Biochip, biological detection system and biological detection method

Abstract

The embodiment of the present invention provides a biochip, a biodetection system and a biodetection method, wherein the biochip includes a first substrate, a polarizing array and a plurality of reaction zones. The polarizing array is disposed on the first substrate, and the polarizing array includes a first polarizing unit and a second polarizing unit, the first polarizing unit has a first polarization angle, and the second polarizing unit has a second polarization angle, and the first polarization angle is different from the second polarization angle. The reaction zones are disposed on the polarizing array. Each reaction zone corresponds to one of the first polarizing unit or the second polarizing unit.

Description

Technical Field

[0001] Embodiments of the present invention relate to a biochip, a biological detection system, and a biological detection method, and particularly to a biochip having a polarization array and a biological detection system having a front polarizing element. Background Art

[0002] Integrated sensing devices have recently been used in bioanalysis. When using such applications, biological or biochemical samples are placed on a biochip. The phenomenon of a biological reaction or interaction can be reported by the emission spectrum and / or intensity of fluorescent molecules, such as DNA sequencing and immunofluorescence detection. The fluorescent molecules can be excited by excitation light of a short wavelength and generate emission light of a longer wavelength toward a light detector. The spectral distribution and intensity of the fluorescence can be detected and judged by the light detector of the biological detection system.

[0003] In the process of the evolution of biochips, in order to pursue lower costs and achieve higher production capacity, the array density on the biochip is generally increased by reducing the spacing width or well pitch. However, reducing the array size may cause crosstalk between adjacent wells and may not be able to accurately detect each individual fluorescent signal, thereby resulting in inaccurate analysis results.

[0004] Although existing biochips generally meet their requirements, they are not satisfactory in all aspects. Therefore, there is still a need for a new biochip, a new biological detection system, and a biological detection method. Summary of the Invention

[0005] According to some embodiments of the present invention, a biochip is provided. The biochip includes a first substrate, a polarization array, and a plurality of reaction regions. The polarization array is disposed on the first substrate. The polarization array includes a first polarization unit and a second polarization unit. The first polarization unit has a first polarization angle, and the second polarization unit has a second polarization angle, wherein the first polarization angle is different from the second polarization angle. The reaction regions are disposed on the polarization array. Each of the reaction regions corresponds to one of the first polarization unit or the second polarization unit.

[0006] According to some embodiments of the present invention, a biological detection system is provided. The biological detection system includes an excitation light source, a front polarizing element, the above-mentioned biochip, a lens, and a light detector. The excitation light source is used to emit excitation light. The front polarizing element is used to polarize the excitation light. The biochip is used to receive the polarized excitation light. The lens is used to concentrate the emission light, and the emission light is from a biological sample fixed in the reaction region of the biochip. The light detector is used to detect the emission light.

[0007] According to some embodiments of the present invention, a biological detection method is provided. The biological detection method includes the following steps: providing the above-mentioned biological detection system; fixing a biological sample to a reaction area of a biochip; performing a first detection step to obtain a first fluorescence signal, the first fluorescence signal being from the biological sample fixed to the reaction area corresponding to the first polarization unit; performing a second detection step to obtain a second fluorescence signal, the second fluorescence signal being from the biological sample fixed to the reaction area corresponding to the second polarization unit; and combining the first fluorescence signal and the second fluorescence signal.

[0008] In the following embodiments, a detailed description is provided with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Reading the following detailed description and examples in conjunction with the accompanying drawings will enable a more comprehensive understanding of the embodiments of the present invention, wherein:

[0010] Figures 1A to 1C is a cross-sectional view of a biological detection system according to some embodiments of the present invention.

[0011] Figure 1D is a cross-sectional view of a biochip according to some embodiments of the present invention.

[0012] Figure 1E is a cross-sectional view of a biological detection system according to other embodiments of the present invention.

[0013] Figures 2A to 2B is a top view of a biochip according to some embodiments of the present invention.

[0014] Figure 3 is a flowchart of a biological detection method according to some embodiments of the present invention.

[0015] Figures 4A to 4B illustrates the use of a biological detection system to detect a biological sample according to some embodiments of the present invention.

