Thermal isolation of reaction sites on substrate

Abstract

The invention provides a thermal block assembly. The assembly may comprise: a substrate comprising a first surface configured with a plurality of reaction sites, each reaction site configured for containing a biological sample; and a sample block comprising a plurality of susceptors configured to thermally condition the plurality of biological samples, where each susceptor is thermally coupled to one of the reaction sites. The assembly may also include a cooling block, a slot, and an insulating ring associated with the reaction sites, each of the cooling block, the slot, and the insulating ring being capable of minimizing heat flow between the reaction sites.

Description

[0001] This application is a divisional application of the invention patent application with application number 20168056544.7, application date September 2, 2016, and invention name “Thermal isolation of reaction sites on substrate”. Background Art

[0002] Polymerase chain reaction (PCR) is a common tool for amplifying DNA samples in genetic analysis. The process may include placing a DNA biological sample in a sample holder or on a sample holder, which is suitable for isolating each sample from other samples at the reaction site. Such sample holders are well known in the art and can take various forms, such as but not limited to microtiter plates, microcards, separate tubes, connecting tube strips, capillaries, microarrays and slides. In addition, the number of samples accommodated in the sample holder can vary according to, for example, the required analysis type, and a single sample holder or a sample holder can accommodate one sample to several dry samples.

[0003] One of the challenges encountered in designing sample holders for multiple functions such as mixing, fluid transfer, heating and cooling is that it is difficult to thermally control a certain area of ​​the holder without thermally affecting adjacent areas. It can also be challenging in the case of processing small, closely spaced samples, such as polymerase chain reaction (PCR). Amplifying DNA (deoxyribonucleic acid) using the PCR process involves cycling a specially composed liquid reaction mixture through several different incubation periods in a thermal cycler. The reaction mixture consists of various components, including the DNA to be amplified and at least two primers that are sufficiently complementary to the sample DNA so that extension products of the amplified DNA can be produced. The key to PCR is the concept of thermal cycling in a thermal cycling instrument. The thermal cycler can be designed to perform alternative steps to denature the DNA, anneal short primers to the resulting single strands, and extend those primers to form new copies of the double-stranded DNA. During the thermal cycling process, the PCR reaction mixture is repeatedly cycled between a high temperature of about 95°C for denaturing the DNA to a low temperature of about 50°C to 70°C for primer annealing and extension.

[0004] In some previous PCR instruments, the sample tube is inserted into the sample hole on the metal block. To implement the PCR process, the temperature of the metal block is cycled according to the temperature and time specified by the user in the PCR protocol file. The cycle is controlled by a computer and associated electronic devices. As the temperature of the metal block changes, the temperature of the samples in each tube changes similarly. Such heated metal blocks can be used with any of the above-mentioned sample holders. However, when, for example, PCR is performed on very small samples, the device may be subject to very harsh thermal cycle schemes, which are not common in some previous PCR instruments. One such scheme may require one or more samples to be heated to, for example, 95°C, while one or more adjacent samples need to be maintained at a temperature that is greatly different from 95°C. With respect to samples that require different temperatures, the block can not only heat the desired reaction site, but also transfer heat to the surrounding area and other samples. This heat transfer can have an adverse effect on other samples and have a negative impact on the amplification or incubation of the sample. Therefore, it is desirable to provide a sample holder and can be used to perform, for example, PCR thermal cycle reactions without thermal effects on adjacent reactions. Summary of the invention

[0005] In one embodiment of the present invention, a thermal block assembly is provided. The assembly includes: a substrate including a first surface configured with a plurality of reaction sites, each reaction site configured to accommodate a biological sample, wherein the substrate is configured with a feature structure to improve thermal isolation of the reaction site; and a sample block including a plurality of pedestals configured to thermally condition a plurality of biological samples, wherein each pedestal is thermally coupled to one of the reaction sites.

[0006] In another embodiment, a thermal block assembly is provided. The assembly includes: a base including a first surface configured with a plurality of reaction sites, each reaction site configured to receive a biological sample; a sample block including a plurality of pedestals configured in one or more rows, wherein each pedestal is thermally coupled to one of the reaction sites and configured to have a feature structure to improve thermal isolation of the reaction sites; and a plurality of cooling blocks, each cooling block being associated with a reaction site and capable of minimizing heat flow between the reaction sites.

