CN1140829C - Heat sink type cryogenic biological sample platform for scanning probe microscope - Google Patents
Heat sink type cryogenic biological sample platform for scanning probe microscope Download PDFInfo
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- CN1140829C CN1140829C CNB001301225A CN00130122A CN1140829C CN 1140829 C CN1140829 C CN 1140829C CN B001301225 A CNB001301225 A CN B001301225A CN 00130122 A CN00130122 A CN 00130122A CN 1140829 C CN1140829 C CN 1140829C
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Abstract
The present invention relates to a heat sink type cryogenic biological sample platform which is composed of a vacuum imaging cavity and a narrownecked specimen platform, wherein the narrownecked specimen platform is arranged in the vacuum imaging cavity. The platform surface of a temperature sensor is fixedly arranged on the top end of the narrow neck of the specimen platform, a molecular sieve is arranged in a slot of a large platform surface on the lower part of the specimen platform, and the external surface of the specimen platform is covered with an insulating layer. The narrow neck is exposed outside the cavity, and a low expansion alloy thin-wall pipe is sheathed around the narrow neck. The upper end and the lower end of the alloy thin-wall pipe are respectively welded on the platform surface and a cover plate of the vacuum imaging cavity. The heat sink type cryogenic biological sample platform has the advantages of simple structure and low price and can be matched with a common scanning probe microscope under the condition of not increasing large and expensive auxiliary equipment. The function of the heat sink type cryogenic biological sample platform extends from condensation point temperature to low temperature.
Description
Invention field
The present invention relates to a kind of biological sample platform, thereby be particularly related to a kind of initial temperature can preset can keep the nano grade biological sample below certain set point temperature can with the matching used heat sink type cryogenic biological sample platform that is used for scanning probe microscopy of nearly all scanning probe microscopy, can make plain scan probe microscope function expand to scope from the set point temperature to lower temperature, also can be used as general cryogenics observation platform, and simple in structure, cheap.
Background technology
Life process occurs in three space scales: the nanoscale relevant with the organic molecule behavior, the micro-meter scale relevant with the interior individual cells behavior of tissue reach the macro-scale relevant with whole living organism behavior.The low temperature that surpasses the freezing temperature of water is embodied in nanometer (molecule) and micron (cell) yardstick simultaneously to the influence of biosystem.Phase transformation meeting in the cell membrane lipid bilayer influences the function of inside and outside solution of its isolated cell and control of quality transmission, thereby to understand and control this phase transition process be one of of paramount importance field in the research of microscale bio-heat transfer, and development advanced person's measuring technique will greatly be promoted the understanding to related mechanism in this problem.People adopt various approach such as methods such as low temperature scanning Electronic Speculum, X-ray diffraction and mathematical simulation that heat transfer, mass transfer and a series of physics that causes thus, the chemical behavior that occur in inside and outside the cell are studied for a long time, but the understanding to this mechanism is still serious deficient so far, and one of reason is that very difficult pair cell and inner structure thereof are carried out Direct observation.Along with progress of modern technology, the human new tool of more and more having obtained more more perfect observation observation of minute biological samples.At present, can think that scanning probe microscopy (general names of atomic force microscope, scanning tunnel microscope etc.) is best suited for one of instrument of observing in pair cell on molecular level.
