KR101879343B1 - Multi-purpose micro-calorimeter for bio sample and calorific value measuring method for bio sample using it - Google Patents

Abstract

The present invention relates to a multipurpose micro-calorimeter for biosamples capable of simultaneously measuring changes in calorific value and enthalpy physical properties generated in a small micro sample of micro liters (μl), and a method for manufacturing the same.
The multi-purpose micro-calorimeter for a bio sample according to the present invention comprises a substrate part polished on one side or both sides of a wafer, an insulating layer deposited on the substrate part and composed of two or more different compound layers, And a protective layer formed on the insulating layer to cover the insulating layer, the thermopile, and the heater, wherein the insulating layer is formed on the upper surface of the insulating layer, The heater is formed to alternately heat the bio sample to be placed on the protective layer by applying an alternating current.

Description

TECHNICAL FIELD [0001] The present invention relates to a multi-purpose micro-calorimeter for biosamples, and a method of manufacturing the same. [0002] MULTI-

The present invention relates to a multi-purpose micro-calorimeter for a biosample and a method of manufacturing the micro-calorimeter, and more particularly, to a multi-purpose micro-calorimeter for a bio sample, And a method for producing the same.

The population and total calorific value are measured separately when culturing general bacteria. That is, for example, the number of existing microorganisms is measured by the plate counting method or the optical density method, while the heating value is measured at the same time using a separate calorimeter.

However, since the growth of microorganisms changes drastically depending on culture conditions, it is difficult to measure the exact calorific value of a single individual when the population and caloric value can not be measured simultaneously.

Korean Patent No. 10-0911090 is a technology for measuring the specific heat / heat capacity of a bio sample. The prior art uses a curve fitting of the temperature response curve after heating to measure the specific heat / heat capacity of the sample, so that there is a problem that a lot of temperature data is required to obtain an accurate measurement value.

Korean Registered Patent No. 10-0911090: Improved Accuracy of a Micro Calorimeter Device Korean Patent No. 10-1050517: Micro-heat flux sensor and manufacturing method thereof

SUMMARY OF THE INVENTION The present invention has been made in order to solve the above problems, and it is an object of the present invention to provide a multi-purpose micro-calorimeter for biosamples and a method for manufacturing the same, which can measure a change in heat capacity over a long period of time in real- have.

The multi-purpose micro-calorimeter for a bio sample according to the present invention comprises a substrate part polished on one side or both sides of a wafer, an insulating layer deposited on the substrate part and composed of two or more different compound layers, And a protective layer formed on the insulating layer to cover the insulating layer, the thermopile, and the heater, wherein the insulating layer is formed on the upper surface of the insulating layer, The heater is formed to alternately heat the bio sample to be placed on the protective layer by applying an alternating current.

Wherein the insulating layer includes a first insulating portion formed by depositing silicon dioxide on an upper surface of the substrate portion, and a second insulating portion formed by depositing silicon nitride on the first insulating portion, wherein the first insulating portion has a thickness of 0.7 to 0.9 mu m And the second insulating portion is formed to have a thickness of 0.2 to 0.4 탆.

Wherein the thermopile includes a gold coating layer and a chromium coating layer, wherein the gold coating layer is spaced apart by a predetermined distance, both ends of the chromium coating layer are laminated on the gold coating layer, .

It is preferable that the gold coating layer has a thickness of 0.1 to 0.3 탆 and the chromium coating layer has a thickness of 0.2 to 0.4 탆.

It is preferable that the protective layer is formed by depositing silicon dioxide, and the thickness from the insulating layer to the top of the protective layer is 0.9 to 1.1 탆.

Preferably, the substrate portion includes a cavity formed by etching the insulating layer, the thermopile, and the opposed surface opposed to the one surface on which the heater is formed to concentrate heat flow from the heater into the thermopile.