[0007] In another embodiment, a method for thermally isolating reaction sites is provided. The method includes the following steps: providing a substrate including a plurality of reaction sites, each reaction site being configured to receive a biological sample; providing a sample block including a pedestal, each pedestal having a size substantially equal to the size of the reaction site and being thermally coupled to the reaction site; using a thermoelectric device to adjust the temperature of the pedestal through a series of temperatures and hold times; thermally isolating the reaction sites from each other; and cooling the reaction sites using a cooling block.

[0008] The following description and claims set forth additional aspects, features and advantages of the present invention, particularly when considered in conjunction with the accompanying drawings in which like reference numerals represent like components.

[0009] BRIEF DESCRIPTION OF THE DRAWINGS

[0010] For a more complete understanding of the principles disclosed herein and its advantages, reference is now made to the following description taken in conjunction with the accompanying drawings, in which:

[0011] Figure 1 is a perspective view of a substrate with two rows of reaction sites and their spatial relationship to a sample block.

[0012] Figure 2A A thermal analysis perspective of a row of reaction sites heated by a single heating block or rod.

[0013] Figure 2B Thermal perspective of a row of heated reaction sites and their thermal effects on adjacent rows.

[0014] Figure 3A is a perspective view of a row of reaction sites heated by a pedestal including a thermal block according to various embodiments.

[0015] Figure 3B A perspective view of the effect of one row of heated reaction sites on an adjacent row.

[0016] Figure 4 is a perspective view of a heating block including pedestals and a radius added to each pedestal according to various embodiments.

[0017] Figure 5 To pass Figure 4 Thermal map of a row of reaction sites heated by a heating block.

[0018] Fig. 6A is a top view of a substrate having a slot inserted between two rows of reaction sites according to various embodiments.

[0019] Figure 6B A bottom view illustrating the relationship between a sample block and a slot inserted between two rows of reaction sites according to various embodiments.

[0020] Fig. 7A is a perspective view of a substrate having a slot inserted between two rows of reaction sites and offset with respect to the reaction sites according to various embodiments.

[0021] Figure 7B A perspective thermal analysis of rows of heated reaction sites and adjacent rows of reaction sites with slots interposed between the rows.

[0022] Figure 8is a perspective view of a substrate having increased spacing between rows according to various embodiments.

[0023] Fig. 9A A thermal analysis perspective view of a heated reaction site row and adjacent reaction site rows.

[0024] Fig. 9B Another perspective thermogram of a heated row of reaction sites and adjacent rows of reaction sites.

[0025] Fig. 10A Another perspective thermal analysis of a row of reaction sites and adjacent rows of reaction sites heated according to various embodiments.

[0026] Fig. 10B Another perspective thermogram of a heated row of reaction sites and adjacent rows of reaction sites.

[0027] Fig.11 is a perspective view of a thermal block according to various embodiments, with a cooling block added below an adjacent reaction site.

[0028] Fig.12 A thermal analysis perspective view of a heated reaction site row and adjacent reaction site rows. DETAILED DESCRIPTION

[0029] Embodiments of apparatus, systems, and methods for providing thermal isolation between elements of a substrate are described herein.The section headings used herein are for organizational purposes only and are not to be construed as limiting the described subject matter in any way.

[0030] Reference will now be made in detail to various aspects of the present invention, examples of which are illustrated in the accompanying drawings. Wherever possible, the same reference numerals will be used throughout the drawings to refer to the same or like parts.

[0031] In this detailed description of various embodiments, for purposes of explanation, numerous specific details are set forth to provide a thorough understanding of the disclosed embodiments. However, those skilled in the art will appreciate that these various embodiments may be practiced with or without these specific details. In other cases, structures and devices are presented in block diagram form. Furthermore, those skilled in the art can readily appreciate that the specific order in which the methods are presented and performed is illustrative, and it is contemplated that the order may be changed and still remain within the spirit and scope of the various embodiments disclosed herein.