The application of scanning probe microscopy in biomedical research just constantly obtains expanding, this Direct observation about information on the cellular level is very crucial for the announcement of biological phenomena, although the micromechanism of many biological samples can be observed by the way of electronics or optical microscope, but most of resolution are only in micron dimension, thereby the atom of Direct observation biological sample and molecular structure become the problem that is rich in temptation for a long time, and the invention of scanning probe microscopy has promoted this process greatly.But some present high precision images mainly obtain on big molecule, and the precision and the repdocutbility of different samples are relatively poor, this be because, moisture big molecule is a kind of flexible material under the room temperature on the one hand, can deform because of the operation of probe even destroy, therefore the gained precision of images is lower and repeated relatively poor, and the water vapor of biological sample has also increased the difficulty that signal is differentiated; On the other hand, the feasible very difficult high precision imaging that realizes of macromolecular thermal motion under the room temperature.The measure that solves above-mentioned difficulties is directly to observe biological sample at low temperatures, the scanning probe microscopy of Jian Liing is called the low temperature atomic force microscope thus, this is because at low temperatures, molecular thermalmotion is greatly suppressed, thus the pressure release surface of biological sample can hardening can guarantee to obtain than room temperature under higher resolution.Shao and co-worker thereof " the biological low temperature atomic force microscope of using: brief review " (the 66th volume 141-152 page or leaf in U.S.'s " ultramicroscope ", 1996) (Shao Z.andY.Zhang, Biological cryo atomic force microscopy:a brief review, Ultramicroscopy, vol.66, pp.141-152,1996) designed a kind of low temperature atomic force microscope.Different with the common atomic force microscope of operating under atmospheric environment is, in this system, the micro-head of the atomic force of being made up of laser diode, photodetector and scanner is placed in one and includes in the low temperature environment of liquid nitrogen, the Dewar flask splendid attire liquid nitrogen that this low temperature environment employing internal diameter is 28in is realized, nitrogen vapor pressure in the Dewar flask can pass through valve regulated, and a little more than external atmosphere pressure, the vibration that causes with the generation of avoiding owing to the liquid nitrogen bubble; The imaging chamber of atomic force microscope is positioned at about 1-2in place on the liquid nitrogen surface, and the temperature of imaging sample depends on that just the distance of atomic force microscope and liquid nitrogen is far and near, and the low temperature atomic force microscope of Shao etc. can be implemented in the measurement of 77-220K scope.In obtaining the process of image, imaging chamber must keep sealing, to eliminate the disturbance that brings owing to the nitrogen vapor circulation.Many new biomedical research fields have been opened up in the realization of low temperature atomic force microscope, and it has been used to the observation of a series of superfine microbiological specimens such as immunoglobulin (Ig), virus, DNA, red blood cell, cell membrane etc., and its scope just constantly obtains expanding.But the device of Shao etc. is structurally very complicated, and it is bigger to implement difficulty, and expensive, and the liquid nitrogen consumption is very big, and it is more to take up space, and experiment expends many.
Germany Omicron company is from another kind of structure (Hou Shimin etc., alternating temperature ultrahigh vacuum scanning probe microscopy, Modern Scientific Instruments, No.3, pp.19-21,2000), developed so-called alternating temperature ultrahigh vacuum scanning probe microscopy, can be to the sample imaging between 25K-1100K.This system adopts the liquid helium continuous stream to cross certain heat interchanger and realizes, heat interchanger links to each other with specimen holder by the copper pigtail line, and utilizes heat conduction to reduce sample temperature.Under the mechanical pump effect, liquid helium is sucked out from Dewar flask, and Dewar flask enters liquid helium continuous flow refrigeratory and heat interchanger carries out heat interchange by infusing.The heated sample frame also can make sample change to 1100K from room temperature.This device also exists complex structure and higher-priced problem.
Summary of the invention
It is a kind of simple in structure that the object of the invention is to provide, cheap cryogenic biological sample platform, thereby particularly a kind of initial temperature can preset can keep the nano grade biological sample below certain set point temperature can with the matching used heat sink type cryogenic biological sample platform of nearly all scanning probe microscopy, can be in that not increase the plain scan probe microscope necessary huge, under the situation of expensive utility appliance, make the function of plain scan probe microscope expand to scope from the set point temperature to lower temperature, simultaneously, this cryogenic biological sample platform also can be used as general cryogenics observation platform.
Embodiment of the present invention are as follows:
The heat sink type cryogenic biological sample platform that is used for scanning probe microscopy provided by the invention: comprise heat sink A and vacuum (-tight) housing 9 two parts that link to each other with vacuum pump 10, heat sink A places on the scanning probe microscopy platform 15, places among the vacuum (-tight) housing 9 again; Described heat sink A is made up of the solid slim neck sample stage 13 and the vacuum imaging chamber 6 of big specific heat capacity material, the thin neck 14 tops fixed installation one of solid slim neck sample stage 13 is in order to place the table top 1 of cryogenic biological sample, mounting temperature sensor 11 on the table top 1, in the groove on the big table top in bottom of solid slim neck sample stage 13 molecular sieve 4 is housed, cover heat insulation layer 5 on the big table top outside surface in the bottom of solid slim neck sample stage 13, vacuum imaging chamber 6 is a hollow cylinder by the low-expansion alloy making, solid slim neck sample stage 13 places the inner chamber in vacuum imaging chamber 6, its thin neck 14 exposes outside the vacuum imaging chamber 6, suit low-expansion alloy thin walled tube 2 around the thin neck 14, on the low-expansion alloy thin walled tube 2, the lower end is welded on respectively on the cover plate 3 in the table top 1 of solid slim neck sample stage 13 and vacuum imaging chamber 6, the cover plate 3 in vacuum imaging chamber 6 is provided with vacuum orifice 8, vacuumizes between solid slim neck sample stage 13 and the vacuum imaging chamber 6; For guaranteeing that cold leaks to the platform of scanning probe microscopy, vacuum imaging chamber 6 is fixed in the adiabatic substrate 12.