A method of manufacturing a multi-purpose micro-calorimeter for a biosample according to the present invention includes the steps of forming a polished substrate portion on one or both surfaces of a wafer formed of silicon, depositing an insulating layer on at least one surface of the substrate portion, The method comprising the steps of: forming two thermocouples spaced apart from one another and spaced apart thermopiles on the insulating layer, the thermocouples forming a heater for applying alternating current to the thermopile and the heater, To form a protective layer formed on the substrate.

And forming a cavity by etching the substrate portion on the other side of the substrate portion opposite to the one surface of the substrate portion on which the thermopile and the heater are formed.

Depositing the insulating layer includes depositing silicon dioxide on the substrate portion to form a first insulating portion and depositing silicon nitride on the first insulating portion to form a second insulating portion, The insulating portion and the second insulating portion may be deposited using a chemical vapor deposition method using thermal energy of silicon dioxide and silicon nitride, respectively, wherein the first insulating portion has a thickness of 0.7 to 0.9 mu m, The portion is preferably deposited to have a thickness of 0.2 to 0.4 mu m.

Wherein the step of forming the thermopile comprises forming a gold coating layer on the insulation layer at a predetermined interval and forming a chromium coating layer on both ends of the gold coating layer, A step of continuously forming a first photoresist layer of a (S) pattern on the insulating layer in one direction, depositing gold on the first photoresist layer to form a gold coating layer, Removing the gold coating layer on the insulating layer by using a cleaning agent and continuously forming a second photoresist layer on the insulating layer to connect the gold coatings spaced apart from each other, Depositing chromium on the layer to form a chromium coating layer; and removing the second photoresist layer using a cleaning agent Can.

It is preferable that the heater is formed by depositing chromium so as to be positioned between the thermopiles when chromium is deposited on the second photoresist layer to form a chromium coating layer.

The multi-purpose micrometer calorimeter for a bio sample of the present invention can measure a change in heat capacity over a long period of time in real time using the amplitude of temperature response by AC heating, and at the same time can measure the calorific value and the heat absorption amount generated in the bio sample, The state change can be accurately measured.

1 is a cross-sectional view of one embodiment of a multipurpose micro-calorimeter for a bio sample according to the present invention,
FIG. 2 is a conceptual diagram schematically depicting a production process of the multi-purpose micro-calorimeter for biosample shown in FIG. 1;
FIG. 3 is a conceptual view schematically showing a state in which a bio sample is placed on a multipurpose micro-calorimeter for biosample of FIG. 1 and a calorific value is measured;
4 is a graph showing an input current input to the heater,
FIG. 5 is a graph showing signals sensed in the thermopile after the input current of FIG. 4 is input to the heater,
FIG. 6 is a graph showing the amplitude of the DC component and the AC component by frequency analysis of the sensing signal of FIG. 5,
FIG. 7 is a graph showing an E. coli growth curve calculated by the optical density method,
FIG. 8 is a graph showing the AC signal measured when the E. coli was cultured through the multi-purpose micro calorimeter for the bio sample of the present invention,
9 is a graph of a DC signal measured when the E. coli was cultured through the multi-purpose micro calorimeter for the biosample of the present invention.

Hereinafter, a multi-purpose micro-calorimeter for a bio sample according to an embodiment of the present invention and a method of manufacturing the same will be described in detail with reference to the accompanying drawings. The present invention is capable of various modifications and various forms, and specific embodiments are illustrated in the drawings and described in detail in the text. It is to be understood, however, that the invention is not intended to be limited to the particular forms disclosed, but on the contrary, is intended to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention. Like reference numerals are used for like elements in describing each drawing. In the accompanying drawings, the dimensions of the structures are enlarged to illustrate the present invention in order to clarify the present invention.

The terms first, second, etc. may be used to describe various components, but the components should not be limited by the terms. The terms are used only for the purpose of distinguishing one component from another. For example, without departing from the scope of the present invention, the first component may be referred to as a second component, and similarly, the second component may also be referred to as a first component.