[0016] Among them, the reference numerals are explained as follows:

[0017] 10A, 10B, 10C: Biological detection system

[0018] 100A, 200: Biochip

[0019] 102: First substrate

[0020] 104: Polarization array

[0021] 104A: First polarization unit

[0022] 104B: Second polarization unit

[0023] 106: Sample isolation layer

[0024] 108: Reaction region

[0025] 110: Front polarizing element

[0026] 111: Excitation light source

[0027] 112: Excitation light

[0028] 112’: Polarized excitation light

[0029] 114: Lens

[0030] 116: Filter element

[0031] 118: Light detector

[0032] 120: Planarization layer

[0033] 122: Second substrate

[0034] 124: Spacer layer

[0035] 126: Microfluidic channel

[0036] 128: Opening

[0037] 130: Spectral element

[0038] 132: Emitted light

[0039] 300: Biological detection method

[0040] 302, 304, 306, 308, 310: Steps

[0041] 400A, 400B: Biological samples

[0042] 400AS: First fluorescence signal

[0043] 400BS: Second fluorescence signal

[0044] A - A’: Section line Detailed implementation manners

[0045] The following details the biochip, biological detection system and biological detection method of the embodiments of the present invention. For the purpose of illustration, many specific details and embodiments are described in the following detailed description to fully understand the embodiments of the present invention. The specific elements and configurations described in the following detailed description are used to clearly describe the embodiments of the present invention. However, the exemplary embodiments described herein are clearly only for illustration, and the concepts of the embodiments of the present invention can be presented in various forms and are not limited to these exemplary embodiments.

[0046] In addition, for a clear description of the embodiments of the present invention, similar and / or corresponding numbers may be used in the illustrations of different embodiments to represent similar and / or corresponding elements. However, this does not imply any relationship between different embodiments. It should be understood that the description of this exemplary embodiment can be understood in conjunction with the illustrations, and the illustrations of the embodiments of the present invention are also regarded as part of the description of the embodiments of the present invention. The drawings are not drawn to the scale of actual devices and elements. In addition, the structures and devices are illustrated in a schematic manner to simplify the drawings.

[0047] In addition, when referring to "a film layer is overlying another film layer", "a film layer is above another film layer", "a film layer is on another film layer", or "a film layer is over another film layer", it may refer to the case where the film layer is in direct contact with other film layers. Or, it may also be the case where the film layer is not in direct contact with other film layers, in which case one or more intermediate layers are provided between the film layer and other film layers.

[0048] In addition, relative terms are used in this specification. For example, "lower", "bottom", "higher", or "top" are used to describe the relative position of one element with respect to another element. It should be understood that if the device is flipped upside down, the element described on the "lower" side will become the element on the "higher" side.

[0049] It should be understood that although terms such as "first", "second", "third", etc. may be used herein to describe various elements, components, regions, film layers, and / or parts, these elements, components, regions, film layers, and / or parts should not be limited by these terms. These terms are only used to distinguish different elements, components, regions, film layers, and / or parts. Therefore, the first element, component, region, film layer, and / or part discussed below may be referred to as the second element, component, region, film layer, and / or part without departing from the teachings of the embodiments of the present invention.

[0050] The terms "about" and "substantially" generally mean within 10% of a given value or range, preferably within 5%, or within 3%, or within 2%, or within 1%, and more preferably within 0.5%. The quantity given herein is an approximate quantity, that is, the meaning of "about" or "substantially" may still be implied even without specific mention of "about" or "substantially".

[0051] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It should be understood that these terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning consistent with the relevant art and the background or context of this invention, and should not be interpreted in an idealized or overly formal manner, unless specifically defined in the embodiments of this invention.

[0052] According to some embodiments of the present invention, the polarization array of the biochip may include a first polarization unit and a second polarization unit. The first polarization unit has a first polarization angle and the second polarization unit has a second polarization angle. The first polarization angle is different from the second polarization angle. In addition, according to some embodiments of the present invention, the biological detection system may include a front polarization element, which is orientable by rotation so that the front polarization element has the same polarization angle as the first polarization angle or the second polarization angle. With such a configuration in combination with a two-step detection method, crosstalk between fluorescence signals in adjacent reaction regions can be reduced, and the biochip can be fabricated with a higher array density.

[0053] Figures 1A to 1B is a cross-sectional view of the biological detection system 10A according to some embodiments of the present invention. It should be understood that according to some embodiments of the present invention, additional components may be added to the biological detection system 10A.