Solid slim neck sample stage 13 is made by solid big specific heat capacity material such as copper billet, and make by the less Invar alloy of thermal expansivity in low-expansion alloy thin walled tube 2 and vacuum imaging chamber 6.Gap between Invar alloy and copper billet is evacuated, because thin neck sample stage is provided with molecular sieve 4, the back of can guaranteeing to lower the temperature obtains high-quality vacuum environment, thereby reaches good effect of heat insulation.Heat sink the certain position of A is provided with thermopair 15 to monitor actual heat sink temperature level.Heat sink A very light in weight generally between 50g-150g, is placed in the vacuum (-tight) housing 9, and vacuum (-tight) housing 9 can be a cloche, and connects the vacuum pump 10 vacuumize, reducing the heat radiation of sample surfaces, and eliminates reading of the presumable influence of moisture image of sample surfaces as far as possible.Because the sample stage thermal capacitance is very big, and treat that the observing samples volume is minimum, thereby heat sink type example platform temperature can keep relative stablizing, satisfy the observation of scanning probe microscopy under low temperature necessarily thus.
For guaranteeing that cold leaks to the scanning probe microscopy platform, the present invention further places vacuum imaging chamber 6 bases in the one adiabatic substrate 12, thereby has realized above-mentioned target well.According to Testing requirement this heat sink type example platform is preset in certain hour in certain low-temperature receiver such as the Dewar bottle, watches it and reach stable, taking-up places on the scanning probe microscopy platform, and sample is put, and can observe.After also sample stage can being placed certain hour in air, set aside to again on the scanning probe microscopy platform, can realize the sample observation under the higher a little temperature conditions thus.Like this, the low-temperature biological material that can satisfy under the different temperatures is observed.
The present invention is simple in structure, price is very cheap, and performance meets the demands fully, and the function that basic and complex structure, expensive low temperature scanning probe microscope platform are in the past realized is suitable, and its biggest advantage is can be directly and the supporting use of existing scanning probe microscopy.
Weight of the present invention is less, generally between 50g-150g, and the unlikely driving function that influences the scanning probe microscopy piezoelectric crystal.And because copper billet welding is suspended in the exocoel, even because alternating temperature itself can deform, but this deformation direction down, thereby any change still almost do not take place in sample surfaces, thereby guarantees the imaging of scanning probe microscopy.Can think that this is one of most important design proposal of the present invention.Because the copper billet thermal capacitance is bigger, and is evacuated in the imaging chamber, cold is difficult for scattering and disappearing on the sample stage, thus can guarantee sample keep within the specific limits constant, thereby satisfy the observation of scanning probe microscopy.Under different low temperature level observation material especially the pattern of nano grade biological material be the direction of exploring, the present invention can help application in this respect.
The heat sink type cryogenic example platform and the peripheral unit thereof that are used for the scanning probe microscopy of nanoscale low temperature cytological observation, thereby be particularly related to a kind of initial temperature can preset can keep observation of minute biological samples below certain set point temperature (from 0 ° of C to liquid nitrogen temperature-196 ℃ or lower) can with the matching used example platform of nearly all scanning probe microscopy, its role is to common scanning probe microscopy function is expanded to scope from the set point temperature to lower temperature, can be by the example platform temperature being preset the temperature that changes sample on the platform on one's own initiative, and the specimen material surface that has changed temperature carried out imaging, because the heat sink type example platform that is adopted is simple in structure, can with the supporting use of nearly all scanning probe microscopy, extent of alternating temperature is wide, need not to change the structure of existing scanning probe microscopy, and price is very cheap, can satisfy the requirement of biological sample observation under the different temperatures preferably, this is very favourable for promoting the use of of instrument.The most significant characteristics of native system are to introduce the notion of heat sink type cryogenic example platform, thereby guarantee the relative independentability of example platform, and realize the observation of material in the different temperatures scope.Low temperature scanning probe microscope is to attempt the target explored in the nanosecond science and technology field, and this example platform and the method that is adopted thereof have had not yet to see report.
