Imaging method for detecting spectral displacement and biosensor

Abstract

The invention provides an imaging method for detecting spectral displacement and a biosensor. The imaging method comprises the following steps: receiving a first light beam through a photodiode array to simultaneously obtain a first image signal which has first image intensity and corresponds to a first optical filter and a second image signal which has second image intensity and corresponds to a double-layer optical filter comprising a second optical filter and a third optical filter; receiving the second light beam through a photodiode array to simultaneously obtain a third image signal which has third image intensity and corresponds to the first optical filter and a fourth image signal which has fourth image intensity and corresponds to the double-layer optical filter; the spectral displacement from the first light beam to the second light beam is determined by comparing the first image intensity and the second image intensity with the third image intensity and the fourth image intensity. Because the image signals corresponding to different optical filters are obtained at the same time, the signal-to-noise ratio and the elastic degree of spectral displacement detection can be improved, and the immediate detection of spectral displacement and the miniaturization of the biological sensor are realized.

Description

Technical Field

[0001] The present disclosure relates to imaging methods and biosensors for detecting spectral shifts, and more particularly to imaging methods and biosensors for detecting spectral shifts using multiple filters. Background Art

[0002] After the target biomolecule reacts with the receptor on the substrate of the biosensor, the surface property change of the substrate can be measured based on the angle or wavelength of the detection light. For example, when the refractive index of the substrate changes, the resonance spectrum of the detection light may show a red shift or a blue shift accordingly. However, if the resonance spectrum has a high quality factor (Q factor), it will be difficult to detect the spectral shift in real time. Summary of the invention

[0003] The present disclosure provides an imaging method for detecting spectral shift. The imaging method includes receiving a first light beam through a first photodiode and a second photodiode to simultaneously obtain a first image signal with a first image intensity from the first photodiode, and a second image signal with a second image intensity from the second photodiode. The first image signal corresponds to the first light beam filtered by the first filter, and the second image signal corresponds to the first light beam filtered by the first double-layer filter. The first double-layer filter includes a second filter superimposed on a third filter, wherein the third filter has the same wavelength range as the first filter. The difference between the peak wavelength of the first image signal and the central wavelength of the first filter is within plus or minus 5 nanometers. The difference between the peak wavelength of the first image signal and the start / cutoff wavelength of the second filter is within plus or minus 5 nanometers. The imaging method further includes receiving a second light beam through the first photodiode and the second photodiode to simultaneously obtain a third image signal with a third image intensity from the first photodiode, and a fourth image signal with a fourth image intensity from the second photodiode. The third image signal corresponds to the second light beam filtered by the first filter, and the fourth image signal corresponds to the second light beam filtered by the first double-layer filter. The imaging method further includes comparing the difference between the first image intensity and the second image intensity and the third image intensity and the fourth image intensity to determine the spectral shift from the first light beam to the second light beam.

[0004] In some embodiments, when the first light beam is received by the first photodiode and the second photodiode, the second image intensity is half of the first image intensity.

[0005] In some embodiments, a slope of the transition band of the first filter and a slope of the transition band of the second filter are steeper than a slope of the first image signal.

[0006] In some embodiments, when the first light beam is received by the first photodiode and the second photodiode, the wavelength range of the first image signal completely falls within the wavelength range of the first filter.

[0007] In some embodiments, the first filter is a short bandpass filter and the second filter is a long wave pass filter or a long bandpass filter.

[0008] In some embodiments, the short bandpass filter has a bandwidth in the range of 20 nanometers to 60 nanometers.

[0009] In some embodiments, determining the spectral shift from the first light beam to the second light beam comprises:

[0010]

[0011] Get the signal value γ1 and signal value γ2, where R is the balance parameter, for the signal value γ1, I S and I A is the first image intensity and the second image intensity, for the signal value γ2, I S and I A are the third image intensity and the fourth image intensity. The spectral shift is determined by the difference between the signal value γ1 and the signal value γ2.

[0012] In some embodiments, when the first light beam is received by the first photodiode and the second photodiode, the signal value γ1 is equal to 0.

[0013] In some embodiments, the first filter is a long bandpass filter and the second filter is a short wave pass filter or a short bandpass filter.

[0014] In some embodiments, determining the spectral shift from the first light beam to the second light beam comprises:

[0015]

[0016] Get the signal value γ1 and signal value γ2, where R is the balance parameter, for the signal value γ1, I L and I B is the first image intensity and the second image intensity, for the signal value γ2, I L and I B are the third image intensity and the fourth image intensity. The spectral shift is determined by the difference between the signal value γ1 and the signal value γ2.

[0017] In some embodiments, receiving the first light beam through the first photodiode and the second photodiode further includes receiving the first light beam through the third photodiode to simultaneously obtain a fifth image signal having a fifth image intensity from the third photodiode, the fifth image signal corresponding to the first light beam filtered by the fourth filter, wherein the fourth filter has the same wavelength range as the second filter. Receiving the second light beam through the first photodiode and the second photodiode further includes receiving the second light beam through the third photodiode to simultaneously obtain a sixth image signal having a sixth image intensity from the third photodiode, the sixth image signal corresponding to the second light beam filtered by the fourth filter.