[0054] Refer to Figure 1A, the biological detection system 10A includes a biochip 100A. The biochip 100A includes a first substrate 102, a polarization array 104, and a plurality of reaction regions 108 defined between sample isolation layers 106. The first substrate 102 can include any suitable material. In some embodiments, the first substrate 102 can be a flexible material, such as polyethylene terephthalate (PET), polyethersulfone (PES), polyimide (PI), polycarbonate (PC), polymethylmethacrylate (PMMA), silicone, epoxy resin, or a combination of the foregoing. In other embodiments, the first substrate 102 can be a rigid material, such as a glass substrate or a sapphire substrate. Furthermore, the first substrate 102 can be transparent or translucent. Specifically, in embodiments where the first substrate 102 is transparent, the material of the first substrate 102 can have a light transmittance greater than 85% for light with a wavelength between 400 nm and 750 nm, or preferably greater than 92% light transmittance. In embodiments where the first substrate 102 is translucent, the material of the first substrate 102 can have a light transmittance greater than 25% and less than 85% for light with a wavelength between 400 nm and 750 nm.

[0055] The polarization array 104 is disposed on the first substrate 102. In some embodiments, the polarization array 104 can include a first polarization unit 104A and a second polarization unit 104B. The first polarization unit 104A has a first polarization angle, and the second polarization unit 104B has a second polarization angle. The term "polarization angle" as used herein refers to light that is polarized at an angle of 90° or polarized at an angle perpendicular to the polarization angle that cannot pass through the sub-polarization unit having this polarization angle. The absolute value difference between the first polarization angle and the second polarization angle can be greater than 0° and less than 180°, such as 0°, 45°, 90°, 135°, or 180°. In some specific embodiments, the absolute value difference between the first polarization angle and the second polarization angle can be 90°. Although in Figure 1A , the polarization array 104 of the biochip 100A is depicted as having two types of sub-polarization units, but the present invention is not limited thereto. In other embodiments, the polarization array 104 can further include a third polarization unit and even a fourth polarization unit (not shown), and the third polarization unit and the fourth polarization unit each have a polarization angle different from that of the first polarization unit 104A and the second polarization unit 104B.

[0056] The polarization array 104 may include a layer of metal wire gratings. The material of the polarization array 104 may be an opaque material, such as aluminum (Al), gold (Au), silver (Ag), titanium (Ti), niobium (Nb), or a combination of the foregoing. In some embodiments, the metal wire grating may have a film thickness of about 20 nm to about 300 nm. In some embodiments, the metal wire grating may have a period of about 20 nm to about 400 nm. In some embodiments, the metal wire grating may have a fill ratio (or duty cycle) of about 0.2 to about 0.8. The directionality of the grating ridges mainly affects the penetration percentage of polarized light. For example, when the polarized light is parallel to the first polarization unit 104A or the second polarization unit 104B, offset by 45°, or offset by 90°, the intensity of the transmitted light passing through the secondary polarization unit is the maximum value, about 50%, or the minimum value, respectively. Therefore, when the excitation light is polarized light that is offset by 90° from the secondary polarization unit, the secondary polarization unit under the reaction region will block the excitation light from passing through the secondary polarization unit.

[0057] In addition to the polarization angle of the secondary polarization unit, the extinction ratio of the blocking efficiency is also affected by the film thickness of the metal wire, the grating period, the grating profile, and the fill ratio. Peng Li et al. have simulated in "Investigation of achromatic micro polarizer array for polarization imaging in visible-infrared band." Optik, vol 158, April 2018, pp. 1427-1435 that an aluminum wire with a thickness of 160 nm, a fill ratio of 0.5, and a period of 150 nm can achieve an extinction ratio of 10 4 (equivalent to an optical density of 4). In some embodiments, for biosensing applications, an optical density greater than 3 for the light illumination system may be sufficient to block the excitation light.

[0058] The reaction region 108 is disposed on the polarization array 104. The reaction regions 108 may each correspond to one of the first polarization unit 104A or the second polarization unit 104B. In some embodiments, the reaction regions 108 may be formed in the form of nanowells or nanopatterns. Figure 1A In, the reaction region 108 is defined by the openings in the sample isolation layer 106. Therefore, the bottom surface of the reaction region 108 may be the top surface of the polarization array 104. In other embodiments, the sample isolation layer 106 may not have the same as Figure 1AThe opening shown in [figure]. The reaction zone 108 can be formed by modifying a part of the surface of the sample isolation layer 106, such that the modified part can capture a predetermined biological sample. For example, some functional groups on the surface of the sample isolation layer 106 can be modified to capture the desired biological sample.

[0059] In addition, the sample isolation layer 106 where the reaction zone 108 is disposed can be further modified to enhance the immobilization effect of the biological sample. For example, in some embodiments, a self-assembly monolayer (SAM), a functional polymer, or a hydrogel can be used to coat or treat the sample isolation layer 106 to immobilize the biological sample in the reaction zone 108. In other embodiments, the sample isolation layer 106 may not be modified. The biological sample can be immobilized in the reaction zone 108 according to its weight, size, surface charge, or Van der Waals force, etc.

