CN210700168U - Detachable plankton micro-fluidic chip for high-power microscopic image acquisition - Google Patents
Detachable plankton micro-fluidic chip for high-power microscopic image acquisition Download PDFInfo
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- CN210700168U CN210700168U CN201921026161.2U CN201921026161U CN210700168U CN 210700168 U CN210700168 U CN 210700168U CN 201921026161 U CN201921026161 U CN 201921026161U CN 210700168 U CN210700168 U CN 210700168U
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Abstract
The utility model discloses a detachable plankton micro-fluidic chip for high power microscopic image gathers, including the cover plate, substrate and anchor clamps, anchor clamps include clamp and lower clamp, it is equipped with the light trap to go up clamp central authorities, be equipped with down the light trap on the lower clamp, be equipped with more than one detecting channel on the substrate, detecting channel's intermediate part is the detection section, each detecting channel's detection section is located the upper surface of substrate, each detecting channel's entry and export are connected with the joint respectively, lower clamp, the substrate, the supreme range upon range of laying in proper order is down followed to cover plate and last clamp, the thickness of cover plate is 0.03-0.5mm, the cover plate seals to laminate on the substrate, go up clamp and lower clamp fixed connection, the cover plate is fixed in on the substrate through anchor clamps, each detecting channel's detection section is located between light trap and the lower light trap. The chip has the advantages of simple structure, easy assembly and disassembly, easy cleaning and repeated use, and can be used for carrying out high-precision microscopic observation and identification and high-power microscopic image acquisition work on samples.
Description
Technical Field
The utility model belongs to the technical field of plankton, cell detection, concretely relates to a detachable plankton micro-fluidic chip for high-power microscopic image gathers, this chip is applicable to the precision microscope that develops plankton, cell sample and observes and high-power microscopic image gathers.
Background
In recent years, the phenomenon of ecological disasters represented by algal bloom in China gradually emerges, wherein the algal bloom in water areas such as Taihu lake, nested lake and Dian lake becomes a frequent situation. The occurrence of water bloom not only causes serious damage to a water ecosystem, but also seriously affects the health of the surrounding people and the sustainable development of social economy. The rapid monitoring technology of the algal bloom is greatly improved, the international latest technology in the field of water ecology monitoring is tracked, and the intelligent monitoring research on the algal bloom is urgently developed.
At present, the algal bloom monitoring technology at home and abroad mainly comprises a microscopic identification counting method and online instrument monitoring. At present, the microscope identification method widely adopted in China has low efficiency, wastes time and labor (the steps of collecting, fixing, concentrating, pretreating and the like of a water sample for plankton detection are at least 2 days), and is difficult to adapt to the requirement of rapid early warning of the bloom. On the other hand, the existing algae online monitoring instrument is complex and heavy in design, is difficult to adapt to complex and severe field water environment, is mainly suitable for ocean cruise monitoring, is limited in the use of carrying out field monitoring on fresh water bodies with complex conditions such as rivers, lakes and reservoirs and the like, and cannot completely meet the field monitoring requirement.
Recently, due to the development of micro-electro-mechanical processing technology, a lab-on-a-chip (Microfluidics) with a microfluidic chip as a core has become one of the most advanced scientific and technological fields, and compared with the traditional detection technology, the method has the advantages of high efficiency, rapidness, small sample consumption, easiness in realizing automation and portability of instruments, easiness in integrating with the internet of things and the like, and is considered as the development direction of the next-generation monitoring technology. The application technology in the field of water ecology monitoring can be integrated into a micro-fluidic chip platform, and the monitoring of the micro-fluidic chip for plankton has great value.
However, since the individual size of algae, zooplankton, etc. in the water is small, mostly in the order of micron size, it is observed only by using high power microscope. In the current manufacturing technology of the microfluidic chip, the thickness of the cover plate can only be 1-2 mm. The excessive thickness of the cover plate of the microfluidic chip brings a series of problems. Among them, the most important disadvantages are that it is difficult to use a high power objective lens with short working distance for observation, and to perform high magnification precision optical observation and high power microscopic image acquisition. Therefore, in the development of a microfluidic chip for plankton detection, it is necessary to reduce the thickness of a chip cover plate as much as possible.