The terminology used in this application is used only to describe a specific embodiment and is not intended to limit the invention. The singular expressions include plural expressions unless the context clearly dictates otherwise. In this application, the terms "comprises", "having", and the like are used to specify that a feature, a number, a step, an operation, an element, a part or a combination thereof is described in the specification, But do not preclude the presence or addition of one or more other features, integers, steps, operations, components, parts, or combinations thereof.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries are to be interpreted as having a meaning consistent with the contextual meaning of the related art and are to be interpreted as either ideal or overly formal in the sense of the present application Do not.

1, a multi-purpose micro-calorimeter 1 for a bio sample according to the present invention comprises a substrate 10 processed with a silicon wafer, an insulating layer 20 deposited on the substrate 10, an insulating layer 20 And a protective layer 50 covering the thermopile 30 and the heater 40, the thermopile 30 and the heater 40 deposited on the upper surface of the thermopile 30 and the heater 40.

The substrate 10 may be polished on both sides of the silicon wafer and polished only on one side of the thermopile 30 and the heater 40 described below. The substrate portion 10 is formed to have a thickness of 490 to 510 탆.

The insulating layer 20 is formed on the substrate 10 by being deposited on both sides of the substrate 10 or only on one side where the thermopile 30 and the heater 40 and the protective layer 50, A layer 20 may be formed.

The insulating layer 20 includes a first insulating portion 21 formed by depositing silicon dioxide and a second insulating portion 22 formed by depositing silicon nitride on the first insulating portion 21. The insulating layer 20 is composed of the first insulating portion 21 and the second insulating portion 22 on which silicon dioxide and silicon nitride, which are mutually different materials, are deposited, and the insulating effect is increased.

The first insulation part 21 and the second insulation part 22 can be deposited using a chemical vapor deposition method. The LPCVD (Low Pressure Chemical Vapor Deposition) method, which is a chemical vapor deposition method using thermal energy, Method, a plasma enhanced chemical vapor deposition (PECVD) method using plasma energy may be applied.

It is preferable that the first insulation portion 21 has a thickness of 0.7 to 0.9 탆 and the second insulation portion 22 has a thickness of 0.2 to 0.4 탆.

The thermopiles 30 are deposited on the insulating layer 20 and are formed on the both sides of the heater 40. The gold coating layer 31 and the chromium coating layer 32 alternate with each other Deposited. After the formation of the insulating layer 20, gold and chromium are deposited using photoresist, respectively. The gold coating layer 31 is deposited so as to be spaced apart from each other by a predetermined distance as shown in a cross-sectional view of FIG. 1 by using a photoresist in which S-shaped patterns are continuously formed.

When the chromium is deposited using the photoresist in which the pattern for forming the chromium coating layer 32 is formed after the gold coating layer 31 is formed, both ends of the chromium coating layer 32 are bonded to the upper portion of the gold coating layer 31 And deposition is carried out so as to connect the gold coating layer 31 to each other.

It is preferable that the gold coating layer 31 has a thickness of 0.1 to 0.3 μm and the chromium coating layer 32 has a thickness of 0.2 to 0.4 μm.

The heater 40 is formed by depositing chromium between the two thermo files 30 in the process of forming the chrome coating layer 32 for forming the thermo file 30. The heater 40 receives AC power to heat the bio sample 2 by AC.

The protective layer 50 prevents the thermopile 30 and the heater 40 from being exposed by covering the thermopile 30 and the heater 40 after the evaporation of the thermopile 30 and the heater 40. The insulating layer 20 and the thermopile 30, And silicon dioxide is deposited on the heater 40.

The thickness of the protective layer 50 from the insulating layer 20 to the top of the protective layer 50 is preferably 0.9 to 1.1 탆 so that the protective layer 50 can sufficiently cover the thermopile 30 and the heater 40 .

The other side of the substrate on which the thermopile 30, the heater 40 and the protective layer 50 are not formed forms a cavity 60 through etching. 1, the heat flow from the heater 40 can be concentrated to the sensor portion by forming the cavity 60 in the lower portion of the substrate portion 10. [

A method of manufacturing the multipurpose micro-calorimeter 1 for a biosample according to the present invention will be described with reference to FIG.