[0060] The sample isolation layer 106 can be formed by sputtering, evaporation, spin-coating, chemical vapor deposition (CVD), molecular beam deposition, any other suitable process, or a combination of the foregoing. For example, chemical vapor deposition can include low-pressure CVD (LPCVD), low-temperature CVD (LTCVD), rapid thermal CVD (RTCVD), plasma enhanced CVD (PECVD), atomic layer deposition (ALD), or a combination of the foregoing.

[0061] According to some embodiments, the material of the sample isolation layer 106 can be transparent, translucent, or opaque. Specifically, in embodiments where the sample isolation layer 106 is transparent, the material of the sample isolation layer 106 can have a light transmittance greater than 85% for light with a wavelength between 400 nm and 750 nm, or preferably greater than 92%. In embodiments where the sample isolation layer 106 is translucent, the material of the sample isolation layer 106 can have a light transmittance greater than 25% and less than 85% for light with a wavelength between 400 nm and 750 nm. In embodiments where the sample isolation layer 106 is opaque, the material of the sample isolation layer 106 can have a light transmittance less than 10% for light with a wavelength between 400 nm and 750 nm, or preferably less than 5%.

[0062] The material of the sample isolation layer 106 may include metals, metal alloys, metal oxides, metal nitrides, silicon, silicon oxide, silicon nitride, or combinations of the foregoing. In some embodiments, the metals, metal alloys, metal oxides, and metal nitrides may include, but are not limited to, silver (Ag), aluminum (Al), gold (Au), niobium (Nb), titanium (Ti), tungsten (W), alloys of the foregoing, titanium oxide (e.g., TiO 2 ), tantalum oxide (e.g., Ta 2 O 5 ), aluminum oxide (e.g., Al 2 O 3 ), niobium oxide (e.g., Nb 2 O 5 ), titanium nitride, tantalum nitride, or combinations of the foregoing.

[0063] Referring again to Figure 1A , the biometric detection system 10A further includes an excitation light source 111. The excitation light source 111 can be used to emit excitation light 112. In some embodiments, the excitation light source 111 may include a plurality of sub-excitation light sources (not shown), and each sub-excitation light source may emit excitation light having a single excitation light wavelength. In some embodiments, the sub-excitation light sources may alternately emit a plurality of excitation lights having various excitation light wavelengths. For example, sub-excitation light sources having different excitation light wavelengths may sequentially emit excitation light. Alternatively, the sub-excitation light sources may emit excitation light in groups. In some embodiments, the excitation light source 111 is a monochromator that continuously emits light from a short wavelength to a long wavelength (or from a long wavelength to a short wavelength). For example, the monochromator may emit light having a wavelength between about 200 nm and about 1000 nm.

[0064] Referring to Figure 1A and Figure 1B , the biometric detection system 10A further includes a front polarizing element 110. The front polarizing element 110 can be used to polarize the excitation light 112 emitted from the excitation light source 111. Accordingly, the biochip 100A can receive the excitation light 112 polarized by the front polarizing element 110. In some embodiments, as shown in Figure 1A and Figure 1B , the front polarizing element 110 may be rotatably orientable such that the front polarizing element 110 can have a polarization angle the same as the first polarization angle of the first polarization unit 104A (as shown in Figure 1B ) or the second polarization angle of the second polarization unit 104B (as shown in Figure 1A ). For example, in Figure 1A , the front polarizing element 110 is rotated to have the second polarization angle, so that the polarized excitation light can pass through the second polarization unit 104B.Figure 1B In this case, the pre-rotation polarization element 110 is rotated to have a first polarization angle, so that the polarized excitation light can pass through the first polarization unit 104A.

[0065] Referring again to Figure 1A , the biological detection system 10A may further include a lens 114 and a photodetector 118. The lens 114 may be any suitable optical lens. After irradiating the biological sample with the polarized excitation light, the lens 114 can focus the emitted light from the biological sample. The photodetector 118 can be used to detect the emitted light. The photodetector 118 may be a photodiode or any suitable light sensing component that can convert the measured light into a current signal. In an embodiment where the photodetector 118 is a photodiode, the photodetector 118 may be connected to the source and drain of a metal-oxide-semiconductor (MOS) transistor (not shown), which can transmit the current to another component, such as another metal-oxide-semiconductor transistor. The another component may include a reset transistor, a current source follower, or a row selector to convert the current into a digital signal, but is not limited thereto.