In addition, due to the complexity of the field water body, the collected water sample often contains a large amount of non-biological impurities, the chip bonded at one time is used for detection, the processing cost of the chip is high, the blockage of a chip detection channel is easy to occur, and the chip is scrapped due to the huge cleaning difficulty.
Disclosure of Invention
In order to solve the problems existing in the prior art, the utility model provides a detachable plankton micro-fluidic chip for high-power microscopic image acquisition, this chip simple structure easily assembles and dismantles, easily washs, can use repeatedly, and can be used to develop the microscope observation appraisal of high accuracy, high-power microscopic image acquisition work to the sample.
Realize the utility model discloses the technical scheme that above-mentioned purpose adopted does:
the utility model provides a detachable plankton micro-fluidic chip for high power shows collection of micro-image, including the cover plate, the substrate with draw together anchor clamps, anchor clamps include clamp and lower clamp, it is equipped with the light trap to go up clamp central authorities, be equipped with down the light trap on the clamp down, be equipped with more than one detection channel on the substrate, if detection channel is many, many detection channel arrange side by side, detection channel's mid portion is the detection section, the detection section of each detection channel is located the upper surface of substrate, lower clamp, the substrate, cover plate and last clamp from supreme range upon range of laying in proper order down, the thickness of cover plate is 0.03-0.5mm, the cover plate seals up on the substrate, the detection section of each detection channel of cover plate seal, go up clamp and lower clamp fixed connection, the cover plate is fixed in on the substrate through anchor clamps, the detection section of each detection channel is located between light trap and the lower light trap.
The upper clamp body and the lower clamp body are both plate-shaped, a plurality of through holes are uniformly distributed around the upper light transmission hole, the through holes are uniformly distributed at corresponding positions around the lower light transmission hole, and the upper clamp body and the lower clamp body are connected through bolts.
The upper clamp body comprises an upper metal shell and an upper padding plate, the upper metal shell and the upper padding plate are identical in shape, the upper padding plate is bonded in the upper metal shell, the lower shell comprises a lower metal shell and a lower padding plate, the lower metal shell and the lower padding plate are identical in shape, the lower padding plate is bonded in the lower metal shell, the upper padding plate is attached to the cover plate, and the lower padding plate is attached to the substrate.
The upper light hole and the lower light hole are square or circular.
The detection channel is of a symmetrical structure and is in a shape like a Chinese character 'ji', the detection channel is composed of two connecting sections and a detection section, the two side parts of the detection channel are respectively the connecting sections, one end of each connecting section is respectively connected with the two ends of the detection section, the two connecting sections of each detection channel are respectively positioned in the substrate, and the other end of each connecting section of each detection channel is respectively connected with a joint.
The substrate and the cover plate are both square, the projection of each detection channel on a plane parallel to the substrate is in a shape of a Chinese character 'yi', the detection section of each detection channel is parallel to the edge of the upper surface of the substrate in the length direction, and the inlet and the outlet of each detection channel are respectively positioned on two side walls in the width direction of the substrate.
The detection channel is of a symmetrical structure and is U-shaped, the detection channel is composed of two connecting sections and a detection section, the two side parts of the detection channel are respectively the connecting sections, one ends of the two connecting sections are respectively connected with the two ends of the detection section, the two connecting sections of each detection channel are respectively positioned in the substrate, and the other ends of the two connecting sections of each detection channel are respectively connected with a joint.
The substrate and the cover plate are both square, the projection of each detection channel on a plane parallel to the substrate is in a shape of a Chinese character 'yi', the detection section of each detection channel is parallel to the edge of the length direction of the upper surface of the substrate, and the inlet and the outlet of each detection channel are respectively positioned on the lower surface of the substrate.
The detection channel is of a symmetrical structure and comprises two connecting sections and detection sections, the two side parts of the detection channel are respectively the connecting sections, one ends of the two connecting sections are respectively connected with the two ends of the detection section, the connecting sections are U-shaped, the two connecting sections of each detection channel are respectively positioned in the substrate, and the other ends of the two connecting sections of each detection channel are respectively connected with a joint.
The substrate and the cover plate are both square, the projection of each detection channel on a plane parallel to the substrate is in a shape of a Chinese character 'yi', the detection section of each detection channel is parallel to the edge of the length direction of the upper surface of the substrate, and the inlet and the outlet of each detection channel are respectively positioned on the upper surface of the substrate.
