A vitrification micro-carrier device for temperature measurement

Abstract

The utility model discloses a kind of vitrification cooling micro sample carrier devices of extension temperature measurement, including the protective sleeve pipe, sample ring, support rod connected in sequence.The protective sleeve pipe is stretchable corrugated structure, fixed on support rod upper end, extend to the sample on the sample ring of outermost edge can be protected;Support rod is located inside protective sleeve pipe, and extend to outside along axial direction, and its end connects sample ring;Sample ring is micro-nano annular structure for carrying trace biological sample.The device structure is simple, heat conduction path is clear, suitable for the positioning fixation and stable support of droplet in vitrification cooling, with good thermal response characteristic and mechanical strength, especially suitable for trace biological material sample loading with temperature measurement demand.

Description

Technical Field

[0001] This invention belongs to the field of low-temperature biological sample preservation technology, and particularly relates to a vitrification cooling micro-sample loading device with expandable temperature measurement function, which is suitable for loading micro biological samples, chemical samples and other samples under extreme low temperature conditions. Background Technology

[0002] In the study of cryopreservation and thawing of biological samples, vitrification cooling has significant advantages in preserving trace samples such as sperm, oocytes, and embryos due to its ability to prevent ice crystal formation and reduce cell damage. However, existing vitrification cooling sample carriers still have many shortcomings: on the one hand, traditional carriers usually only have a single cooling function and are difficult to integrate with temperature monitoring modules, making it impossible to record temperature changes of the sample in real time during freezing and thawing; on the other hand, the large contact area between the carrier and the sample affects the heat exchange efficiency between the sample and the cooling medium, and the protective structure is mostly a closed design, which is difficult to disassemble and assemble in a liquid nitrogen environment, making operation inconvenient. In addition, although open cooling devices are easy to operate, they lack effective mechanical protection for the sample and are susceptible to external thermal disturbances and physical damage.

[0003] Therefore, there is an urgent need to develop a vitrified cooling sample carrier device that is compact, highly protective, has a fast thermal response, and supports temperature monitoring, in order to meet the dual requirements of temperature control accuracy and ease of operation for micro biological samples during extreme low-temperature processing. Utility Model Content

[0004] Purpose of the utility model: In order to overcome the problems of existing vitrification freezing devices and loading technologies for micro-biological samples lacking scalable temperature monitoring functions and only allowing one side of the sample to contact liquid nitrogen, this utility model proposes a vitrification cooling micro-sample loading device with extended temperature measurement. It has a compact structure, strong protection, and good thermal response and mechanical properties, and is especially suitable for freezing and rewarming of micro-biological materials with temperature measurement requirements.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] An extended temperature measurement vitrified cooling microsample carrier device, characterized in that it includes:

[0007] The support rod is in the shape of an inverted step, used to prevent liquid nitrogen from escaping;

[0008] A sample carrying ring, located at the end of the support rod, is used to carry a small amount of sample;

[0009] The protective sleeve is an axially expandable flexible sleeve fitted over the outside of the support rod, and has a fixed end and a free end; the fixed end is connected to a limiting structure on the support rod, and the free end can move axially relative to the support rod to switch between a protective state covering the sample ring and an operational state exposing the sample ring.

[0010] The support rod has a sensor channel extending along its axial direction inside, and the end of the sensor channel leads to the sample carrying ring to guide the detection end of the temperature sensor to the sample measurement point.

[0011] Furthermore, a fixing structure is provided on the inner side of the sample carrying ring for positioning and fixing the detection end of the temperature sensor.

[0012] Furthermore, the fixing structure is a protrusion on the inner wall of the sample carrying ring, and the protrusion is fixed to the detection end of the temperature sensor by an adhesive.

[0013] Furthermore, the support rod has a stepped structure and is integrally formed with the sample carrier ring.

[0014] Furthermore, the sample carrier ring is a micro-nano scale ring structure, with its ring width and thickness both between 0.1 mm and 0.2 mm.