[0066] In some embodiments, the biological detection system 10A may further include a filter element 116. For example, the filter element 116 may be a rejection filter, which can filter the excitation light and prevent the excitation light from entering the photodetector 118. In some embodiments, the filter element 116 may be disposed on the optical path between the lens 114 and the photodetector 118 as shown in Figure 1A , but the present invention is not limited thereto. In other embodiments, the filter element 116 may also be disposed on the optical path between the biochip 100A and the lens 114. In some embodiments, the filter element 116 may include an absorption filter, an interference filter, a plasmonic metasurface structure, a dielectric metasurface structure, or a combination of the foregoing.

[0067] As described above, according to an embodiment of the present invention, the polarization array of the biochip may include a first polarization unit and a second polarization unit. The first polarization unit has a first polarization angle and the second polarization unit has a second polarization angle. The first polarization angle is different from the second polarization angle. In addition, according to an embodiment of the present invention, the biological detection system may include a front polarizing element, which is rotatably oriented to have the first polarization angle of the first polarization unit or the second polarization angle of the second polarization unit. Thereby, the biological detection system can sequentially detect biological samples corresponding to different types of sub-polarization units, and then can combine fluorescence signals to obtain complete and accurate analysis results. Therefore, in each image, crosstalk between adjacent reaction regions can be reduced, thereby achieving better spatial resolution, and the biochip can be fabricated with a higher array density.

[0068] Next, referring to Figure 1C , Figure 1C is a cross-sectional view of a biological detection system 10B according to other embodiments of the present invention. Figure 1C The biological detection system 10B in Figure 1A is similar to the biological detection system 10A in Figure 1B and

[0069] but the biological detection system 10B may include a biochip 100B, and the biochip 100B further includes a planarization layer 120 disposed between the reaction region 108 and the polarization array 104.

[0070] In some embodiments, the material of the planarization layer 120 may include silicon oxide (SiO 2 ), amorphous silicon (a-Si), aluminum oxide (Al 2 O 3 ), niobium oxide (Nb 2 O 5)), a polymer, or a combination of the foregoing. For example, the polymer may include bisbenzocyclobutene (BCB), polyimide (PI), polymethyl methacrylate (PMMA), cyclic olefin copolymer (COP), polycarbonate (PC), another suitable material, or a combination of the foregoing, but not limited thereto. According to some embodiments, the planarization layer 120 may be transparent or translucent. Specifically, in embodiments where the planarization layer 120 is transparent, the material of the planarization layer 120 may have a light transmittance greater than 85% for light with a wavelength between 400 nm and 750 nm, or preferably greater than 92%. In embodiments where the planarization layer 120 is translucent, the material of the planarization layer 120 may have a light transmittance greater than 25% and less than 85% for light with a wavelength between 400 nm and 750 nm.

[0071] Referring to Figure 1D , Figure 1D is a cross-sectional view of the biochip 200 according to other embodiments of the present invention. The biochip 100A in the above embodiments is an example of an open chamber biochip, and Figure 1D the biochip 200 shown in the embodiment is an example of a flow cell biochip. As Figure 1D shown, the flow cell biochip 200 may include the biochip 100A, a second substrate 122, a spacer layer 124, and a microfluidic channel 126 defined between the first substrate 102 of the biochip 100A and the second substrate 122. The second substrate 122 may be disposed opposite to the first substrate 102 of the biochip 100A. The material of the second substrate 122 may be the same as or similar to the material of the first substrate 102 described above, and will not be repeated here.

[0072] The spacer layer 124 is disposed between the first substrate 102 and the second substrate 122 of the biochip 100A. In some embodiments, the spacer layer 124 may include an adhesive material, such as epoxy acrylate (EA), polyurethane (PU), polyether acrylate, polyesteracrylate (PEA), or a combination of the foregoing. In some embodiments, the spacer layer 124 may further include other materials, such as silicon oxide (SiO 2) Amorphous silicon (a-Si), polymers, or combinations thereof. For example, the polymer may include benzocyclobutene (BCB), polyimide (PI), polymethyl methacrylate (PMMA), cyclic olefin polymer (COP), polycarbonate (PC), another suitable material, or combinations thereof, but not limited thereto.

[0073] According to other embodiments of the present invention, the second substrate 122 and the spacer layer 124 may be formed in the same process such that the second substrate 122 and the spacer layer 124 are integrated into the same structure. In an embodiment where the second substrate 122 and the spacer layer 124 are integrated into the same structure, polydimethylsiloxane (PDMS) may be used to form the second substrate 122 and the spacer layer 124. After forming the second substrate 122 and the spacer layer 124, the second substrate 122 and the spacer layer 124 may be attached to the first substrate 102 by surface arc treatment or surface plasma treatment. In some embodiments, the spacer layer 124 may have a thickness between about 25 μm and about 1000 μm, preferably between about 50 μm and about 500 μm, more preferably between about 50 μm and about 200 μm.