Description of drawings
Fig. 1 is a structural representation of the present invention;
Fig. 2 is the structural representation of heat sink A;
Wherein: solid slim neck sample stage 1 low-expansion alloy thin walled tube 2
Vacuum (-tight) housing 9 vacuum pumps 10
Solid slim neck sample stage 13 thin necks 14
Scan-probe platform 15
Embodiment
Further describe the present invention below in conjunction with the drawings and specific embodiments:
Known the heat sink type cryogenic biological sample platform that is used for scanning probe microscopy provided by the invention by figure: comprise heat sink A and vacuum (-tight) housing 9 two parts that link to each other with vacuum pump 10, heat sink A places among the vacuum (-tight) housing 9; Described heat sink A is made up of the solid slim neck sample stage 13 and the vacuum imaging chamber 6 of big specific heat capacity material, the thin neck 14 tops fixed installation one of solid slim neck sample stage 13 is in order to place the table top 1 of cryogenic biological sample, mounting temperature sensor 11 on the table top 1, in the groove on the big table top in bottom of solid slim neck sample stage 13 molecular sieve 4 is housed, be coated with heat insulation layer 5 on the outside surface on the big table top in bottom of solid slim neck sample stage 13, vacuum imaging chamber 6 is a hollow cylinder by the low-expansion alloy making, solid slim neck sample stage 13 places the inner chamber in vacuum imaging chamber 6, the thin neck of solid slim neck sample stage 13 exposes vacuum imaging chamber 6, suit low-expansion alloy thin walled tube 2 around it, on the low-expansion alloy thin walled tube 2, the lower end is welded on respectively on the cover plate 3 in the table top 1 of solid slim neck sample stage 13 and vacuum imaging chamber 6, the cover plate 3 in vacuum imaging chamber 6 is provided with vacuum orifice 8, vacuumizes between solid slim neck sample stage 13 and the vacuum imaging chamber 6; For guaranteeing that cold leaks to the platform of scanning probe microscopy, vacuum imaging chamber 6 be fixed on adiabatic substrate 12 on.
Solid slim neck sample stage 13 is made by solid big specific heat capacity material such as copper billet, and make by the less Invar alloy of thermal expansivity in low-expansion alloy thin walled tube 2 and vacuum imaging chamber 6.Gap between Invar alloy and copper billet is evacuated, because thin neck sample stage is provided with molecular sieve 4, the back of can guaranteeing to lower the temperature obtains high-quality vacuum environment, thereby reaches good effect of heat insulation.Heat sink the certain position of A is provided with thermopair 11 to monitor actual heat sink temperature level.Heat sink A is placed in the vacuum (-tight) housing 9, and vacuum (-tight) housing 9 can be a cloche, and connects the vacuum pump 10 vacuumize, reducing the heat radiation of sample surfaces, and eliminates reading of the presumable influence of moisture image of sample surfaces as far as possible.Because the sample stage thermal capacitance is very big, and treat that the observing samples volume is minimum, thereby heat sink type example platform temperature can keep relative stablizing, satisfy the observation of scanning probe microscopy under low temperature necessarily thus.
Number of assembling steps of the present invention: after in the groove on the big table top in the bottom of solid slim neck sample stage 13 molecular sieve 4 being set, will be coated with formation heat insulation layer 5 with thermal insulation material such as glass fibre around it; Table top 1 with the end of the thin neck 14 of solid slim neck sample stage 13 welds together with the low-expansion alloy thin walled tube 2 that low bulk Invar alloy constitutes again, with cover plate 3 welding in low-expansion alloy thin walled tube 2 bottoms and vacuum imaging chamber 6, weld with base 7 in vacuum imaging chamber 6 afterwards; Whole welding process will be guaranteed along circumferential seal.Simultaneously, vacuum imaging chamber 6 and base 7 welding constitute heat sink type A, and will be vacuumized between solid slim neck sample stage 13 and the vacuum imaging chamber 6 by vacuum orifice;
The very light in weight of heat sink A generally between 50g-150g, and is inserted in the vacuum (-tight) housing 9, and vacuum (-tight) housing 9 is vacuumized by vacuum pump 10.
From the above mentioned, the low temperature example platform that the present invention adopts is simple in structure, good stability, it is cheap that price then is tending towards, make relatively easy, data acquisition and handle very convenient, no complicated circuit, the hot state ratio of thermometric and assess sample is easier to, extent of alternating temperature is wide.The research of material character and the observation of nanotopography under the suitable different low temperature, so carry out the test of relevant thermophysical property.
During use, cryogenic biological sample platform of the present invention is placed low-temperature receiver in advance, after treating its cooling and reaching evenly, taking-up places on the raw sample platform 15 of plain scan probe microscope, cryogenic biological sample is held on the table top 1 of this sample stage, and vacuum imaging chamber 6 vacuumized, can begin the imaging of scanning probe microscopy afterwards.If need to change sample temperature, realize after sample stage can being shelved certain hour in air, thereby reach slightly higher new sample temperature.Thus, promptly realize the imaging of the specimen material pattern under the different low temperature.And can study the influence of cooling, and measure relevant important hot rerum natura to material pattern and relevant physical property.