[0032] All literature and similar materials cited in this patent application (including but not limited to patents, patent applications, articles, books, papers, and Internet web pages) are expressly incorporated herein by reference in their entirety for any purpose. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which the various embodiments described herein belong. When the definition of a term in an incorporated reference appears to differ from the definition provided in the present teachings, the definition provided in the present teachings shall prevail.

[0033] It should be understood that "about" is implied before the temperatures, distances, etc. discussed in the present teachings, so that minor and insubstantial deviations are within the scope of the present teachings. In this patent application, unless otherwise specifically stated, the use of the singular includes the plural. In addition, the use of "including", "containing" and "comprising" is not intended to be limiting. In this patent application, the term "substrate" is used to refer to all sample holder forms known in the art and is not intended to be limited to any particular form. In addition, it should be understood that the above general description and the following detailed description are merely exemplary and explanatory and do not limit the present teachings.

[0034] Although the present teaching content is described in conjunction with various embodiments, it is not intended to limit the present teaching content to such embodiments. On the contrary, it should be understood by those skilled in the art that the present teaching content encompasses various alternatives, modifications and equivalent forms.

[0035] In addition, when describing various embodiments, this specification may have presented the method and / or process as a specific sequence of steps. However, to the extent that the method or process does not rely on the specific order of steps set forth herein, the method or process should not be limited to the specific sequence of steps described. It should be understood by those of ordinary skill in the art that other sequences of steps may be possible. Therefore, the specific order of steps set forth in this specification should not be interpreted as a limitation on the claims. In addition, the claims for the method and / or process should not be limited to the execution of its steps in the order written, and those skilled in the art can easily understand that the sequence can be varied and still remain within the spirit and scope of the various embodiments.

[0036] The present teachings disclose various embodiments of substrates and sample blocks with improved thermal isolation throughout the device. As described in detail below, various embodiments of substrates and sample blocks with improved thermal isolation provide desirable performance for bioanalytical instruments utilizing such devices.

[0037] For example, in the case of PCR, it is desirable to change the sample temperature as quickly as possible between the temperatures required in the cycle for several reasons. First, each stage of the chemical reaction has its optimal temperature, so spending less time at non-optimal temperatures can mean obtaining better chemical results. Second, a minimum time is usually required at any given set point that sets the minimum cycle time for each method, and any time spent in transition between set points is added to this minimum time. Since the number of cycles is usually quite large, this transition time can significantly increase the total time required to complete the amplification.

[0038] The absolute temperature of each reaction site obtained in each step of the method is critical to the yield of the product. Therefore, it is advantageous to thermally isolate regions of the substrate in order to minimize the effect of one region on another. The geometries that can be seen in the substrate are often small, resulting in hot or warm regions being very close to cooler regions of the device. Since heat flows through the device from the hot or warm regions to the cool regions, this close proximity can result in higher temperatures in the cooler regions than expected, thereby affecting the performance of the cooler regions. In addition, small geometries make thermal isolation of hot and cool regions difficult to achieve.

[0039] Figure 1 An embodiment not uncommon in the art is shown. In this embodiment, a substrate 140 is shown, with two rows 110 and 120 eight reaction sites on the substrate. A heating block 130 is connected to the lower side of the substrate 140, and the heating block is in the shape of a rod, located below the row 110 and opposite to the substrate of the row 100. Such a relationship between the substrate, the reaction site and the heating block is also not uncommon in the art. The heating block is also well known in the art and is usually made of a material with high thermal conductivity. Some examples of suitable materials for the heating block include, but are not limited to, silver, gold, aluminum, magnesium and copper. In some embodiments, the heating block can be made into a rectangular geometric shape, but it can also be round, square or any other shape suitable for related applications. Typically, the surface of the block has an array of depressions that can receive sample container devices sometimes referred to as plates or tubes. The array size varies, and can be, for example, 8, 16, 24, 48, 96, 384, 1536 or more. In some applications, the metal block may include a flat surface that lacks recesses for accommodating an array of reaction sites that may have a flat surface such as a glass slide.

