect behavioristics, in particular to a method for continuously monitoring movement, sleep and circadian rhythm of diptera insect fruit flies in real time, which comprises the following steps: S1, preparing fruit fly food; s2, taking an acrylic tube A with holes in two ends, adding 2cm of solid food into one end of the acrylic tube A, putting the fruit flies into the acrylic tube A, and plugging the other end of the acrylic tube A with a 2cm tampon; s3, two acrylic pipes B with holes formed in the two ends are arranged in the acrylic double-opening pipe A in a sleeving mode, and an A and B combined device is formed; s4, screwing the A and B combined device into an acrylic double-head pipe C to form an A, B and C combined device; s5, putting the A, B and C combined device into an LAM25 monitor for monitoring; and S6, analyzing the data through software for sleep and circadian rhythm analysis. On the basis of an original insect behavior monitoring system, a research object is excluded to the species of the fruit flies, a monitoring device is optimized according to monitoring data, and an accurate method for monitoring movement, sleep and circadian rhythm states of the fruit flies is established.

Description

Technical Field

[0001] The present invention relates to the field of insect behavior, and in particular to a method and research system for continuously monitoring fruit fly movement, sleep and circadian rhythm Background Art

[0002] Using LAM25 (Locomotor Activity Monitor 25) to monitor insect behavior is one of the current monitoring methods for larger insects. The device was invented and manufactured by TriKinetics, an American company, and has been widely used in insect behavior monitoring.

[0003] Each LAM25 monitor can measure the independent movement behavior of 32 insect samples at a time, with each insect placed in its own tube. When the insect moves freely back and forth in the tube, it will block the infrared beam passing through the middle of the tube, and this data will be automatically recorded by the computer as the insect's behavioral parameters. In a typical experiment, food is placed at the bottom of the tube, followed by a single insect, and then the tube mouth is plugged with cotton; 32 tubes are inserted into the corresponding holes of the monitor for behavioral monitoring. During the entire experiment, the behavior of all 32 insect individuals can be continuously monitored for days or weeks, and data will be uploaded to the computer at intervals (usually every minute). These detailed records can accurately measure the patterns of insect activity: the distribution and intensity of movement behavior, sleep, and related circadian rhythm indicators.

[0004] The LAM25 monitor is made of a transparent polycarbonate plastic that can be penetrated by light at any point in the tube. It is compact, lightweight, and allows air to circulate in the plastic tube when placed in the behavior monitoring tube. The perforated design can accommodate tubes of any length. An on / off visible light sensor is installed in the monitor to record the light status of the surrounding environment in real time during behavior monitoring. This light status data is stored together with the corresponding behavior parameters, which is very helpful for analyzing the insect's circadian rhythm and adaptability to external stimulus light.

[0005] The original device without modification has many limitations when used for insect monitoring, which leads to the deviation of the monitoring results from the actual insect activities, such as: 1. The monitored insects are small. If the insects only move on one side, the number of times the infrared rays are blocked will be reduced, affecting the accuracy of the monitoring results; 2. The movement ability of different insects varies greatly. The same movement space is suitable for some insects with large movement, but it is impossible to obtain accurate movement and sleep information for insects with less movement. Based on the existing LAM25 insect behavior monitoring, the present invention improves the size of the insect monitoring tube and other conditions, and establishes a method for monitoring fruit fly behavior, sleep and circadian rhythm. Summary of the invention

[0006] The purpose of the present invention is to solve the limitations of the prior art and propose a method for monitoring the movement, sleep and circadian rhythm of fruit flies to solve the problem that the LAM25 monitor cannot accurately monitor the circadian rhythm of fruit flies.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] A method for monitoring fruit fly movement, sleep and circadian rhythm, comprising the following specific steps:

[0009] S1: Fruit fly food preparation: To ensure the health of fruit flies during monitoring, the following three types of food were used to meet the experimental conditions, including solid food a, solid food b, and liquid food c;

[0010] S2: Take a double-ended open-hole acrylic tube A, add 2 cm of solid food at one end, put in fruit flies, and plug the other end with a 2 cm cotton plug;

[0011] S3: Take two acrylic double-opening tubes B, insert the two double-opening acrylic tubes B into the acrylic double-opening tube A, keep one acrylic double-opening tube B at each end of the double-opening acrylic tube A, and form an A, B combination device;

[0012] S4: Take the acrylic double-opening tube C, screw the A and B combination device into the acrylic double-opening tube C, and form the A, B, C combination device;

[0013] S5: Place the A, B, and C combination into the LAM25 monitor to monitor sleep under LD = 12:12 or 14:10 conditions and monitor the circadian rhythm of fruit flies under DD conditions;

[0014] S6: Analyze the data using sleep and circadian rhythm analysis software.