Claims (7)
1. heat sink type cryogenic biological sample platform that is used for scanning probe microscopy, it is characterized in that: this cryogenic biological sample platform comprises heat sink (A) and vacuum (-tight) housing (9) two parts that link to each other with vacuum pump (10), and heat sink (A) places among the vacuum (-tight) housing (9); Described heat sink (A) is made up of the solid slim neck sample stage (13) and the vacuum imaging chamber (6) of big specific heat capacity material, thin neck (14) the top fixed installation one of solid slim neck sample stage (13) is in order to place the table top (1) of cryogenic biological sample, table top (1) is gone up mounting temperature sensor (11), in the groove on the big table top in bottom of solid slim neck sample stage (13) molecular sieve (4) is housed, be coated with heat insulation layer (5) on the outside surface on the big table top in bottom of solid slim neck sample stage (13), vacuum imaging chamber (6) is a hollow cylinder by the low-expansion alloy making, solid slim neck sample stage (13) places the inner chamber in vacuum imaging chamber (6), the thin neck of solid slim neck sample stage (13) exposes vacuum imaging chamber (6), suit low-expansion alloy thin walled tube (2) around it, on the low-expansion alloy thin walled tube (2), the lower end is welded on respectively on the cover plate (3) in the table top (1) of solid slim neck sample stage (13) and vacuum imaging chamber (6), the cover plate (3) in vacuum imaging chamber (6) is provided with vacuum orifice (8), vacuumizes between solid slim neck sample stage (13) and the vacuum imaging chamber (6).
2. by the described heat sink type cryogenic biological sample platform that is used for scanning probe microscopy of claim 1, it is characterized in that: solid slim neck sample stage (13) is made of copper.
3. by the described heat sink type cryogenic biological sample platform that is used for scanning probe microscopy of claim 1, it is characterized in that: vacuum imaging chamber (6) is made by the Invar alloy.
4. by the described heat sink type cryogenic biological sample platform that is used for scanning probe microscopy of claim 1, it is characterized in that: temperature sensor (11) is a thermocouple temperature sensor.
5. by the described heat sink type cryogenic biological sample platform that is used for scanning probe microscopy of claim 1, it is characterized in that: vacuum (-tight) housing (9) is a cloche.
6. by claim 1,2,3, the 4 or 5 described heat sink type cryogenic biological sample platforms that are used for scanning probe microscopy, it is characterized in that: the base (7) in vacuum imaging chamber (6) is fixed in the adiabatic substrate (12).
7. by claim 1,2,3, the 4 or 5 described heat sink type cryogenic biological sample platforms that are used for scanning probe microscopy, it is characterized in that: the weight of heat sink (A) is between 50g-150g.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB001301225A CN1140829C (en) | 2000-09-29 | 2000-09-29 | Heat sink type cryogenic biological sample platform for scanning probe microscope |
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| CNB001301225A CN1140829C (en) | 2000-09-29 | 2000-09-29 | Heat sink type cryogenic biological sample platform for scanning probe microscope |
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| CN1140829C true CN1140829C (en) | 2004-03-03 |
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1558424B (en) * | 2004-01-16 | 2010-04-07 | 中国科学院长春应用化学研究所 | Gold thin film substrate making method for scanning probe microscope |
| CN100495036C (en) * | 2004-04-27 | 2009-06-03 | 中国科学院理化技术研究所 | Chip type measuring device for rapidly determining optimum low-temperature storage parameters of biological samples |
| CN101865729B (en) * | 2009-04-15 | 2012-03-07 | 中国科学院半导体研究所 | Method for measuring cavity surface temperature of semiconductor laser |
| KR102328101B1 (en) * | 2015-07-07 | 2021-11-17 | 삼성전자주식회사 | Probe card, thermal insulation cover assembly for probe card, and apparatus of testing semiconducotr device including the probe card |
| CN107389455B (en) * | 2017-09-05 | 2023-06-06 | 中国工程物理研究院流体物理研究所 | Cooling device and method for initial temperature of sample in magnetic drive oblique wave compression |
| CN107677771B (en) * | 2017-09-15 | 2019-12-13 | 北京大学 | Ultrahigh vacuum low-temperature sample table |
| CN112630247A (en) * | 2020-12-31 | 2021-04-09 | 厦门超新芯科技有限公司 | Frozen sample platform of scanning electron microscope |
| CN117739612B (en) * | 2023-07-12 | 2024-07-09 | 西湖大学 | Liquid helium consumption free circulating refrigerating system and liquid helium temperature zone electron microscope |
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