[0015] The protective sleeve is preferably a stretchable corrugated structure, made of low-temperature resistant and lightweight polytetrafluoroethylene (PTFE). One end of the protective sleeve is fixedly connected to the baffle at the middle and rear of the support rod, and the other end, if extended to the outermost edge, can be used to provide effective physical protection and thermal shielding for the sample ring and the sample on it during cooling and reheating, preventing interference from external airflow, thermal radiation, etc., on the heating and cooling process.

[0016] The corrugated structure has good flexibility, which can adapt to the thermal expansion and contraction of materials during cooling or reheating, and can be retracted when not in use to expose the sample loading area for easy operation.

[0017] The support rod is located inside the protective sleeve and extends axially to its outside, providing support. This support rod has a simple structure and high rigidity.

[0018] Preferably, the support rod is made of a low thermal conductivity resin material to limit the heat diffusion path.

[0019] The support rod preferably adopts a stepped structure to ensure the rigidity of the support, which on the one hand improves the connection strength, and on the other hand reduces the local volume and enhances the structural stability.

[0020] Preferably, the portion of the support rod area connected to the sample carrying ring has a minimum thickness of 0.2 mm to minimize the thermal impact on the sample.

[0021] Preferably, the rear half of the support rod has an axial through hole extending into the protective tube, which is used to pass through and install the temperature sensor probe, so as to realize temperature monitoring of the entire process of sample temperature change.

[0022] The sample carrier ring is located at the end of the support rod and has a micro-nano scale ring structure for supporting a fixed trace amount of biological sample. Preferably, the sample carrier ring is fabricated using 3D micro-nano printing technology, which has high dimensional accuracy and mechanical strength, and can be customized according to the sample volume.

[0023] Preferably, the inner diameter of the sample carrying ring is 0.4 to 10 mm, suitable for micro-droplet samples with a volume of 0.2 μL to 10 μL.

[0024] Preferably, in order to improve the heat exchange efficiency between the sample and the cooling source, the width and thickness of the ring are both controlled between 0.1 mm and 0.2 mm to increase the contact area between the sample and the low-temperature cold source.

[0025] Preferably, to enable real-time monitoring of sample temperature, a small protrusion is provided on the sample ring to fix the position of the thermocouple probe and prevent it from shifting or falling off. This protrusion can be bonded to the thermocouple probe using a low-temperature adhesive to ensure that the temperature probe remains in close contact with the sample during freezing and rewarming processes.

[0026] Preferably, the temperature sensor used should have an extremely fine wire diameter to minimize its interference with the sample's thermal conduction and microenvironment structure, thereby ensuring the accuracy of temperature control and the safety of the sample.

[0027] Compared with the prior art, the beneficial effects of this utility model are as follows:

[0028] 1. Strong functional expandability: The support rod has reserved sensor channels to support real-time temperature measurement needs of samples during freezing or rewarming.

[0029] 2. Compact and stable structure: The protective sleeve and support system are designed in a coordinated manner to effectively avoid disturbance to the sample during processing.

[0030] 3. Rapid thermal response: The sample-carrying cryo-arm adopts a micro-nano ring structure to carry the sample, which increases the contact area between the sample and the liquid nitrogen cold source and reduces the influence of the sample-carrying device on the thermodynamic changes of the sample.

[0031] 4. High adaptability: The size of the sample carrying ring can be customized as needed, and it is suitable for various micro-droplet operations. Attached Figure Description

[0032] Figure 1 This is a schematic diagram of a vitrified cooling micro-sample carrier device for extended temperature measurement according to the present invention.

[0033] Figure 2 A schematic diagram of the structure design for scalable temperature measurement;

[0034] In the picture:

[0035] 1-Protective sleeve;

[0036] 2-Support rod, 201-Limiting structure, 202-Middle stepped structure, 203-Rear section, 204-Sensor channel, 205-Front section;

[0037] 3-Sample carrier ring, 301-Fixing structure;

[0038] 4-Temperature sensor. Detailed Implementation

[0039] To make the objectives, technical solutions, and advantages of the embodiments of this utility model clearer, the technical solutions of the embodiments of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this utility model. Based on the embodiments of this utility model, all other embodiments obtained by those skilled in the art without creative effort are within the protection scope of this utility model.

[0040] Please see Figure 1 and Figure 2 As shown in the figure, this utility model provides a glass-cooled micro sample carrier device with extended temperature measurement function, which consists of three parts: a protective sleeve 1, a support rod 2, and a sample carrier ring 3.