(a) First, the substrate 10 is polished on one or both surfaces of a wafer formed of silicon.

The insulating layer 20 may be formed on both sides of the substrate 10 or on one side of the thermopile 30 and the heater 40 which will be described later.

(b): In order to form the insulating layer 20, silicon dioxide is first deposited on the substrate portion 10 to form the first insulating portion 21. At this time, it is preferable to apply a chemical vapor deposition method (LPCVD) using thermal energy, and the thickness of the first insulation part 21 is formed to be 0.7 to 0.9 mu m.

(c): After forming the first insulating portion 21, the second insulating portion 22 is deposited. The second insulating portion 22 is formed by depositing silicon nitride. The second insulating portion 22 is also preferably formed by impregnating a chemical vapor deposition method using thermal energy. The thickness of the second insulating portion 22 is 0.2 to 0.4 mu m.

The first insulating portion 21 and the second insulating portion 22 are preferably deposited by a chemical vapor deposition method using thermal energy, but a chemical vapor deposition method (PECVD) using plasma energy may also be applied.

After the insulating layer 20 is formed, a thermopile 30 and a heater 40 are formed.

In particular, the thermopile (30) is formed by alternately coating gold and chromium, using a photoresist.

(d): A first photoresist 71 is formed to form a gold coating layer 31 on the insulating layer 20. As shown in the cross-sectional view of FIG. 1, when the gold coating layer is deposited, the gold coating layer is spaced apart from the gold coating layer by a predetermined distance on the first photoresist 71.

(e): The first photoresist 71 is formed so that the gold coating layer 31 is formed, and then gold is deposited to a thickness of 0.3 탆 within 0.1.

(f): After depositing the gold coating layer 31, the photoresist is removed using a cleaning agent. Then, the gold coating layer 31 is deposited on the insulating layer 20 as shown in FIG.

(g): A second photoresist 72 is then formed for the deposition of the chrome coating layer 32. At this time, a pattern for forming the chromium coating layer 32 and a pattern for forming the heater 40 are formed in the second photoresist 72.

(h): chromium is deposited on the second photoresist 72 to a thickness of 0.2 to 0.4 탆, (i) the second photoresist 72 is removed using a cleaning agent, A thermopile 30 comprised of a gold coating layer 31 and a chromium coating layer 32 and a heater 40 positioned between the thermopiles 30 remain.

(j): Silicon dioxide is deposited to a thickness of 0.9 to 1.1 탆 by a chemical vapor deposition method using thermal energy so as to cover the thermopile 30 and the heater 40 on the upper part of the insulating layer 20, And a cavity 60 is formed on the lower side of the substrate unit 10. [ In order to form the cavity 60, a wet etching method using potassium hydroxide (KOH) as an etching solution, for example, a reactive ion etching (RIE) method is used to etch the substrate 10. In addition to potassium hydroxide, hydrochloric acid, nitric acid, sodium hydroxide, sulfuric acid, phosphoric acid, and chromium oxide may be used as the etchant.

The multipurpose micro-calorimeter 1 for a biosample according to the present invention is a micro-calorimeter for biosamples 1, in which a very small amount of biosamples 2 in units of μL are placed on top of a protective layer 50 as shown in FIG. 3, Heat is applied.

Applying an alternating current (ω _v) a heater 40, a heating value oscillates at twice the frequency (2ω _v) by the Joule heating effect.

This can be expressed as follows.

At this time, the thermoelectric voltage signal generated in the thermopile 30 is proportional to the temperature difference between the bio sample 2 and the outer portion of the substrate portion 10, and includes a heat flux signal generated in the bio sample 2.

This can be expressed as follows.

This heat flux signal can be analyzed by separating into an AC component and a DC component at a frequency of 2? _{V. The} AC component represents the change of the heat capacity of the bio sample 2 and the DC component represents the change of the calorific value of the sample. As follows.

For example, when an alternating current of 0.2 mA and 0.1 Hz is applied to the heater 40 as shown in FIG. 4, the heat flux signal sensed through the heater 40 appears as an AC signal of 0.2 Hz offset as shown in FIG.