Compared with the prior art, the beneficial effects and advantages of the utility model reside in that:
1. the utility model discloses an anchor clamps adopt special design, under the difficult broken circumstances of assurance cover plate, firmly fix on the cover plate, the existence of anchor clamps simultaneously can reduce the thickness of cover plate by a wide margin for can use the low price, thickness is only 0.13 mm's common microscope cover glass as the cover plate, thereby can adopt the high power objective of various short working distance such as 40 times on the biological microscope, 100 times to observe, be 1-2 mm's conventional micro-fluidic chip for cover plate thickness, the utility model discloses the at utmost has avoided the cover plate to cross thick to the sample to observe a series of negative effects such as chromatic aberration change, luminousness decline, image deformation that bring.
2. The utility model discloses a detection object of chip is for being rich in the open-air water sample of various impurity, inorganic particulate matter, plankton, and the test passage of chip changes the jam for ordinary sample. If adopt the chip of the unable separation of cover plate, the substrate of conventional adoption bonding technology preparation, the washing of chip is unusual difficult, very easily scraps, to this problem, the utility model discloses a chip design is detachable, the structure of using repeatedly, and the part simple structure of adoption, dismantlement simple to operate very easily wash, can use repeatedly, chip abluent difficult problem when effectively having solved the open-air water sample of monitoring.
3. The utility model discloses a chip simple structure, the spare part that adopts is common consumptive material, and the cost of preparation, use and maintenance is low, is suitable for large-scale production and popularization and application.
Drawings
Fig. 1 is an exploded view of a detachable plankton microfluidic chip for high-power microscopic image acquisition.
Fig. 2 is a schematic structural diagram of a detachable plankton microfluidic chip for high-power microscopic image acquisition.
Fig. 3 is a schematic structural view of the upper clamp body.
Fig. 4 is a cross-sectional view of a substrate of example 1.
Fig. 5 is a cross-sectional view of a substrate of example 1.
Fig. 6 is a cross-sectional view of a substrate of example 1.
Fig. 7 is a cross-sectional view of a substrate of example 2.
Fig. 8 is a cross-sectional view of a substrate of example 3.
Fig. 9 is a cross-sectional view of a substrate of example 3.
The structures of the detection channels in fig. 4, 5 and 6 are different, and the structures of the detection channels in fig. 8 and 9 are different.
Wherein, 1-cover plate, 2-substrate, 3-upper clip, 4-lower clip, 5-upper light hole, 6-detection channel, 7-detection section, 8-connecting section, 9-joint, 10-perforation, 11-bolt, 12-upper metal shell, 13-upper backing plate, 14-lower light hole.
Detailed Description
The present invention will be described in detail with reference to the accompanying drawings.
Example 1
The structure of the detachable plankton microfluidic chip for high-power microscopic image acquisition provided in this example 1 is shown in fig. 1 and 2, and includes a cover plate, a substrate and a clamp.
The thickness of the cover plate 1 is 0.03-0.05mm, the cover plate can be made of glass, toughened glass, quartz and other materials, for example, a cover glass with the thickness of 0.13mm used for conventional microscope observation, the cover plate 1 is square, and the side length of the cover plate is 3.0 cm.
The substrate 2 can be made of glass, toughened glass, quartz, organic glass, plastic, polydimethylsiloxane, polymethyl methacrylate, polycarbonate and the like. The substrate 2 is rectangular, and the length of the substrate 2 is 7.5cm, the width is 3.0cm, and the thickness is 2.0 mm.
The substrate 2 is provided with 2 detection channels 6 which are arranged side by side, the cross section of each detection channel 6 is square, the depth is 20 mu m-5 mm, the width is 50 mu m-10 mm, and the width of each detection channel is larger than the depth of each detection channel. As shown in fig. 4, 5 and 6, the detection channel 6 is a symmetrical structure, the detection channel 6 is in a shape of a Chinese character 'ji', the detection channel 6 is composed of two connection sections 8 and a detection section 7, and the two connection sections 8 are respectively connected with two ends of the detection section 7. The detection section 7 of each detection channel 6 is located on the upper surface of the substrate 2, and the two connection sections 8 of each detection channel 6 are located in the substrate respectively. The projection of each detection channel 6 on the plane parallel to the substrate 2 is in a shape of a Chinese character 'yi', the detection section 7 of each detection channel 6 is parallel to the edge of the upper surface of the substrate 2 in the length direction, the inlet and the outlet of each detection channel 6 are respectively positioned on two side walls of the substrate 2 in the width direction, and the inlet and the outlet of each detection channel 6 are respectively connected with a connector 9.