[0041] The protective sleeve 1 is connected to the support rod 2 via a limiting structure 201, which is located in the middle and rear part of the support rod. The limiting baffle 201, the support rod 2, and the sample carrier ring 3 are integrally formed.

[0042] The protective sleeve 1 is a corrugated structure with flexible and extensible properties, preferably made of a polymer material with low-temperature toughness. One end of it is fixedly connected to a limiting baffle 201 at the middle and rear of the support rod, achieving stable support through the limiting baffle 201. The protective sleeve can extend to the outermost edge of the sample carrying ring 3, thereby protecting the sample; at the same time, it can also retract axially to fully expose the sample and prevent it from being obstructed during sample processing.

[0043] The support rod 2 is located inside the protective sleeve 1, and is preferably made of a resin material with low thermal conductivity to suppress the axial thermal diffusion effect of the cold source or laser heat source. The support rod 2 extends axially through the protective sleeve 1 to the outside, and its distal end is integrally formed with the sample carrying ring 3 to ensure structural strength and coaxial stability. In addition, the front section of the support rod is designed with a central stepped structure 202 to reduce the overall weight of the rod and improve local deformation resistance.

[0044] A through-hole 204 is provided on one side of the rear section 203, extending into the internal space of the protective sleeve, for embedding a temperature sensor 4 with an extremely fine wire diameter, providing a channel for temperature signal transmission, fixing the temperature sensor and reducing thermal disturbance caused by the movement of the sensor body.

[0045] The sample carrier ring 3 is a closed ring structure at the micro-nano scale, with its entirety located at the front section 205. This extremely fine ring structure avoids large-area contact with the sample, increasing the contact area between the sample and the liquid nitrogen cold source. It is preferably manufactured using 3D micro-nano printing technology. Through hydrophilic surface treatment, this ring structure improves the adhesion and stability of micro-droplets (0.2–10 μL), ensuring the sample's stability during vitrification cooling and temperature measurement.

[0046] A small protrusion, or fixing structure 301, is provided on the inner side of the sample carrying ring 3 to fix the position of the thermocouple probe and prevent it from shifting or falling off. This protrusion can be bonded to the thermocouple probe with a low-temperature special adhesive to ensure that the temperature measuring probe is in close contact with the sample during the measurement process, thus ensuring the accuracy and reliability of the temperature measurement point.

[0047] The recommended structural parameters for the sample carrier ring in this embodiment are as follows: the inner diameter of the sample carrier ring 3 is 0.4-10 mm, and the ring width and thickness are both 0.1-0.2 mm, to accommodate the cooling requirements of samples of different volume levels. The minimum thickness of the support rod's front end is preferably 0.2 mm. The small protrusion on the inner side of the sample carrier ring is preferably 0.1 mm * 0.1 mm * 0.05 mm in size. This structure is used to relatively fix the position of the thermocouple temperature probe and is bonded using a low-temperature adaptable adhesive. In addition, if necessary, the protrusion can also shield the temperature probe from direct irradiation by external physical fields such as lasers, preventing errors in temperature measurement data.

[0048] The device possesses excellent thermal response characteristics, mechanical strength, and structural adaptability, making it suitable for use as a vitrification cooling experimental platform under extreme low-temperature conditions. It is particularly suitable for the freezing and rewarming processes of micro-biological samples such as cells and tissue droplets that require precise temperature monitoring.

[0049] Example 1

[0050] Prepare an aqueous solution of 2.5 mol / L ethylene glycol, 2.5 mol / L propylene glycol, and 0.5 mol / L trehalose as a cryoprotectant at an initial temperature of 25°C. Ensure the protective sleeve is in its initial contracted state before placing the sample. Use a pipette to select 1 μL of this solution as the sample to be cooled. Place the sample on a sample carrier with an inner diameter of 1 mm and a width and thickness of 0.1 mm. After sample placement, extend the protective sleeve to protect the internal support rod and the sample. Then, immerse the sample carrier in liquid nitrogen to complete the rapid cooling process.