[0074] The microfluidic channel 126 may be defined between the first substrate 102 and the second substrate 122 of the biochip 100A. Figure 1D In, the reaction zone 108 may face the microfluidic channel 126. The opening 128 may be formed through the second substrate 122 and serve as an inlet and outlet for the biological sample, but the present invention is not limited thereto. In other embodiments, the opening may not be formed on the second substrate 122. Instead, the opening may be formed on the side where the biochip 100A is located. Specifically, an opening (not shown) may be formed in an area other than the reaction zone 108 on the biochip 100A. The microfluidic channel 126 may guide the biological sample to the reaction zone 108, and the biological sample may be fixed in the reaction zone 108.

[0075] Next, refer to Figure 1E , Figure 1E is a cross-sectional view of the biological detection system 10C according to other embodiments of the present invention. Figure 1E The biological detection system 10C in is similar to Figure 1A and Figure 1B The biological detection system 10A in, but the biological detection system 10C uses a reflective device to guide the excitation light to the biochip 100A.

[0076] As Figure 1EAs shown, the biological detection system 10C may further include a spectroscopic element 130 as a reflective device, and the spectroscopic element 130 may direct the excitation light to the biochip 100A. In some embodiments, the spectroscopic element 130 may be disposed on the optical path between the lens 114 and the filter element 116. According to some embodiments of the present invention, the spectroscopic element 130 may include a dichroic filter (e.g., an interference filter) that has the property of reflecting short wavelengths less than 580 nm and transmitting long wavelengths greater than 580 nm. The cut-off wavelengths for reflection and transmission may be changed based on the excitation and emission characteristics of the fluorescent molecules. Since the spectra of the polarized excitation light 112' and the emission light 132 from the biological sample are different, the spectroscopic element 130 may reflect the polarized excitation light 112' to the biochip 100A and may transmit the emission light 132. According to some other embodiments, the spectroscopic element 130 may include a beam splitter with a 50 / 50 splitting ratio, but the present invention is not limited thereto. In still other embodiments, the spectroscopic element 130 may also be a beam splitter with a splitting ratio other than 50 / 50, such as 60 / 40 or 70 / 30. In some embodiments, the beam splitter may be a cube beam splitter, a plate beam splitter, or a pellicle beam splitter.

[0077] Figure 1E In [the figure], the front polarizing element 110 is disposed on the optical path between the excitation light source 111 and the spectroscopic element 130, but the present invention is not limited thereto. In other embodiments, the front polarizing element 110 may be disposed on the optical path between the lens 114 and the biochip 100A (not shown).

[0078] Referring to Figure 2A and Figure 2B , Figure 2A and Figure 2B are top views of the biochip 100A according to some embodiments of the present invention. As Figure 2A and Figure 2B shown, the first polarization unit 104A and the second polarization unit 104B may have a rectangular or triangular shape. In other embodiments, the first polarization unit 104A and the second polarization unit 104B may have a hexagonal shape (not shown). It should be noted that Figure 1A and Figure 1B The cross-sectional views are taken along the section line A-A' in Figure 2A or Figure 2B . In addition, for the sake of simplicity, Figure 2A and Figure 2B omit the sample isolation layer 106.

[0079] As Figure 2A or Figure 2BAs shown, at least one first polarization unit 104A can be directly adjacent to the second polarization unit 104B, and at least one first polarization unit 104A can be separated from other first polarization units 104A via the second polarization unit 104B. The term "directly adjacent to" as described herein means that at least one first polarization unit 104A can share an edge with the second polarization unit 104B. With the above configuration, crosstalk between the fluorescence signals corresponding to the first polarization unit 104A and the second polarization unit 104B can be reduced, and thus a higher fluorescence imaging spatial resolution can be achieved.

[0080] Embodiments of the present invention also provide a biological detection method using the biological detection system in the above embodiments. Refer to Figure 3 and Figures 4A to 4B , Figure 3 is a flowchart of a biological detection method 300 according to some embodiments of the present invention, and Figures 4A to 4B is a schematic diagram showing the use of the biological detection system 10A to detect biological samples 400A and 400B according to some embodiments of the present invention. The biological detection method 300 may include the following steps. The biological detection method 300 starts with step 302 to provide the biochip 100A included in the biological detection system 10A.