[0015] As a further technical solution of the present invention, in S1, the solid food a is a sterilized solid food of honey and agar, the concentration of honey is 10%, and the concentration of agar is 1%.

[0016] As a further technical solution of the present invention, in S1, the solid food b is a sterilized solid food of sucrose and agar, the concentration of sucrose is 10%, and the concentration of agar is 1%.

[0017] As a further technical solution of the present invention, in S1, the liquid food c is a liquid culture medium of honey water, and the concentration of the honey water is 10%.

[0018] As a further technical solution of the present invention, in S2, the dimensions of the double-ended perforated acrylic tube A are 1 cm in inner diameter, 1.4 cm in outer diameter, and 10 cm in length, and the movement distance of the fruit flies in the double-ended perforated acrylic tube A is 6 cm.

[0019] As a further technical solution of the present invention, in S3, the size of the acrylic double-opening tube B is an inner diameter of 1.4 cm and an outer diameter of 2.2 cm.

[0020] As a further technical solution of the present invention, in S4, the size of the acrylic double-opening tube C is 2.2 cm in inner diameter and 2.5 cm in outer diameter.

[0021] As a further technical solution of the present invention, in S4, the double-opened acrylic tube A is placed in the center of the acrylic double-opening tube C through the acrylic double-opening tube B using a sleeve mode.

[0022] The advantages of the present invention are that, based on the original insect behavior monitoring system, the research object is expanded to the fruit fly species, and the monitoring device is optimized according to the monitoring data, thereby establishing a more accurate monitoring method for monitoring the sleep and circadian rhythm of the fruit fly. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 Schematic diagram of the process of assembling three types of acrylic tubes A, B, and C into a casing system and monitoring them;

[0024] Figure 2 A comparison of the sleep and circadian rhythm data of Drosophila monitored using the cannula device and the dedicated monitor DAM2;

[0025] Figure 3 A comparison chart of using the tube device to monitor fruit flies and using the LAM25 device to monitor fruit fly movement, sleep and circadian rhythm;

[0026] Figure 4 The present invention provides a flow chart of a method for monitoring fruit fly movement, sleep and circadian rhythm. DETAILED DESCRIPTION

[0027] In order to make the technical means, creative features, objectives and effects achieved by the present invention easy to understand, the present invention is further explained below in conjunction with specific implementation methods.

[0028] Please see attached Figure 4 , a method for monitoring fruit fly movement, sleep and circadian rhythm, comprising the following specific steps:

[0029] S1: Fruit fly food preparation: To ensure the health of fruit flies during monitoring, the following three types of food were used to meet the experimental conditions, including solid food a, solid food b, and liquid food c;

[0030] Solid food a is a sterilized solid food of honey and agar, with the concentration of honey being 10% and the concentration of agar being 1%;

[0031] Solid food b is a sterilized sucrose and agar solid food, the concentration of sucrose is 10%, and the concentration of agar is 1%;

[0032] Liquid food c is a liquid culture medium of honey water, and the concentration of honey water is 10%.

[0033] S2: Take a double-ended open-hole acrylic tube A, add 2 cm of solid food at one end, put in fruit flies, and plug the other end with a 2 cm cotton plug;

[0034] The dimensions of the double-ended perforated acrylic tube A are an inner diameter of 1 cm, an outer diameter of 1.4 cm, and a length of 10 cm. The movement distance of the fruit flies in the double-ended perforated acrylic tube A is 6 cm.

[0035] S3: Take two acrylic double-opening tubes B, insert the two double-opening acrylic tubes B into the acrylic double-opening tube A, keep one acrylic double-opening tube B at each end of the double-opening acrylic tube A, and form an A, B combination device;

[0036] The dimensions of the acrylic double-opening tube B are 1.4 cm in inner diameter and 2.2 cm in outer diameter.

[0037] S4: Take the acrylic double-opening tube C, and screw the A and B assembly into the acrylic double-opening tube C, that is, use the sleeve mode to put the double-opening acrylic tube A through the acrylic double-opening tube B and place it in the center of the acrylic double-opening tube C to form the A, B, C assembly;

[0038] The dimensions of the acrylic double-opening tube C are 2.2cm inner diameter and 2.5cm outer diameter.

[0039] S5: Place the combination of A, B, and C into a LAM25 monitor to monitor sleep under LD=12:12 or 14:10 conditions, and monitor the endogenous circadian rhythm of fruit flies under DD conditions.

[0040] S6: Analyze the data using sleep and circadian rhythm analysis software.

[0041] Example 1

[0042] Before monitoring, in order to verify the reliability of the device, a similar device was used to monitor the movement, sleep and circadian rhythm of fruit flies, and the same results as the fruit fly behavior monitor were obtained. In addition, in order to verify that the method has been improved, the unsheathed device and the sheathed device were used to monitor fruit flies, and it was found that the accuracy of monitoring was effectively improved.