[0018] In some embodiments, the first filter is a short bandpass filter and the second filter is a long bandpass filter. In such embodiments, determining the spectral shift of the first light beam to the second light beam comprises determining the spectral shift of the first light beam to the second light beam by:

[0019]

[0020] or

[0021]

[0022] The signal value γ1 and the signal value γ2 are obtained, where R is the balance parameter. For the signal value γ1 (I S , I A , I L ) are the first image intensity, the second image intensity and the fifth image intensity. For the signal value γ2, (I S , I A , I L ) are the third image intensity, the fourth image intensity and the sixth image intensity. The spectral shift is determined by the difference between the signal value γ1 and the signal value γ2.

[0023] In some embodiments, when I L -RI A =0, formula (III) is simplified to

[0024]

[0025] When I S -RI A =0, formula (IV) is simplified to

[0026]

[0027] In some embodiments, the first filter is a long bandpass filter and the second filter is a short bandpass filter. In such embodiments, determining the spectral shift of the first light beam to the second light beam comprises determining the spectral shift of the first light beam to the second light beam by:

[0028]

[0029] or

[0030]

[0031] The signal value γ1 and the signal value γ2 are obtained, where R is the balance parameter. For the signal value γ1 (I L , I B , I S ) are the first image intensity, the second image intensity and the fifth image intensity. For the signal value γ2, (I L , I B , I S ) are the third image intensity, the fourth image intensity and the sixth image intensity. The spectral shift is determined by the difference between the signal value γ1 and the signal value γ2.

[0032] In some embodiments, when I S -RI B =0, formula (VII) is simplified to

[0033]

[0034] When I L -RI B =0, formula (VIII) is simplified to

[0035]

[0036] In some embodiments, receiving the first light beam through the first photodiode and the second photodiode further includes receiving the first light beam through the third photodiode to simultaneously obtain a fifth image signal having a fifth image intensity from the third photodiode, the fifth image signal corresponding to the first light beam filtered by the second double-layer filter, the second double-layer filter including a fourth filter superimposed on the fifth filter, wherein the fifth filter has the same wavelength range as the first filter. Receiving the second light beam through the first photodiode and the second photodiode further includes receiving the second light beam through the third photodiode to simultaneously obtain a sixth image signal having a sixth image intensity from the third photodiode, the sixth image signal corresponding to the second light beam filtered by the second double-layer filter.

[0037] In some embodiments, the first filter is a central bandpass filter, the second filter is a shortwavepass filter or a short bandpass filter, and the fourth filter is a longwavepass filter or a long bandpass filter.

[0038] In some embodiments, determining the spectral shift from the first light beam to the second light beam comprises:

[0039]

[0040] The signal value γ1 and the signal value γ2 are obtained, where R is the balance parameter. For the signal value γ1 (I C , I B , I A ) are the first image intensity, the second image intensity and the fifth image intensity. For the signal value γ2, (I C , I B , I A ) are the third image intensity, the fourth image intensity and the sixth image intensity. The spectral shift is determined by the difference between the signal value γ1 and the signal value γ2.

[0041] The present disclosure provides a biosensor for performing an imaging method for detecting spectral shift. The biosensor includes a photodiode array and a filter array above the photodiode array, the photodiode array includes a first photodiode and a second photodiode, the filter array includes a first filter and a first double-layer filter, wherein the first photodiode and the second photodiode correspond to the first filter and the first double-layer filter, respectively. The biosensor further includes a sample substrate above the filter array and a light source above the sample substrate.

[0042] In some embodiments, the light source is configured to produce a surface plasmon resonance spectrum.

[0043] In some embodiments, the wavelength range of the light source covers the wavelength range of the filter array.

[0044] According to an embodiment of the present disclosure, an imaging method for detecting spectral shift includes receiving a light beam to simultaneously obtain multiple image signals and using the obtained image signals to determine the spectral shift, wherein a single-layer filter and a double-layer filter in a filter array of a biosensor respectively filter two of the multiple image signals. Since image signals corresponding to different filters are obtained simultaneously, the signal-to-noise ratio can be increased, and the spectral shift can be detected in real time. A filter array composed of different filters can also miniaturize the biosensor and increase the flexibility of the spectral shift detection range. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] When read in conjunction with the accompanying drawings, various aspects of the present disclosure can be best understood from the following detailed description. It should be noted that, in accordance with standard methods in the industry, various features are not drawn to scale. In fact, the size of various features may be arbitrarily increased or reduced for clarity of discussion.

[0046] Figure 1 According to some embodiments of the present disclosure, a flow chart of an imaging method for detecting spectral shift is shown.

[0047] Figure 2A According to some embodiments of the present disclosure, execution Figure 1Schematic diagram of the biosensor imaging method.

[0048] Figure 2B According to an embodiment of the present disclosure, Figure 2A A partial 3D view of a biosensor.

[0049] Figures 3A to 3C , Figures 4A to 4C , Figures 5A to 5C , Figures 6A to 6C and Figures 7A to 7C According to some embodiments of the present disclosure, Figure 1 Imaging methods and Figures 2A to 2B The spectrum of the biosensor.