[0081] Next, refer to Figure 3 step 304 and Figure 4A , the biological samples 400A and 400B are fixed in the reaction area 108. Specifically, an appropriate amount of solution containing the biological sample is applied to the biochip 100A. After the solution containing the biological sample dries, the excessive residual biological sample is removed from the area outside the reaction area 108, and the biological sample can be fixed in each reaction area 108. Figure 4A In

[0082] In some embodiments, the biological samples 400A and 400B may include biomolecules, chemical molecules, or a combination of the foregoing, but are not limited thereto. In some embodiments, the biomolecules may include DNA, RNA, proteins, or a combination of the foregoing, but are not limited thereto. According to some embodiments, the biological samples 400A and 400B can be analyzed to determine gene sequences, DNA-DNA hybridization, single nucleotide polymorphisms (SNP), protein interactions, peptide interactions, antigen-antibody interactions, glucose monitoring, cholesterol monitoring, etc.

[0083] Next, refer toFigure 3 Step 306 and Figure 4A , perform a first detection step to obtain a first fluorescence signal 400AS emitted by the biological sample 400A, where the biological sample 400A is fixed in the reaction region 108 corresponding to the first polarization unit 104A. Specifically, rotate the front polarizing element 110 so that it has the same polarization angle as the first polarization angle of the first polarization unit 104A. Irradiate the biological sample 400A with the polarized excitation light 112' passing through the front polarizing element 110 and the first polarization unit 104A. At the same time, the biological sample 400B is not irradiated by the polarized excitation light 112' because the second polarization unit 104B blocks the polarized excitation light 112', and thus the biological sample 400B does not generate a fluorescence signal, thereby preventing crosstalk. In step 306, only the biological sample 400A emits the first fluorescence signal 400AS. The diverging first fluorescence signal 400AS can be focused by the lens 114 to generate a larger signal intensity, and then the first fluorescence signal 400AS can be detected by the photodetector 118.

[0084] Next, refer to Figure 3 Step 308 and Figure 4B , perform a second detection step to obtain a second fluorescence signal 400BS emitted by the biological sample 400B, where the biological sample 400B is fixed in the reaction region 108 corresponding to the second polarization unit 104B. Specifically, rotate the front polarizing element 110 so that it has the same polarization angle as the second polarization angle of the second polarization unit 104B. Irradiate the biological sample 400B with the polarized excitation light 112' passing through the front polarizing element 110 and the second polarization unit 104B. At the same time, the biological sample 400A is not irradiated by the polarized excitation light 112' because the first polarization unit 104A blocks the polarized excitation light 112', and thus the biological sample 400A does not generate a fluorescence signal, thereby preventing crosstalk. In step 308, only the biological sample 400B emits the second fluorescence signal 400BS. The diverging second fluorescence signal 400BS can be focused by the lens 114 to generate a larger signal intensity, and then the second fluorescence signal 400BS can be detected by the photodetector 118.

[0085] Next, refer to Figure 3In step 310, the first fluorescence signal 400AS and the second fluorescence signal 400BS are combined to obtain the complete fluorescence signal from the biological samples 400A and 400B, thereby completing the process of biological detection. Specifically, two images respectively including the first fluorescence signal 400AS and the second fluorescence signal 400BS can be merged, and then the merged image can be analyzed to obtain the complete fluorescence signal. Alternatively, an image including the first fluorescence signal 400AS and an image including the second fluorescence signal 400BS can be analyzed separately, and then the results of the two images can be combined. The above images can be analyzed by the presence or absence of the fluorescence signal from the biological sample, or the different molecules immobilized in the reaction region 108 can be distinguished by the fluorescence spectrum of the fluorescence signal. Although the order of the first detection step and the second detection step is as described above, it should be understood that the second detection step can be performed before the first detection step.

[0086] In summary, according to some embodiments of the present invention, the polarization array of the biochip may include a first polarization unit and a second polarization unit. The first polarization unit has a first polarization angle and the second polarization unit has a second polarization angle. The first polarization angle is different from the second polarization angle. In addition, according to some embodiments of the present invention, the biological detection system may include a front polarization element, and the front polarization element is rotatably orientable so that it has the same polarization angle as the first polarization angle or the second polarization angle. With the above configuration and the two-step detection method, the crosstalk between the fluorescence signals of adjacent reaction regions can be reduced, and a biochip can be manufactured to have a higher array density.