[0040] In this embodiment, block 130 is fabricated as a metal bar with a flat upper surface and is sized to be thermally coupled to the underside of substrate 140 opposite the reaction sites in row 110. Row 120 is maintained at room temperature or approximately 27° C. Block 130 is heated to 95° C. and determines the temperature of rows 110 and 120. Figure 2AThe results of the thermal analysis are shown. It is apparent from this thermal map that rows 110 and 120 are both heated to almost the same temperature and the individual reaction sites are indistinguishable. Figure 2B A more detailed interaction between heated row 110 and unheated row 120 is shown. As shown by the uniform shading, heated row 110 has a uniformly high temperature. This temperature provides a high enough temperature difference between rows 110 and 120 that heat flows from row 110 through substrate 140 and to row 120. Figure 2B The temperature scale included in indicates that the temperature of the area of ​​the unheated row 120 closest to the heated row 110 is about 50°C. In addition, it can also be seen from the figure that there is a significant temperature gradient within the unheated row 120 of about 20°C. These results demonstrate the magnitude of the interaction of closely spaced adjacent rows with significantly different temperature requirements. When performing PCR, it is common (for example) to heat a row to 95°C and hold it for up to 10 minutes, while requiring the temperature of the adjacent row to remain below 37°C. To ensure that the heated row does not produce adverse thermal interactions with the unheated adjacent row, it would be beneficial to minimize the thermal effects on the adjacent row. Figure 2A and Figure 2B The data presented confirms thermal interaction between adjacent rows and suggests a need for embodiments that minimize this effect.

[0041] Many factors can affect the thermal interaction between the reaction sites. Some of these factors may include, but are not limited to, spacing, reaction site size that may be related to spacing, and temperature differences between the reaction sites and the substrate material. It will be appreciated by those skilled in the art that one or a combination of these factors may be required to achieve the desired degree of thermal insulation. With this understanding, a solution for thermal isolation is provided below.

[0042] Base material

[0043] The material used to make the substrate can be, for example, an elastomer, such as a polymer that exhibits rubber-like elasticity. Those skilled in the art will also appreciate that elastomers include many types, such as saturated rubbers, unsaturated rubbers, thermoplastic elastomers, and polysulfide rubbers. These materials all exhibit relatively poor thermal conductivity and therefore provide a high level of thermal insulation between adjacent rows of reaction sites. For example, Thermally conductive elastomers exhibit thermal conductivities as high as 5 W / mK. In comparison, metals such as aluminum, copper, gold, and silver exhibit thermal conductivities of 205 W / mK to 406 W / mK. However, as described above, even though they are poor thermal conductors, the heat conducted is still sufficient to affect the temperature of the reaction in the adjacent pores of the substrate.

[0044] The substrate may also be made of a polymer including, for example, polypropylene, although other polymers may also be used. By way of example, polypropylene has a thermal conductivity of approximately 0.1 W / mK to 0.22 W / mK. This thermal conductivity may still result in a reaction site affecting other reaction sites if the temperature difference between the reaction sites is large enough (as described above).

[0045] Heat block geometry

[0046] from Figure 2B As can be clearly seen in the thermal map of FIG. 1 , heating the sample block 130 to 95° C. provides sufficient heat to the row 110 and substrate 140 to heat at least a portion of the row 120 to approximately 50° C., which, as described above, would result in sufficient heat to adversely affect the reaction in the row 120. Figure 3A Shown with Figure 2A The heatmap shown is similar to the heatmap shown in Figure 3A In the embodiment shown, Figure 2B The block 130 in the figure is replaced by a block 150. The difference between the block 150 and the block 130 is that the flat upper surface of the block 130 has been replaced by a series of bases. The upper surface of each base may have a substantially circular shape (such as Figure 3A ), square, oval, rectangular or any other shape suitable for the relevant application. In addition, the upper surface 450 of each base (as shown) Figure 4 4 (shown in FIG. 4 ), for example, may have an area substantially equal to, less than, or greater than the area of ​​the contact surface of the corresponding reaction site. Regardless of the shape and size, the block 150 may be located below the substrate 140, wherein each base surface 450 (such as Figure 4 ) and an area of ​​the base surface 620 (as shown Figure 6B ), which is opposite to the reaction site.