When detecting different types of plankton, the depth of the detection channel is selected and adjusted according to the size of the plankton, and the depth of the selected detection channel is larger than the size of the plankton to be detected.
The clamp comprises an upper clamp body 3 and a lower clamp body 4, the upper clamp body 3 and the lower clamp body 4 are the same in shape and size, and the upper clamp body 3 and the lower clamp body 4 are both plate-shaped.
As shown in fig. 3, the upper clamp 3 is composed of an upper metal case 12 and an upper pad 13, the upper metal case 12 and the upper pad 13 have the same shape, and the upper pad 13 is bonded to the inside of the upper metal case 12. The upper metal shell 12 is made of aluminum, copper, stainless steel, titanium, alloy and the like, and the upper backing plate 13 is made of rubber, silica gel, plastic, polytetrafluoroethylene and the like. The center of the upper clamp body 3 is provided with a square upper light hole 5, and the side length of the upper light hole 5 is 2.0 cm. The outer sides of four corners of the upper light-transmitting hole 5 are respectively provided with a through hole 10, the four through holes 10 are distributed in a square shape, the center of the square and the center of the upper light-transmitting hole 10 are on the same straight line, and the distance between every two adjacent through holes is 3.5 cm.
The lower clamp body is composed of a lower metal shell and a lower backing plate, the shapes of the lower metal shell and the lower backing plate are the same, and the lower backing plate is bonded in the lower metal shell. The lower metal shell is made of aluminum, copper, stainless steel, titanium, alloy and the like, and the lower backing plate is made of rubber, silica gel, plastic, polytetrafluoroethylene and the like. The center of the lower clamp body is provided with a square lower light hole 14, and the side length of the lower light hole 14 is 2.0 cm. The outer sides of four corners of the lower light-transmitting hole 14 are respectively provided with through holes 10, the four through holes 10 are distributed in a square shape, the center of the square and the center of the upper light-transmitting hole are on the same straight line, and the distance between every two adjacent through holes is 3.5 cm.
The use method of the detachable plankton microfluidic chip for collecting the high-power microscopic image comprises the following steps:
preparation before assembly
1. Cleaning: placing all the parts in an ultrasonic cleaning instrument (Shanghai Lele ZL6-180A), adding a detergent, ultrasonically cleaning the parts such as the substrate, the cover plate, the clamp, the joint and the like for 3-15min, then cleaning for 1-2 times with ultrapure water for 3-15min each time, blow-drying, if the detection channel is blocked, extracting the detergent by using an injector, and repeatedly washing the detection channel 6.
2. Coating a sealant: uniformly coating waterproof sealing materials such as vaseline, lubricating oil, lubricating grease, vacuum silicone grease, hot melting wax, hot melting glue and the like on the upper surface of the substrate, and taking care to avoid coating the waterproof sealing materials into a detection channel;
b, chip assembly:
1. placing a lower clamp on a clean workbench, enabling a lower gasket to face upwards, enabling one surface of a substrate, which is provided with detection channels, to face upwards and placing the surface of the substrate on the lower gasket, carefully placing a cover plate on the substrate, paying attention to the placement position of a cover plate, enabling detection sections of the detection channels to be covered by the cover plate, slightly pressing the cover plate, paying attention to avoiding bubbles between the cover plate and the substrate, placing an upper clamp on the cover plate, enabling the upper gasket to be attached to the cover plate, and adjusting the position of the upper clamp to correspond to the position of the lower clamp;
2. the four bolts are respectively arranged at the four corners of the clamp, each bolt is gradually screwed, and the cover plate is prevented from being broken due to over-tightening of the bolts;
3. adhering the joint on the substrate by using hot melt adhesive, hot melt wax and the like to connect the joint with the inlet and the outlet of the detection channel;
c chip sealing performance test
Connecting the inlet of each detection channel with an injector filled with water solution through a pipeline, pushing the injector, pushing the water solution to pass through each detection channel under the atmospheric pressure of 0.5-3, continuing the whole test process for 0.5-5min, detecting whether the chip leaks water, testing the sealing performance of the chip, if the chip does not leak water, performing sample detection, and if not, detaching and reinstalling.