[0051] Example 2

[0052] When monitoring the temperature change of the sample during the cooling process, first select a thermocouple with a wire diameter of 50 micrometers and pass it through the axial through-hole. Fix the thermocouple probe on the protrusion on the inner side of the sample carrier ring. Confirm that the protective sleeve is in the contracted state. Prepare an aqueous solution of 2.5 mol / L ethylene glycol, 2.5 mol / L propylene glycol, and 0.5 mol / L trehalose as a cryoprotectant with an initial temperature of 25°C. Confirm that the protective sleeve is in the initial contracted state before placing the sample. Use a pipette to select 1 μL of this solution as the sample to be cooled. Place the sample on the sample carrier device with an inner diameter of 1 mm and a ring width and thickness of 0.1 mm. After the sample is placed, the protective sleeve extends to protect the internal support rod and the sample. Then immerse the sample carrier device in liquid nitrogen to complete the rapid cooling process. After 1.5 seconds, the sample temperature drops to 77 K (liquid nitrogen temperature). By connecting an external data acquisition device with a temperature sensor, the cooling rate of the sample can be measured to be up to 8800 K / min.

[0053] Example 3

[0054] When monitoring the temperature change of a sample during the rewarming process in a 37°C water bath is required, a temperature sensor must be embedded during the cooling phase (see Example 2). After removing the sample and transferring it to the 37°C water bath for rewarming, the temperature sensor connected to an external data acquisition device can measure the heating rate of the sample in Example 2, which can reach 10000 K / min. Once the sample has warmed to room temperature, the protective sleeve is removed, and the sample is processed.

[0055] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this utility model, and not to limit it. Although this utility model has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this utility model.

Claims

1. A vitrified cooling microsample carrier device for extended temperature measurement, characterized in that, include: The support rod (2) is in the shape of an inverted step and is used to prevent liquid nitrogen from escaping. A sample carrying ring (3) is disposed at the end of the support rod (2) and is used to carry a small amount of sample; The protective sleeve (1) is an axially expandable flexible sleeve, which is sleeved on the outside of the support rod (2) and has a fixed end and a free end; the fixed end is connected to the limiting structure (201) on the support rod (2), and the free end can move axially relative to the support rod (2) to switch between a protective state covering the sample ring (3) and an operational state exposing the sample ring (3).

2. The extended temperature measurement vitrification cooling microsample carrier device according to claim 1, characterized in that, The support rod (2) has a sensor channel (204) extending along its axial direction inside. The end of the sensor channel (204) leads to the sample ring (3) and is used to guide the detection end of the temperature sensor (4) to the sample measurement point.

3. The extended temperature measurement vitrification cooling microsample carrier device according to claim 2, characterized in that, The inner side of the sample carrier ring (3) is provided with a fixing structure (301) for positioning and fixing the detection end of the temperature sensor (4).

4. The extended temperature measurement vitrification cooling microsample carrier device according to claim 3, characterized in that, The fixing structure (301) is a protrusion protruding from the inner wall of the sample carrier ring (3), and the protrusion is fixed to the detection end of the temperature sensor (4) by an adhesive.

5. The extended temperature measurement vitrification cooling microsample carrier device according to claim 1, characterized in that, The support rod (2) has a stepped structure and is integrally formed with the sample carrier ring (3).

6. The extended temperature measurement vitrification cooling microsample carrier device according to claim 1, characterized in that, The sample carrier ring (3) is a micro-nano scale ring structure with a ring width and thickness between 0.1 mm and 0.2 mm.

7. The extended temperature measurement vitrification cooling microsample carrier device according to claim 6, characterized in that, The surface of the sample carrier ring (3) is treated with a hydrophilic coating.

8. The extended temperature measurement vitrification cooling microsample carrier device according to claim 1, characterized in that, The protective sleeve (1) has a rigid corrugated structure and flexible extensibility to adapt to thermal expansion and contraction during cooling or reheating.

9. The extended temperature measurement vitrification cooling microsample carrier device according to claim 1, characterized in that, The sample carrier ring is manufactured using 3D micro-nano printing technology. Its size can be customized according to the volume of the sample to be cooled. The inner diameter of the ring is 0.4 to 10 mm, and the ring width and thickness are both set to 0.1 to 0.2 mm.

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