[0050] Figure 8 According to some embodiments of the present disclosure, a flow chart of an imaging method for detecting spectral shift is shown.

[0051] Figure 9A According to some embodiments of the present disclosure, execution Figure 8 Schematic diagram of the biosensor imaging method.

[0052] Figures 9B to 9D According to some embodiments of the present disclosure, Figure 9A A partial 3D view of a biosensor.

[0053] Figures 10A to 10C and Figures 11A to 11C According to some embodiments of the present disclosure, Figure 8 Imaging methods and Figures 9A to 9B The spectrum of the biosensor.

[0054] Figure 12A According to some embodiments of the present disclosure, execution Figure 8 Schematic diagram of the biosensor imaging method.

[0055] Figure 12B According to an embodiment of the present disclosure, Figure 12A A partial 3D view of a biosensor.

[0056] Figures 13A to 13C and Figures 14A to 14C According to some embodiments of the present disclosure, Figure 8 Imaging methods and Figures 12A to 12B The spectrum of the biosensor.

[0057] The reference numerals are described as follows:

[0058] 10, 20, 30: Biosensors

[0059] 10a, 10b, 10c, 10c′, 20a, 20b, 20c, 20c′, 20c″, 30a, 30b, 30c, 30c′, 30c″: beam

[0060] 100: Photodiode array

[0061] 110: Photodiode

[0062] 200: Filter array

[0063] 210: First filter

[0064] 210a: Arrow

[0065] 210t: Transition zone

[0066] 212: The third filter

[0067] 214: Fifth filter

[0068] 220: Second filter

[0069] 220a: Arrow

[0070] 220t: Transition zone

[0071] 222: The fourth filter

[0072] 230: Double-layer filter

[0073] 240: Bottom

[0074] 250: Fourth filter

[0075] 260: Double-layer filter

[0076] 300: Sample substrate

[0077] 310, 312, 314, 320, 322, 324, 330, 332, 334, 340, 342, 344, 350, 352, 354, 360, 362, 364, 370, 372, 374, 380, 382, ​​384, 390, 392, 394: Spectrum

[0078] 310a: Arrow

[0079] 400: Light source

[0080] AA: Line

[0081] S100, S200: Method

[0082] S102, S104, S106, S202, S204, S206: Steps DETAILED DESCRIPTION

[0083] In order to realize the different features of the mentioned subject matter, the following disclosure provides many different embodiments or examples. Specific examples of components, configurations, etc. are described below to simplify the present disclosure. Of course, these are merely examples and are not restrictive. For example, in the following description, forming a first feature on or above a second feature may include an embodiment in which the first feature and the second feature are formed in direct contact, and may also include an embodiment in which an additional feature is formed between the first feature and the second feature so that the first feature and the second feature may not be in direct contact. In addition, the present disclosure may repeat reference numbers and / or letters in various examples. This repetition is for the purpose of simplicity and clarity, and does not itself represent the relationship between the various embodiments and / or configurations discussed.

[0084] Furthermore, spatially relative terms, such as "below," "beneath," "lower," "above," "upper," and the like, may be used herein to facilitate describing one element or feature's relationship to another element or feature as illustrated in the figures. The spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. The device may be otherwise oriented (rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein interpreted accordingly.

[0085] It should be understood that although the terms "first", "second", "third", etc. may be used herein to describe various elements, components, regions, layers and / or parts, these elements, components, regions, layers and / or parts should not be limited by these terms. These terms are only used to distinguish one element, component, region, layer or part from another element, component, region, layer or part. Therefore, the first element, component, region, layer or part discussed below may be referred to as a second element, component, region, layer or part without departing from the teachings of this article.

[0086] The present disclosure provides an imaging method for detecting spectral shift and a biosensor for performing the imaging method. The imaging method includes receiving a light beam to simultaneously obtain multiple image signals, and using the obtained image signals to determine the spectral shift, wherein a single-layer filter and a double-layer filter in a filter array of the biosensor filter two of the image signals respectively. Since image signals corresponding to different filters are obtained at the same time, the signal-to-noise ratio can be increased, and the spectral shift can be detected in real time. In addition, a filter array composed of different filters can miniaturize the biosensor and increase the flexibility of the spectral shift detection range.

[0087] According to some embodiments of the present disclosure, Figure 1 A flow chart of an imaging method S100 for detecting spectral shift is shown, and Figure 2A Draw for execution Figure 1Schematic diagram of a biosensor 10 of an imaging method S100. Specifically, the biosensor 10 includes a photodiode array 100, a filter array 200 above the photodiode array 100, a sample substrate 300 above the filter array 200, and a light source 400 above the sample substrate 300. The light source 400 is configured to emit a light beam 10a toward the sample substrate 300. The light beam 10a passes through the sample substrate 300 or is reflected by the sample substrate 300 to generate a light beam 10b.

[0088] When a biological sample is placed on the sample substrate 300 or reacts to another state on the sample substrate 300, the biological sample may affect the characteristics of the light beam 10b. For example, the light source 400 can be configured to generate a surface plasmon resonance (SPR) spectrum. When the biological sample is placed on the sample substrate 300, the surface plasmon resonance spectrum can show a red shift or a blue shift. In other words, different biological samples on the sample substrate 300 can provide light beams 10b of different wavelengths.