[0087] Although some embodiments of the present invention and their advantages have been described in detail, it should be understood that various changes, substitutions, and modifications can be made without departing from the spirit and scope of the invention as defined by the protection scope of the present invention. For example, those of ordinary skill in the art to which the present invention pertains should easily understand that many components, functions, processes, and materials described herein can be changed without departing from the scope of the present invention. Furthermore, the scope of the present application is not limited to the specific embodiments of the processes, machines, manufactures, compositions of matter, methods, and steps described in the specification. Those of ordinary skill in the art to which the present invention pertains can easily understand from the present invention that any processes, machines, manufactures, compositions of matter, methods, or steps developed currently or in the future, as long as they can achieve substantially the same functions or achieve substantially the same results as the corresponding embodiments described herein, can be used according to the embodiments of the present invention. Therefore, the protection scope of the present invention includes the above processes, machines, manufactures, compositions of matter, methods, or steps.

Claims

1. A biochip, comprising: a first substrate configured to receive an excitation light from an excitation light source; a polarization array including a plurality of first polarization units and a plurality of second polarization units, the first polarization units having a first polarization angle and the second polarization units having a second polarization angle, the first polarization angle being different from the second polarization angle, wherein at least one first polarization unit is directly adjacent to the second polarization units and the at least one first polarization unit is separated from other first polarization units via the second polarization units, wherein the polarization array has a light input surface and a light output surface, the light input surface of the polarization array being disposed on the first substrate and configured to receive the excitation light that penetrates the first substrate; and a plurality of reaction regions disposed on the light output surface of the polarization array, wherein each of the reaction regions corresponds to one of the first polarization units or the second polarization units.

2. The biochip according to claim 1, wherein, the reaction regions are formed in the form of nanopores or nanoscale patterns, and wherein the first substrate is transparent or translucent.

3. The biochip according to claim 1, wherein, the absolute value difference between the first polarization angle and the second polarization angle is 90°.

4. The biochip according to claim 1, wherein, the first polarization units and the second polarization units have a triangular, rectangular or hexagonal shape.

5. The biochip according to claim 1, further comprising a planarizing layer disposed between the reaction regions and the polarization array.

6. The biochip according to claim 1, wherein, the reaction regions are modified to immobilize a biological sample on the reaction regions.

7. The biochip according to claim 1, further comprising: a second substrate disposed opposite to the first substrate; a spacer layer disposed between the first substrate and the second substrate; and a microfluidic channel defined between the first substrate and the second substrate, wherein the reaction regions face the microfluidic channel, and wherein the microfluidic channel is configured to guide a biological sample to the reaction regions.

8. A biological detection system, comprising: an excitation light source configured to emit an excitation light; a front polarization element configured to polarize the excitation light; the biochip according to claim 1 configured to receive the polarized excitation light, wherein the front polarization element is rotatably orientable such that the front polarization element has a polarization angle that is the same as the first polarization angle or the second polarization angle; a lens configured to focus an emission light from a biological sample immobilized on the reaction regions of the biochip; and a light detector configured to detect the emission light.

9. The biological detection system according to claim 8, further comprising a filter element configured to filter the excitation light entering the light detector.

10. The biological detection system according to claim 8, wherein, the excitation light source includes a plurality of sub-excitation light sources, and wherein each of the sub-excitation light sources emits an excitation light having a single excitation light wavelength.

11. The biological detection system according to claim 8 further includes a beam splitting element for guiding the excitation light to the biochip, wherein the beam splitting element includes a dichroic filter which has a reflection characteristic for short wavelengths and a transmission characteristic for long wavelengths.

12. A biological detection method, comprising: providing the biological detection system according to claim 8; fixing a biological sample to the reaction area of the biochip; performing a first detection step to obtain a first fluorescence signal, which is from the biological sample fixed to the reaction area corresponding to the first polarization unit; performing a second detection step to obtain a second fluorescence signal, which is from the biological sample fixed to the reaction area corresponding to the second polarization unit; and combining the first fluorescence signal and the second fluorescence signal.

13. The biological detection method according to claim 12, wherein the first detection step includes: rotating the front polarizing element so that the front polarizing element has a polarization angle the same as the first polarization angle; and irradiating the biological sample fixed to the reaction area corresponding to the first polarization unit with an excitation light, wherein the excitation light passes through the front polarizing element and the first polarization unit.

14. The biological detection method according to claim 12, wherein the second detection step includes: rotating the front polarizing element so that the front polarizing element has a polarization angle the same as the second polarization angle; and irradiating the biological sample fixed to the reaction area corresponding to the second polarization unit with an excitation light, wherein the excitation light passes through the front polarizing element and the second polarization unit.

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