D, sample detection:
placing the installed microfluidic chip on an objective table of an optical microscope (an Olympus CX31 biological microscope, and a matched CCD model is Yinmei Jing DFK 33UX249) provided with a CCD, clamping the microfluidic chip by using an objective table pressing sheet, aligning a detection channel to be observed with a light through hole, turning on a microscope light source, adjusting a microscope light path, selecting an objective lens, adjusting a focal plane by adjusting a coarse focusing screw and a fine focusing screw to enable the microscope to obtain the clearest image, and adjusting a microscope stage to enable the detection channel to be observed by the microfluidic chip to be in the center of the field of view of the microscope;
connecting the inlet of the detection channel to be observed with the water sample of plankton to be detected through a pipeline, under the power drive of an injection pump (venturi WH-SP-02 type), allowing the water sample containing plankton to be detected to pass through the detection channel of the chip at a constant speed at a known fixed flow rate, selecting the detection channel with a small depth (the depth of the detection channel is 50 μm) to carry out micro plankton observation and detection, wherein the flow rate range of water sample injection is 0.05-2.0ml/min, and when selecting the detection channel with a large depth (the depth of the detection channel is 250 μm) to carry out micro plankton observation and detection, the flow rate range of water sample injection is 0.5-30 ml/min;
the method comprises the steps of observing and recording the types and the quantity of various plankton passing through a chip detection channel in unit time in the field of view of a microscope at present, calculating the actual volume of a water sample observed and recorded according to the sectional area of the detection channel, the sample introduction speed of the water sample and the unit time of detection, and calculating the actual types and the quantity of various plankton in the water sample to be detected according to data.
Meanwhile, the sample to be detected can acquire the image into a computer for storage through a CCD (charge coupled device) connected with an optical microscope at a fixed image acquisition frame rate (1-60 times/second) and within a resolution range (800 × 600-.
Example 2
The difference between this embodiment and embodiment 1 lies in the shape of the detection channel, in this embodiment, as shown in fig. 7, the detection channel is U-shaped, the detection section of each detection channel is located on the upper surface of the substrate, and the two connection sections of each detection channel are located in the substrate respectively. The projection of each detection channel in the direction parallel to the substrate is in a straight shape, the detection section of each detection channel is parallel to the edge of the length direction of the upper surface of the substrate, and the inlet and the outlet of each detection channel are respectively positioned on the lower surface of the substrate.
Example 3
The present embodiment is different from embodiment 1 in the shape of the detection channel, and in this embodiment, as shown in fig. 8 and 9, the connection section is U-shaped, the detection section of each detection channel is located on the upper surface of the substrate, and the two connection sections of each detection channel are located in the substrate, respectively. The projection of each detection channel in the direction parallel to the substrate is in a shape of a Chinese character 'yi', the detection section of each detection channel is parallel to the edge of the length direction of the upper surface of the substrate, and the inlet and the outlet of each detection channel are respectively positioned on the upper surface of the substrate.
Claims (10)
1. The utility model provides a detachable plankton micro-fluidic chip for high power microscopic image gathers, includes cover plate and substrate, its characterized in that: the substrate detection device is characterized by further comprising a clamp, the clamp comprises an upper clamp body and a lower clamp body, the central portion of the upper clamp body is provided with an upper light hole, the lower clamp body is provided with more than one detection channel, when the detection channels are multiple, the multiple detection channels are arranged side by side, the middle portion of each detection channel is a detection section, the detection sections of the detection channels are located on the upper surface of the substrate, the lower clamp body, the substrate, the cover plate and the upper clamp body are sequentially stacked from bottom to top, the thickness of the cover plate is 0.03-0.5mm, the cover plate is sealed and attached to the substrate, the detection sections of the detection channels are sealed by the cover plate, the upper clamp body and the lower clamp body are fixedly connected, the cover plate is fixed on the substrate through the clamp, and the detection sections of the detection channels are located between the upper light hole and the lower.