[0047] The heat block 150 was set to 95°C and the results were as follows Figure 3A shown. Figure 3A Clearly shown in Figure 2A The addition of a susceptor to heat block 150 demonstrates that more localized heating of the reaction sites in row 110 is achieved without significantly affecting row 120. Figure 3B More detailed thermal analysis results are provided for rows 110 and 120. Again, row 110 is shown as having a uniform shading, indicating that its temperature is uniform. Figure 3B The temperature scale provided in now indicates that the hottest region of row 120 is now about 46°C, and the gradient between reaction sites is about 16°C. Comparing these results with Figure 2AThe results shown are compared to those shown above, which show that adding the susceptor to the thermal block 150 reduces the absolute temperature of the row 120 and improves the thermal gradient in the reaction site by about 4°C or about 8%. Those skilled in the art will recognize that the size and geometry of the susceptor and the reaction site dimensions may be modified for other applications, as described above.

[0048] Compared to Figure 2A Another embodiment of improving thermal isolation between adjacent reaction sites is Figure 4 . The heat block 440 shown has a base 430 and a base surface 450. In this example, the base is substantially circular in shape. However, those skilled in the art will recognize that the base can be of any shape or size, as described above. In this case, the base has been modified to include a radius around the top circumference to accommodate rings 410 and 420. The ring 410 shown has a thickness that is less than the ring 420 and is substantially circular to match the shape of the base. Those skilled in the art will appreciate that the rings can be of any shape or size that matches the shape of the base, and therefore may not be described as rings in some embodiments. Rings 410 and 420 may also include insulating material to further prevent heat from leaking from the heat block 440 to adjacent rows. The results obtained by heating the heat block 440 to 95°C and maintaining it for 10 minutes are shown as follows. Figure 5 As shown. Figure 5 On the temperature scale shown, row 120 is heated by row 110 to a maximum temperature of about 43°C. Figure 5 It is also shown that the temperature of the pedestal with ring 410 is higher than the pedestal with ring 420, thereby confirming that changing the geometry of the pedestal and including the thermal resistance ring reduces the thermal interaction between the rows. In this example, the unheated row 120 reaches a maximum temperature of about 43°C with a thermal gradient of 14°C, thereby comparing to the Figure 2A The data in gives an overall improvement of 14% in the temperature of row 120.

[0049] The slot between the reaction sites

[0050] Fig. 6A Another embodiment is shown where a slot 610 is located between rows 110 and 120. The slot 610 can introduce thermal resistance between the rows, thereby reducing the amount of heat flowing from the heated rows to the unheated rows. Factors that can affect the flow of heat in a device can include, but are not limited to, the length of the gap, the width of the gap, and the location of the gap. Fig. 6A A perspective view of a portion of a substrate is shown, which includes a heated block 150 located below the substrate 140 and aligned opposite the reaction sites in the row 110. The block 150 is aligned with the Figure 4 The block 150 has the same construction. Fig. 6AAlso shown are the reaction sites in each row 110 and the slots between rows 120. The distance between rows 110 and 120 is 10 mm, and the slots are 1 mm x 4 mm in size. Those skilled in the art will also appreciate that the size of the slots and the distance between rows are arbitrary and are selected to provide the desired performance of this embodiment. In this example, the distance between rows and slots is based on the size of the substrate and the corresponding reaction sites, as well as the density of the reaction sites.

[0051] To determine the effect of this embodiment, block 150 was heated to 95°C and held at that temperature for 10 minutes. As mentioned above, this time / temperature combination is not uncommon in PCR protocols and may represent a worst-case scenario for heat leakage into adjacent reaction sites. Figure 6B The results of the thermal analysis are shown along with a view of the underside of substrate 140. Block 150 is oriented so that the upper four pedestals are fitted with a 0.5 mm thick ring and the lower four pedestals are fitted with a 1.0 mm thick ring. Slots 610 prevent heat from flowing from row 110 to row 120 so that the maximum temperature of the upper 4 reaction sites of row 120 is in the range of 36°C-38°C and the maximum temperature of the lower 4 reaction sites is in the range of 35°C-37°C.