At this time, if the heat flux signal is frequency-analyzed (FFT), it can be represented by the amplitudes of the DC component and the AC component as shown in FIG. 6, and the change of the calorific value and the heat capacity of the bio sample 2 can be simultaneously measured.

When E. coli is proliferated in culture medium (LB medium) using this method, the increase in the number of individuals and the increase in the calorific value can be simultaneously measured, so that the change in the caloric value due to the increase in the number of individuals can be grasped accurately.

That is, by measuring the concentration of E. coli with time using a UV spectrometer, the growth curve of E. coli as shown in FIG. 7 can be obtained. In this graph, it can be seen that the abrupt increase of population occurs after 7 hours from the start of measurement at room temperature. Based on this result, the same sample was compared with the measurement result through the multi-purpose micro calorimeter (1) for a biosample of the present invention.

As described above, the output signal of the micro calorimeter can be separated into AC signal and DC signal after frequency analysis.

As shown in FIG. 8, the AC signal shows a change in amplitude with time. At the beginning of the measurement, the amplitude of the AC signal increases because the increase in the number of the population is insignificant, but the volume decreases due to evaporation of the culture liquid. However, it can be confirmed that the amplitudes of the AC signals are decreased by the growth of E. coli in the culture medium after the rapid increase of the population.

In addition, the DC signal shows the measurement result as shown in FIG. 9. Since the number of the population is small at the initial stage of measurement, the DC signal decreases due to the latent heat caused by the evaporation of the culture liquid instead of the calorific value of the E. coli. However, As shown in Fig.

The multipurpose micro calorimeter 1 for a biosample according to the present invention can simultaneously measure the number of individuals and the calorific value of the biosample 2 to measure the accurate metabolism of the biosample 2, , Survival rate, growth state, and so on. In the case of liquid biosamples, the latent heat of evaporation and the amount of evaporation can be measured at the same time, so that the influence of evaporation on the calorific value and the heat capacity of the sample can be accurately evaluated.

The description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features presented herein.

1: Multi-purpose micro calorimeter for bio sample
2: Biosample
10:
20: Insulation layer
21: first insulation part 22: second insulation part
30: Thermopile
31: gold coating layer 32: chromium coating layer
40: heater
50: Protective layer
60: cavity
71: first photoresist 72: second photoresist

Claims (15)