[0089] Next, the light beam 10b is directed to the filter array 200 including a plurality of filters having different wavelength ranges. Specifically, the filter array 200 includes at least one single-layer first filter 210 having a first wavelength range, thereby filtering the light beam 10b into a light beam 10c within the first wavelength range. The filter array 200 also includes at least one double-layer filter 230, wherein the double-layer filter 230 includes a third filter 212 and a second filter 220 overlapping the third filter 212. The third filter 212 has the same first wavelength range as the first filter 210, while the second filter 220 has a second wavelength range that is different from the first wavelength range and partially overlaps with the first wavelength range. Therefore, the double-layer filter 230 filters the light beam 10b into a light beam 10c′ within the overlapping range of the first wavelength range and the second wavelength range.

[0090] In some embodiments, the first filter 210 and the double-layer filter 230 can be arranged alternately in the filter array 200 to form Figure 2A In the embodiment, the first filter 210 and the double filter 230 are adjacent to each other in the filter array 200. When the light beam 10b reaches the filter array 200, the first filter 210 and the double filter 230 simultaneously filter the light beam 10b into the light beam 10c and the light beam 10c′. In such an embodiment, the wavelength range of the light source 400 can cover the wavelength range of the filter array 200 to simultaneously generate both the light beam 10c and the light beam 10c′. For example, the light source 400 can be a halogen lamp or a white light emitting diode (LED) with a wavelength range of 380 nanometers to 1300 nanometers.

[0091] In this document, the terms "single layer" and "double layer" represent the different number of layers between the separate first filter 210 and the overlapping second filter 220 / third filter 212, and are not intended to limit the actual number of layers of the filter. For example, in some embodiments, the separate first filter and the overlapping second filter / third filter can be formed on an additional common substrate of the filter array. It should be noted that unless otherwise specified, the terms "first filter" and "second filter" themselves represent a single layer filter. In addition, Figure 2A Two first optical filters 210 and two double-layer optical filters 230 are depicted as an example, but the optical filter array 200 including a different number of first optical filters 210 and double-layer optical filters 230 is also within the scope of the present disclosure.

[0092] Next, the light beam 10c and the light beam 10c′ filtered by the filter array 200 are directed to the photodiode array 100 including a plurality of photodiodes 110. Each photodiode 110 corresponds to one of the first filter 210 or the double-layer filter 230, and is used to receive one of the light beam 10c or the light beam 10c′. Since the light beam 10c and the light beam 10c′ are filtered by the first filter 210 and the double-layer filter 230, respectively, the photodiode array 100 can provide spectral intensities in two different wavelength ranges through the plurality of photodiodes 110.

[0093] In order to clearly explain the relationship between the photodiode array 100 and the filter array 200, Figure 2B According to an embodiment of the present disclosure, Figure 2A A partial 3D view of a biosensor. Figure 2B As shown in FIG. 1 , the photodiodes 110 are arranged along two directions to form the photodiode array 100. The first optical filter 210 and the double-layer optical filter 230 are arranged along the same two directions, so that each photodiode 110 is disposed under one of the first optical filter 210 or the double-layer optical filter 230. Since each photodiode 110 is aligned with one of the first optical filter 210 or the double-layer optical filter 230, each photodiode 110 is configured to receive a light beam only from one of the first optical filter 210 or the double-layer optical filter 230.

[0094] exist Figure 2A In the embodiment of the present invention, the elements in the biosensor 10 are arranged in a straight line, and the biosensor 10 can be miniaturized. In some other embodiments, as long as the light beam 10b can pass through the filter array 200 and reach the photodiode array 100, the spatial configuration of the photodiode array 100, the filter array 200, the sample substrate 300 and the light source 400 can be non-linear.

[0095] Can be used Figures 2A to 2BThe biosensor 10 performs Figure 1 An imaging method S100 for detecting spectral shift is provided in the present invention. Specifically, step S102 of the imaging method S100 is first performed with a biosensor 10 in an initial state, such as a biosensor 10 without a biological sample or a biosensor 10 with an unreacted biological sample. The light source 400 of the biosensor 10 in the initial state provides a light beam 10a to a sample substrate 300, thereby generating an initial light beam 10b. The initial light beam 10b is filtered by the filter array 200 into an initial light beam 10c and an initial light beam 10c′. The photodiode array 100 simultaneously receives the initial light beam 10c and the initial light beam 10c′, thereby generating a first image signal corresponding to the initial light beam 10c and a second image signal corresponding to the initial light beam 10c′.

[0096] After the biological sample is placed in or reacted in the biosensor 10, the biosensor 10 may perform step S104 of the imaging method S100. Since the biological sample affects the characteristics of the light beam 10b, the later light beam 10b emitted by the sample substrate 300 may have a spectrum different from the initial light beam 10b. Therefore, the spectra of the initial light beam 10c and the initial light beam 10c' are correspondingly shifted into the spectra of the later light beam 10c and the later light beam 10c'. As the photodiode array 100 simultaneously receives the later light beam 10c and the later light beam 10c', the biosensor 10 generates a third image signal corresponding to the later light beam 10c and a fourth image signal corresponding to the later light beam 10c'.