[0043] like Figure 1 As shown: The method for monitoring fruit fly behavior, sleep and circadian rhythm consists of three parts: A, B, and C. When monitoring the target insect, the following steps need to be performed:

[0044] 1. Insect rearing and circadian rhythm acclimation;

[0045] 2. Preparation of food tube A, filling and sealing of insects;

[0046] 3. Combination of A, B and C casing devices;

[0047] 4. Monitoring of behavior;

[0048] 5. Analysis of data.

[0049] like Figure 2 Shown: The reliability of the method was confirmed by comparing the behavior of fruit flies on the LAM25 monitor and DAM2.

[0050] like Figure 3 As shown: The monitoring of fruit flies on the LAM25 monitor using the sleeve device and not using the sleeve device can significantly improve the monitoring accuracy of fruit flies under DD conditions. The data is more consistent with their actual movement conditions, the peak graph is more obvious, and the error is smaller.

[0051] From the above description, it can be seen that the above-mentioned embodiments of the present invention realize the following technical innovations: on the basis of the original insect behavior monitoring system, the research object is specialized to the fruit fly species, and the monitoring device is optimized according to the monitoring data, and a more accurate monitoring method is established for monitoring the sleep and circadian rhythm status of the fruit fly.

[0052] Those skilled in the art should understand that the discussion of any of the above embodiments is merely illustrative and is not intended to imply that the scope of the present invention (including the claims) is limited to these examples. Under the concept of the present invention, the technical features in the above embodiments or different embodiments may be combined, the steps may be implemented in any order, and there are many other variations of the different aspects of the present invention as described above, which are not provided in detail for the sake of simplicity.

[0053] The present invention is intended to cover all such substitutions, modifications and variations that fall within the broad scope of the claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for monitoring fruit fly movement, sleep and circadian rhythm, characterized in that: The specific steps include: S1: Fruit fly food preparation: To ensure the health of fruit flies during monitoring, the following three types of food were used to meet the experimental conditions, including solid food a, solid food b, and liquid food c; S2: Take a double-ended open-hole acrylic tube A, add 2 cm of solid food at one end, put in fruit flies, and plug the other end with a 2 cm cotton plug; S3: Take two acrylic double-opening tubes B, insert the two double-opening acrylic tubes B into the acrylic double-opening tube A, keep one acrylic double-opening tube B at each end of the double-opening acrylic tube A, and form an A, B combination device; S4: Take the acrylic double-opening tube C, screw the A and B combination device into the acrylic double-opening tube C, and form the A, B, C combination device; S5: Place the A, B, and C combination devices into the LAM25 monitor to monitor circadian rhythm and sleep under LD = 12:12 or 14:10 conditions, and monitor the endogenous circadian rhythm and sleep of fruit flies under DD conditions; S6: Analyze the data using sleep and circadian rhythm analysis software.

2. A method for monitoring fruit fly movement, sleep and circadian rhythm according to claim 1, characterized in that: In S1, the solid food a is a sterilized solid food of honey and agar, the concentration of honey is 10%, and the concentration of agar is 1%.

3. A method for monitoring fruit fly movement, sleep and circadian rhythm according to claim 2, characterized in that: In S1, the solid food b is a sterilized solid food of sucrose and agar, wherein the concentration of sucrose is 10% and the concentration of agar is 1%.

4. A method for monitoring fruit fly movement, sleep and circadian rhythm according to claim 3, characterized in that: In the S1, the liquid food c is a liquid culture medium of honey water, and the concentration of the honey water is 10%.

5. A method for monitoring fruit fly movement, sleep and circadian rhythm according to claim 1, characterized in that: In S2, the dimensions of the double-ended perforated acrylic tube A are 1 cm in inner diameter, 1.4 cm in outer diameter, and 10 cm in length, and the movement distance of the fruit flies in the double-ended perforated acrylic tube A is 6 cm.

6. A method for monitoring fruit fly movement, sleep and circadian rhythm according to claim 1, characterized in that: In the above-mentioned S3, the dimensions of the acrylic double-opening tube B are an inner diameter of 1.4 cm and an outer diameter of 2.2 cm.

7. A method for monitoring fruit fly movement, sleep and circadian rhythm according to claim 1, characterized in that: In the above-mentioned S4, the dimensions of the acrylic double-opening tube C are an inner diameter of 2.2 cm and an outer diameter of 2.5 cm.

8. A method for monitoring fruit fly movement, sleep and circadian rhythm according to claim 1, characterized in that: In the S4, the double-end opening acrylic tube A is placed in the center of the acrylic double-opening tube C through the acrylic double-opening tube B using a sleeve mode.

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