A substrate portion polished on one or both surfaces of the wafer;
An insulating layer deposited on the substrate portion and made of two or more different compound layers;
Two thermopiles deposited on the insulating layer to be spaced apart from each other by a predetermined distance;
A heater formed on the insulating layer so as to be positioned between the thermo files and to which an alternating current is applied;
And a protective layer formed on the insulating layer to cover the insulating layer, the thermopile, and the heater,
Wherein the heater is formed to alternately heat the bio sample placed on the top of the protective layer,
The alternating current applied to the heater is used for generating a heat flux signal by the bio sample,
The heat flux signal is divided into an alternating current (AC) signal and a direct current (DC)
Wherein the amplitude change amount of the AC signal and the amplitude change amount of the DC signal are used for simultaneously measuring the number of the biosamples, the heat capacity and the calorific value
Multi-purpose microcalorimeter for bio-samples.
The method according to claim 1,
Wherein the insulating layer includes a first insulating portion formed by depositing silicon dioxide on an upper surface of the substrate portion,
And a second insulating portion formed by depositing silicon nitride on the first insulating portion.
Multi-purpose microcalorimeter for bio-samples.
3. The method of claim 2,
Wherein the first insulating portion has a thickness of 0.7 to 0.9 탆 and the second insulating portion has a thickness of 0.2 to 0.4 탆.
Multi-purpose microcalorimeter for bio-samples.
The method according to claim 1,
Wherein the thermopile comprises a gold coating layer and a chromium coating layer,
Wherein the gold coating layer is spaced apart by a predetermined distance and both ends of the chromium coating layer are laminated on the gold coating layer and deposited on the upper surface of the insulating layer between the gold coating layers
Multi-purpose microcalorimeter for bio-samples.
5. The method of claim 4,
Wherein the gold coating layer has a thickness of 0.1 to 0.3 탆 and the chromium coating layer has a thickness of 0.2 to 0.4 탆.
Multi-purpose microcalorimeter for bio-samples.
The method according to claim 1,
Wherein the protective layer is formed by depositing silicon dioxide, and the thickness from the insulating layer to the top of the protective layer is 0.9 to 1.1 탆.
Multi-purpose microcalorimeter for bio-samples.
The method according to claim 1,
Wherein the substrate portion includes a cavity formed by etching the insulating layer, the thermopile, and the opposed surface opposed to the one surface on which the heater is formed, so as to concentrate heat flow from the heater on the thermopile
Multi-purpose microcalorimeter for bio-samples.
Forming a polished substrate portion on one or both sides of a wafer formed of silicon;
Depositing an insulating layer on at least one side of the substrate portion;
Forming a heater between the two thermopiles spaced apart from each other on the insulating layer and between the thermopiles separated on the insulating layer and applying alternating current power to heat the bio sample with alternating current;
Forming a protective layer on the insulating layer to cover the thermopile and the heater,
The AC power source applied to the heater is used for generating a heat flux signal by the bio sample,
The heat flux signal is divided into an alternating current (AC) signal and a direct current (DC) signal,
Wherein the amplitude change amount of the AC signal and the amplitude change amount of the DC signal are used for simultaneously measuring the number of the biosamples, the heat capacity and the calorific value
A method for manufacturing a multipurpose micro calorimeter for a bio sample.
9. The method of claim 8,
Further comprising the step of forming a cavity by etching the substrate portion on the other side of the substrate portion opposite to the one surface of the substrate portion on which the thermo file and the heater are formed
A method for manufacturing a multipurpose micro calorimeter for a bio sample.
9. The method of claim 8,
The step of depositing the insulating layer may include depositing silicon dioxide on the substrate portion to form a first insulating portion,
And depositing silicon nitride on the first insulating portion to form a second insulating portion
A method for manufacturing a multipurpose micro calorimeter for a bio sample.
11. The method of claim 10,
Wherein the first insulating portion and the second insulating portion are formed by depositing silicon dioxide and silicon nitride using a chemical vapor deposition method using thermal energy, respectively
A method for manufacturing a multipurpose micro calorimeter for a bio sample.
12. The method of claim 11,
Wherein the first insulating portion has a thickness of 0.7 to 0.9 탆 and the second insulating portion has a thickness of 0.2 to 0.4 탆.
A method for manufacturing a multipurpose micro calorimeter for a bio sample.
9. The method of claim 8,
Wherein the step of forming the thermopile comprises forming a gold coating layer on the insulating layer so as to be spaced apart by a predetermined distance and forming a chromium coating layer on both ends of the gold coating layer so as to be laminated on the gold coating layer
A method for manufacturing a multipurpose micro calorimeter for a bio sample.
14. The method of claim 13,
The step of forming the thermopiles may include the steps of forming a first photoresist layer of S (S) pattern on the insulating layer in one direction,
Depositing gold on the first photoresist layer to form a gold coating layer;
Removing the first photoresist layer using a cleaning agent to leave only the gold coating layer on the insulating layer;
Continuously forming a second photoresist layer on the insulating layer, the second photoresist layer connecting the gold coating layers spaced apart from each other;
Depositing chromium on the second photoresist layer to form a chromium coating layer;
And removing the second photoresist layer using a cleaning agent.
A method for manufacturing a multipurpose micro calorimeter for a bio sample.
15. The method of claim 14,
Wherein the heater is formed by depositing chromium on the second photoresist layer so as to be positioned between the thermoresponses when the chromium coating layer is formed by depositing chromium on the second photoresist layer
A method for manufacturing a multipurpose micro calorimeter for a bio sample.

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