[0052] Figure 6B The results shown show that Figure 2A In contrast, using narrow slots between reaction sites can also be an effective way to block heat flow, thereby significantly reducing the maximum temperature of adjacent rows. It will be apparent to those skilled in the art that the slots can be further optimized. Fig. 7A One such embodiment shown in shows grooves that are slightly offset between reaction sites. Figure 7B The results of thermal analysis obtained by heating the heat block to 95°C and holding it at 95°C for 10 minutes are shown. As shown in the temperature scale, the maximum temperature of the adjacent reaction site has dropped to about 34°C. This result shows the Figure 2A Improves the thermal isolation effect between reaction sites compared to narrow grooves.

[0053] Row spacing and ambient temperature

[0054] According to another embodiment, with respect to Figure 1 , thermal isolation can be improved by varying the distance between rows of reaction sites.

[0055] Figure 8 The intervals shown are relative to the above Figure 1 After the heating block 150 is heated to 95°C and maintained at 95°C for 10 minutes (ambient temperature 25°C), Fig. 9A Results obtained after a 10 minute hold are shown. Fig. 9AThe temperature of the adjacent row 120 reaches a maximum temperature of about 26°C. This maximum temperature is compared to Figure 1 The spacing shown significantly improves thermal isolation. Fig. 9B , the effect of increasing from ambient temperature to 30° C. is shown. Referring to the temperature scale therein, a 5° C. increase in ambient temperature results in a temperature increase of approximately 4° C. in row 120 . Fig. 10A shows similar results for a row spacing of 15 mm at an ambient temperature of 25°C, and Fig. 10B The results obtained with the same 15 mm spacing at an ambient temperature of 30°C are shown. This data shows that the increase in spacing between reaction sites and the decrease in ambient temperature compared to Figure 2A Thermal isolation between reaction sites may be improved.One skilled in the art will recognize that the row spacing selected may depend on the geometry of the substrate as well as the reaction site density.

[0056] Cooling block

[0057] In another embodiment, adjacent rows as described above may be cooled. Fig.11 One embodiment is shown on the underside of substrate 140. Heat blocks 150 are also shown opposite the reaction sites in row 110, as previously described, and cooling blocks 1110 are shown opposite the reaction sites in row 120, with one cooling block aligned with each reaction site.

[0058] In one embodiment, the cooling block 1110 can be used to provide greater thermal mass to adjacent rows 120 in much the same manner as a heat sink can be used to transfer heat from a high temperature object to the environment. Therefore, it would be advantageous for the cooling block to include a thermally conductive material such as aluminum and copper. It will be appreciated by those skilled in the art that the effectiveness of the cooling block in this embodiment may also depend on, for example, the relative size of the cooling block 1110 and the size of the reaction site.

[0059] An alternative embodiment can provide active cooling for the cooling block 1110. Active cooling can be provided by any of a variety of implementations known in the art. For example, various implementations of active cooling can include, but are not limited to, thermoelectric cooling, pumping a coolant through the cooling block, and heat pipes. The thermal analysis results of the active cooling solution are shown in FIG. Fig.12 The heating block 150 was heated to 95°C and maintained at 95°C for 10 minutes, and the cooling block 1110 was cooled and maintained at 25°C, and then the collected Fig.12 As can be seen from the temperature scale, the hottest edge of row 120 remains at approximately 28°C, which is Figure 2A The maximum temperature of the middle line is about half of 120. Fig.12 It can also be seen that compared with Figure 2AAs shown in the thermal map, the thermal gradient in adjacent rows 120 has been significantly reduced, further demonstrating the effectiveness of the cooling blocks 610 in thermally isolating the reaction sites.