[0097] After obtaining the first to fourth image signals, step S106 of the imaging method S100 is performed to determine the spectral shift caused by the biological sample. By comparing the differences between the first and second image signals and the third and fourth image signals, the spectral shift from the initial light beam 10c and the initial light beam 10c′ to the later light beam 10c and the later light beam 10c′ can be determined, which will be further described in detail below.

[0098] According to one embodiment of the present disclosure, Figures 3A to 3C Draw the corresponding Figure 1 Imaging methods S100 and Figures 2A to 2B The spectrum of the biosensor 10. Figures 3A to 3C As shown in FIG. 1 , the first filter 210 and the third filter 212 are the same short band-pass filters, while the second filter 220 is a long-pass filter having a wavelength range partially overlapping with that of the first filter 210 . Figures 3A to 3CThe dotted area in represents the overlapping wavelength range, i.e., the wavelength range of the double-layer filter 230. For example, the first filter 210 may be a short bandpass filter with a bandwidth of about 40 nanometers, and the overlapping wavelength range of the first filter 210 and the second filter 220 has a bandwidth of about 20 nanometers. In some preferred embodiments, the bandwidth of the bandpass filter may be in the range of 20 nanometers to 60 nanometers.

[0099] Figure 3A The spectrum 310 obtained by the biosensor in the initial state (e.g., step S102) is shown, wherein the spectrum 310 represents the unfiltered light beam from the sample substrate. As described above, the spectrum 310 filtered by the first filter 210 is the first image signal, and the spectrum 310 filtered by the double-layer filter 230 is the second image signal. Figure 3A As shown in , the wavelength range of the spectrum 310 may completely fall within the wavelength range of the first filter 210. In other words, the shortest wavelength of the spectrum 310 may be greater than or equal to the shortest wavelength that can pass through the first filter 210, and the longest wavelength of the spectrum 310 may be less than or equal to the longest wavelength that can pass through the first filter 210. In addition, the boundary of the dot area representing the double-layer filter 230 may bisect the spectrum 310. Therefore, the first image signal may be the complete spectrum 310, and the second image signal is the right half of the spectrum 310.

[0100] Since the first image signal and the second image signal correspond to different parts of the spectrum 310, the first image signal and the second image signal have different image intensities. In this document, the term "image intensity" refers to the area under the spectrum. In other words, the first image intensity of the first image signal is the area under the complete spectrum 310, while the second image intensity of the second image signal is the area under the right half of the spectrum 310.

[0101] The signal value γ1 representing the position of the spectrum 310 can be obtained by using the first image intensity and the second image intensity. Specifically, the signal value γ1 can be obtained by the following formula (I), where R is a balance parameter, I S is the first image intensity, I A The balance parameter R is used to reduce the intensity impact caused by the difference in the number of layers between the single-layer first optical filter 210 and the double-layer optical filter 230 .

[0102]

[0103] For the biosensor in the initial state, the second image intensity can be half of the first image intensity, so that the signal value γ1 is equal to 0 to simplify the determination of the spectrum shift. Specifically, the difference between the peak wavelength of the first image signal and the central wavelength of the first filter 210 can be within plus or minus 5 nanometers, and the difference between the peak wavelength of the first image signal and the cut-on wavelength of the second filter 220 can also be within plus or minus 5 nanometers, so that the central line of the spectrum 310 is substantially aligned with the central wavelength of the first filter 210 and the cut-on wavelength of the double-layer filter 230 (refer to Figure 3A Line AA in the figure).

[0104] In addition to the difference between the specific wavelengths, the slope of the filter's transition band (i.e., the wavelength range between the passband and the stopband) is steeper than the slope of the image signal, which also contributes to the signal value γ1 being equal to 0. In other words, as the spectral intensity decreases along the filter's transition band and the spectral curve of the image signal, the wavelength range spanned by the filter is shorter than the wavelength range spanned by the image signal. Figure 3A For example, the slope of the transition band 210t of the first filter 210 ( Figure 3A The slope of the transition band 220t of the second filter 220 (indicated by the arrow 210a in FIG. Figure 3A The slope of the first image signal (ie, the complete spectrum 310) is greater than that of the first image signal (ie, the complete spectrum 310). Figure 3A Therefore, the spectrum boundary of the double-layer filter 230 is more likely to bisect the spectrum 310 and generate a signal value γ1 equal to zero.

[0105] Figure 3B The spectrum 312 obtained by the biosensor in the later state (e.g., step S104) is shown, where the spectrum 312 represents the unfiltered light beam from the sample substrate. As described above, the spectrum 312 filtered by the first filter 210 is a third image signal with a third image intensity, and the spectrum 312 filtered by the double filter 230 is a fourth image signal with a fourth image intensity. The signal value γ2 representing the position of the spectrum 312 can be obtained by formula (I), where R is the balance parameter, I S is the third image intensity, I A is the fourth image intensity. Figure 3B As shown in , the fourth image intensity is twice greater than the third image intensity, resulting in a signal value γ2 greater than 0.