2. The detachable plankton microfluidic chip for high-power microscopic image acquisition according to claim 1, characterized in that: the upper clamp body and the lower clamp body are both plate-shaped, a plurality of through holes are uniformly distributed around the upper light transmission hole, the through holes are uniformly distributed at corresponding positions around the lower light transmission hole, and the upper clamp body and the lower clamp body are connected through bolts.
3. The detachable plankton microfluidic chip for high-power microscopic image acquisition according to claim 2, characterized in that: the upper clamp body comprises an upper metal shell and an upper padding plate, the upper metal shell and the upper padding plate are identical in shape, the upper padding plate is bonded in the upper metal shell, the lower shell comprises a lower metal shell and a lower padding plate, the lower metal shell and the lower padding plate are identical in shape, the lower padding plate is bonded in the lower metal shell, the upper padding plate is attached to the cover plate, and the lower padding plate is attached to the substrate.
4. The detachable plankton microfluidic chip for high-power microscopic image acquisition according to claim 2, characterized in that: the upper light hole and the lower light hole are square or circular.
5. The detachable plankton microfluidic chip for high-power microscopic image acquisition according to claim 2, characterized in that: the detection channel is of a symmetrical structure and is in a shape like a Chinese character 'ji', the detection channel is composed of two connecting sections and a detection section, the two side parts of the detection channel are respectively the connecting sections, one end of each connecting section is respectively connected with the two ends of the detection section, the two connecting sections of each detection channel are respectively positioned in the substrate, and the other end of each connecting section of each detection channel is respectively connected with a joint.
6. The detachable plankton microfluidic chip for high-power microscopic image acquisition according to claim 5, wherein: the substrate and the cover plate are both square, the projection of each detection channel on a plane parallel to the substrate is in a shape of a Chinese character 'yi', the detection section of each detection channel is parallel to the edge of the upper surface of the substrate in the length direction, and the inlet and the outlet of each detection channel are respectively positioned on two side walls in the width direction of the substrate.
7. The detachable plankton microfluidic chip for high-power microscopic image acquisition according to claim 2, characterized in that: the detection channel is of a symmetrical structure and is U-shaped, the detection channel is composed of two connecting sections and a detection section, the two side parts of the detection channel are respectively the connecting sections, one ends of the two connecting sections are respectively connected with the two ends of the detection section, the two connecting sections of each detection channel are respectively positioned in the substrate, and the other ends of the two connecting sections of each detection channel are respectively connected with a joint.
8. The detachable plankton microfluidic chip for high-power microscopic image acquisition according to claim 7, wherein: the substrate and the cover plate are both square, the projection of each detection channel on a plane parallel to the substrate is in a shape of a Chinese character 'yi', the detection section of each detection channel is parallel to the edge of the length direction of the upper surface of the substrate, and the inlet and the outlet of each detection channel are respectively positioned on the lower surface of the substrate.
9. The detachable plankton microfluidic chip for high-power microscopic image acquisition according to claim 2, characterized in that: the detection channel is of a symmetrical structure and comprises two connecting sections and detection sections, the two side parts of the detection channel are respectively the connecting sections, one ends of the two connecting sections are respectively connected with the two ends of the detection section, the connecting sections are U-shaped, the two connecting sections of each detection channel are respectively positioned in the substrate, and the other ends of the two connecting sections of each detection channel are respectively connected with a joint.
10. The removable plankton microfluidic chip for high-power microscopic image acquisition according to claim 9, wherein: the substrate and the cover plate are both square, the projection of each detection channel on a plane parallel to the substrate is in a shape of a Chinese character 'yi', the detection section of each detection channel is parallel to the edge of the length direction of the upper surface of the substrate, and the inlet and the outlet of each detection channel are respectively positioned on the upper surface of the substrate.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110252436A (en) * | 2019-07-03 | 2019-09-20 | 水利部中国科学院水工程生态研究所 | A kind of detachable planktonic organism micro-fluidic chip for high power micro image collection |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110252436A (en) * | 2019-07-03 | 2019-09-20 | 水利部中国科学院水工程生态研究所 | A kind of detachable planktonic organism micro-fluidic chip for high power micro image collection |
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