[0060] Although the principles of the present invention have been described in conjunction with various embodiments of heat blocks and substrates, it should be clearly understood that these descriptions are for example only and are not intended to limit the scope of the present invention. The content disclosed herein has been provided for the purpose of illustration and description. This is not intended to be exhaustive or to limit the disclosed content to the precise form described. Many modifications and variations will be apparent to those skilled in the art. The disclosed content is selected and described in order to best explain the principles and practical applications of the disclosed embodiments in the field, thereby enabling other technical personnel in the field to understand the various embodiments and various modifications suitable for the specific purposes covered. The scope of the present disclosure is intended to be limited by the following claims and their equivalents.

Claims

1. A heat block assembly comprising: a substrate comprising a substrate surface configured with a plurality of reaction sites, each reaction site being configured to receive a biological sample; Sample block; as well as Thermally insulating features configured to limit heat flow from one reaction site to another reaction site.

2. The block assembly according to claim 1, characterized in that The thermally isolating features include features selected from the group consisting of a base, a slot, a cooling block, a ring, a reaction site spacer, and combinations thereof.

3. Block assembly according to any one of the preceding claims, characterized in that The thermal isolation feature includes a base and is disposed on the sample block.

4. Block assembly according to any one of the preceding claims, characterized in that The insulating features include a base, and the top of each base includes a base surface.

5. Block assembly according to any one of the preceding claims, characterized in that The insulating features include a base, and a top of each base includes a base surface, wherein each base surface is adjacent to the base surface.

6. Block assembly according to any one of the preceding claims, characterized in that The thermal isolation feature comprises a base, wherein the base and the reaction site have substantially the same shape, wherein the shape is selected from the group consisting of circular, oval, rectangular, and square.

7. Block assembly according to any one of the preceding claims, characterized in that The thermal isolation feature comprises a pedestal, wherein each pedestal and reaction site have substantially the same shape, and wherein each pedestal and reaction site are substantially circular.

8. Block assembly according to any one of the preceding claims, characterized in that The thermal isolation features include a base, and each base has a size substantially equal to a size of each reaction site.

9. Block assembly according to any one of the preceding claims, characterized in that The insulating features include a base, and the top of each base includes a base surface, wherein each base includes an insulating feature including a ring mounted around the base surface at a radius.

10. Block assembly according to any one of the preceding claims, characterized in that The insulating features include a base, and the top of each base includes a base surface, wherein each base includes an insulating feature, the insulating feature including a ring mounted around the base surface at a certain radius, wherein each ring is composed of an insulating material.

11. Block assembly according to any one of the preceding claims, characterized in that The thermal isolation features include cooling blocks, each cooling block being positioned adjacent one of the reaction sites.

12. Block assembly according to any one of the preceding claims, characterized in that The thermally insulating feature comprises a cooling block, and wherein the cooling block is comprised of a thermally conductive material.

13. Block assembly according to any one of the preceding claims, characterized in that The thermal isolation feature comprises a cooling block, and wherein the cooling block is passively cooled.

14. Block assembly according to any one of the preceding claims, characterized in that The thermal isolation feature comprises a cooling block, and wherein the cooling block is actively cooled.

15. Block assembly according to any one of the preceding claims, characterized in that The thermal isolation feature includes a slot, wherein the slot is disposed in a substrate between reaction sites and extends through the substrate.

16. Block assembly according to any one of the preceding claims, characterised in that The thermally insulating feature comprises a slot, and the slot is rectangular.

17. A block assembly according to any one of the preceding claims, characterised in that The thermally insulating feature comprises a slot, and the slot is curved.

18. A block assembly according to any one of the preceding claims, characterised in that The thermally insulating feature structure includes a reaction site spacing.

19. A block assembly according to any one of the preceding claims, characterised in that The sample block is composed of a thermally conductive material.

20. A method for thermally isolating a reaction site, the method comprising: providing a substrate comprising a plurality of reaction sites, each reaction site being configured to receive a biological sample; providing a sample block comprising a plurality of pedestals, each pedestal having a size substantially equal to a size of the reaction site and thermally coupled to the reaction site; thermally isolating the reaction sites from each other using thermally insulating features; regulating the temperature of the susceptor through a series of temperatures and hold times; and cooling the reaction site using a cooling block.

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