[0106] Since the signal value γ1 and the signal value γ2 represent the positions of the spectrum 310 and the spectrum 312, respectively, the spectral shift from the spectrum 310 to the spectrum 312 can be determined by the difference between the signal value γ1 and the signal value γ2. In other words, by comparing the difference between the first image intensity and the second image intensity and the third image intensity and the fourth image intensity, the spectral shift from the biosensor beam in the initial state to the biosensor beam in the later state can be determined. Figure 3A and Figure 3B , when the spectrum 310 with a shorter wavelength is shifted to the spectrum 312 with a longer wavelength, the signal value γ1 is correspondingly changed to the signal value γ2. This means that when the spectrum 310 is red-shifted to the spectrum 312, the signal value γ changes from 0 to a positive number.

[0107] Similarly, the change in the signal value γ can also determine the spectral blue shift. Figure 3C The spectrum 314 obtained by the biosensor in the later state is shown, where the spectrum 314 represents the unfiltered light beam from the sample substrate. Figure 3A and Figure 3C When corresponding to step S102 and step S104, the spectrum 314 filtered by the first filter 210 is a third image signal with a third image intensity, and the spectrum 312 filtered by the double-layer filter 230 is a fourth image signal with a fourth image intensity. The signal value γ2 representing the position of the spectrum 314 can be obtained by formula (I), so that the signal value γ2 is less than 0. Therefore, when the spectrum 310 is blue-shifted to the spectrum 314, the signal value γ changes from 0 to a negative number.

[0108] In addition to determining the redshift and blueshift of the spectrum, if the signal value γ2 remains the same as the signal value γ1, the signal value γ1 and the signal value γ2 can be used to determine that the spectrum has not shifted. For example, Figure 3A Corresponding to the biosensor in the initial state, the signal value γ1 is equal to 0. If the biological sample in the biosensor remains unreacted, the spectrum of the light beam from the sample substrate can remain unchanged. Therefore, the spectrum without red shift and blue shift causes the signal value γ to remain 0.

[0109] use Figures 2A to 2B The biosensor 10 performs Figure 1The imaging method S100 can bring advantages. The filter array 200 includes a first filter 210 and a double-layer filter 230 of different wavelength ranges, so that the photodiode array 100 can simultaneously receive the light beam 10c and the light beam 10c′. Therefore, two image signals corresponding to the two filters (such as the first image signal and the second image signal in step S102) can be obtained at the same time to generate a signal value γ of the spectrum. Measuring two image signals at one time reduces the impact of noise on the signal value γ, thereby improving the accuracy of spectral shift determination. Measuring two image signals at one time can also output the signal value γ immediately, so even if the spectrum has a high quality factor, the spectral shift can be detected instantly.

[0110] exist Figures 3A to 3C In the illustrated embodiment, spectrum 310 to spectrum 314 have peak values. According to another embodiment of the present disclosure, Figures 4A to 4C Draw the corresponding Figure 1 Imaging methods S100 and Figures 2A to 2B Spectra 320 to 324 of the biosensor. Figures 4A to 4C The steps and biosensors are similar to Figures 3A to 3C ,but Figures 4A to 4C The biological sample of FIG. 100 causes the spectra 320 to 324 to have valley values. The imaging method S100 may also determine the spectrum shift of the spectra 320 to 324 , but the signal value results of the spectra 320 to 324 are different from those of the spectra 310 to 314 .

[0111] refer to Figure 4A , for the biosensor in the initial state, the difference between the peak wavelength of the spectrum 320 and the central wavelength of the first filter 210 can be within plus or minus 5 nanometers, and the difference between the peak wavelength of the spectrum 320 and the starting wavelength of the second filter 220 can be within plus or minus 5 nanometers. Since the image intensity filtered by the double-layer filter 230 is half of the image intensity filtered by the first filter 210, the signal value γ equal to 0 can be obtained by formula (I). When the spectrum 320 is shifted to the spectrum 322 of Figure 4B, the image intensity filtered by the double-layer filter 230 is twice less than the image intensity filtered by the first filter 210. In addition, when the spectrum 320 is shifted to Figure 4C When the spectrum 324 is obtained, the image intensity filtered by the double filter 230 is twice greater than the image intensity filtered by the first filter 210. Therefore, the signal value γ changes from 0 to a negative number when the spectrum is red-shifted, and the signal value γ changes from 0 to a positive number when the spectrum is blue-shifted. Figures 3A to 4C The results of detecting the spectral shift of the embodiment are listed in Table 1 below.

[0112] Table 1

[0113] Spectrum with peak value Spectrum with valley value Initial state or no displacement γ = 0 γ = 0 Red shift γ > 0 γ<0 Blue shift γ<0 γ > 0

[0114] As mentioned above, in Figures 3A to 3C In the embodiment of the present invention, the first filter 210 and the second filter 220 are respectively a short-band pass filter and a long-wave pass filter. In some other embodiments, the first filter 210 and the second filter 220 can use other filter combinations according to the spectrum of the light beam from the sample substrate. For example, Figures 5A to 7C According to some embodiments of the present disclosure, corresponding Figure 1 Imaging methods S100 and Figures 2A to 2B Spectrum of the biosensor 10. Figures 5A to 7C The steps and biosensors are similar to Figures 3A to 3C , but changes with spectrum 330 to spectrum 354 Figures 5A to 7C Filter.

[0115] refer to Figures 5A to 5C , the first filter 210 and the third filter 212 are the same short band-pass filters, and the second filter 220 is a long band-pass filter that partially overlaps the wavelength range of the first filter 210. Since the wavelength range of the first filter 210, the wavelength range of the double-layer filter 230, and the position of the spectrum 330 are similar Figures 3A to 3C , the signal value γ of spectrum 330 to spectrum 334 can be obtained by formula (I), where R is the equilibrium parameter, I S is the intensity of the image filtered by the first filter 210, I A is the intensity of the image filtered by the double-layer filter 230.

[0116] refer to Figures 6A to 6C , the first filter 210 and the third filter 212 are the same long-bandpass filters, and the second filter 220 is a short-pass filter that partially overlaps the wavelength range of the first filter 210. For the spectrum 340, the first image signal is the complete spectrum 340, and the second image signal is the left half of the spectrum 340. In order to obtain a positive signal value γ when the spectrum 340 is red-shifted to the spectrum 342, and to obtain a negative signal value γ when the spectrum 340 is blue-shifted to the spectrum 344, the signal value γ can be obtained by the following formula (II), where R is a balance parameter, I L is the intensity of the image filtered by the first filter 210, I B is the intensity of the image filtered by the double-layer filter 230.

[0117]

[0118] For spectrum 340, the difference between the peak wavelength of spectrum 340 and the central wavelength of the first filter 210 is within plus or minus 5 nanometers, and the difference between the peak wavelength of spectrum 340 and the cut-off wavelength of the second filter 220 can be within plus or minus 5 nanometers, so that the image intensity filtered by the double-layer filter 230 (i.e., the second image intensity) is half of the image intensity filtered by the first filter 210 (i.e., the first image intensity), and the signal value γ1 equal to 0 can be obtained by formula (II). When spectrum 340 shifts to spectrum 342, the spectrum red shift causes the signal value γ1 to become a positive signal value γ2. When spectrum 340 shifts to spectrum 344, the spectrum blue shift causes the signal value γ1 to become a negative signal value γ2.

[0119] refer to Figures 7A to 7C , the first filter 210 and the third filter 212 are the same long-bandpass filters, and the second filter 220 is a short-bandpass filter that partially overlaps the wavelength range of the first filter 210. Since the wavelength range of the first filter 210, the wavelength range of the double-layer filter 230, and the position of the spectrum 350 are similar Figures 6A to 6C , the signal value γ of spectrum 350 to spectrum 354 can be obtained by formula (II), where R is the equilibrium parameter, I L is the intensity of the image filtered by the first filter 210, I B is the intensity of the image filtered by the double-layer filter 230.

[0120] exist Figure 1 and Figures 2A to 2B In the illustrated embodiment, the filter array 200 filters the light beam 10b into the light beam 10c and the light beam 10c′, so that the biosensor 10 simultaneously provides two image signals filtered by the first filter 210 and the double-layer filter 230. In some other embodiments, the filter array can filter the light beam from the sample substrate into more than two light beams with different spectra, thereby simultaneously providing more than two image signals to determine the spectral shift. Such an adjusted filter array can amplify the detection range of the spectral shift. In other words, the filter array can include different filter combinations to match the wavelength and spectral shift of the light beam from the biological sample. The filter combination of the filter array can not only miniaturize and simplify the biosensor, but also increase the flexibility of the spectral shift detection range of the biosensor.

[0121] According to some embodiments of the present disclosure, Figure 8 A flow chart of an imaging method S200 for detecting spectral shift is shown. Figure 9A Draw for execution Figure 8 A schematic diagram of a biosensor 20 of an imaging method S200 is shown. Figures 9B to 9D Draw Figure 9AA partial three-dimensional view of the biosensor 20. The biosensor 20 is similar to Figures 2A to 2B The biosensor 10 is a biosensor 10, but the filter array 200 of the biosensor 20 includes at least one single-layer first filter 210 having a first wavelength range, at least one double-layer filter 230 having an overlapping range of the first wavelength range and the second wavelength range, and at least one single-layer fourth filter 222 having the same second wavelength range as the second filter 220.

[0122] In some embodiments, the first optical filter 210 and the third optical filter 212 may be located in a lower layer of the optical filter array 200, while the second optical filter 220 and the fourth optical filter 222 may be located in an upper layer of the optical filter array 200, thereby simplifying the process of manufacturing the optical filter array 200. In such an embodiment, the fourth optical filter 222 may be disposed on a bottom layer 240, wherein the bottom layer 240 is located in the same lower layer as the first optical filter 210. The bottom layer 240 may be formed of a material having a high transmittance to the light beam 20b, so that the fourth optical filter 222 on the bottom layer 240 may be regarded as a single-layer fourth optical filter 222.

[0123] in addition, Figures 9B to 9D The first filter 210, the fourth filter 222 and the double-layer filter 230 are shown in different proportions and configurations. Figures 9B to 9D As shown in , the three filters can be arranged alternately, or the same filters can be arranged adjacent to each other. Figures 9B to 9D The depicted embodiment is provided as an example, and filter arrays 200 including different numbers and configurations of filters are also within the scope of the present disclosure.

[0124] After the sample substrate 300 receives the light beam 20a from the light source 400 and emits the light beam 20b toward the filter array 200, the filter array 200 filters the light beam 20b into a light beam 20c having a first wavelength range, a light beam 20c′ having an overlapping wavelength range, and a light beam 20c″ having a second wavelength range. Each photodiode 110 corresponds to one of the multiple filters in the filter array 200, so that the photodiode array 100 can simultaneously provide spectral intensities of three different wavelength ranges through the multiple photodiodes 110.

[0125] Can be used Figures 9A to 9B The biosensor 20 performs Figure 8 Imaging method S200 for detecting spectral shift. Imaging method S200 is similar to Figure 1The imaging method S100 is different from the imaging method S200, but the imaging method S200 can obtain different numbers of image signals based on the adjusted filter array 200. The biosensor 20 in the initial state (for example, step S202) simultaneously generates a first image signal corresponding to the initial light beam 20c, a second image signal corresponding to the initial light beam 20c′, and a third image signal corresponding to the initial light beam 20c″. After the biological sample is placed or reacted in the biosensor 20 (for example, step S204), the biosensor 20 simultaneously generates a fourth image signal corresponding to the late light beam 20c, a fifth image signal corresponding to the late light beam 20c′, and a sixth image signal corresponding to the late light beam 20c″. Next, in step S206, the spectral shift is determined by comparing the differences between the first image signal, the second image signal and the third image signal and the fourth image signal, the fifth image signal and the sixth image signal.

[0126] According to one embodiment of the present disclosure, Figures 10A to 10C Draw the corresponding Figure 8 Imaging methods S200 and Figures 9A to 9B Spectra 360 to 364 of the biosensor 20. Figures 10A to 10C As shown in FIG. 1 , the first filter 210 and the third filter 212 are the same short-bandpass filters, while the second filter 220 and the fourth filter 222 are the same long-bandpass filters partially overlapping the wavelength range of the first filter 210 . Figures 10A to 10C The dotted area in represents the wavelength range of the double-layer filter 230. Figures 10A to 10C The spectral details are similar to Figures 3A to 3C , but obtain Figures 10A to 10C The signal value γ is different from Figures 3A to 3C .

[0127] The first to sixth image signals are used to obtain signal values ​​γ representing the positions of spectrum 360 to spectrum 364. Specifically, the signal value γ can be obtained by the following formula (III), where R is a balance parameter, I S is the image intensity of the image signal filtered by the first filter 210 (ie, the first image signal and the fourth image signal), I A IL is the image intensity of the image signal filtered by the double-layer filter 230 (ie, the second image signal and the fifth image signal), and IL is the image intensity of the image signal filtered by the fourth filter 222 (ie, the third image signal and the sixth image signal).

[0128]

[0129] If RI A =I L -RI A , the signal value γ can be obtained by the following formula (IV) instead.

[0130]

[0131] For spectrum 360, a signal value γ1 equal to 0 can be obtained by formula (III). When spectrum 360 is shifted to spectrum 362, the spectrum red shift causes the signal value γ1 to become a positive signal value γ2. When spectrum 360 is shifted to spectrum 364, the spectrum blue shift causes the signal value γ1 to become a negative signal value γ2.

[0132] In some embodiments, the formula (III) and the formula (IV) can be simplified to more easily obtain the signal value γ. L -RI A =0, formula (III) can be simplified to the following formula (V).

[0133]

[0134] When I S -RI A =0, formula (IV) can be simplified to the following formula (VI).

[0135]

[0136] According to one embodiment of the present disclosure, Figures 11A to 11C Draw the corresponding Figure 8 Imaging methods S200 and Figures 9A to 9B Spectra 370 to 374 of the biosensor 20 . Figures 11A to 11C The steps and biosensors are similar to Figures 10A to 10C ,but Figures 11A to 11C The first filter 210 and the third filter 212 are long bandpass filters, while the second filter 220 and the fourth filter 222 are short bandpass filters partially overlapping the wavelength range of the first filter 210 .

[0137] In order to obtain a positive signal value γ when the spectrum 370 is red-shifted to the spectrum 372, and a negative signal value γ when the spectrum 370 is blue-shifted to the spectrum 374, the signal value γ can be obtained by the following formula (VII), wherein R is a balance parameter, I L is the image intensity of the image signal filtered by the first filter 210, I B is the image intensity of the image signal filtered by the double-layer filter 230, I S is the image intensity of the image signal filtered by the fourth filter 222 .

[0138]

[0139] If RI B =I S -RI B, the signal value γ can be obtained by the following formula (VIII) instead.

[0140]

[0141] For spectrum 370, a signal value γ1 equal to 0 can be obtained by formula (VII). When spectrum 370 is shifted to spectrum 372, the spectrum red shift causes the signal value γ1 to become a positive signal value γ2. When spectrum 370 is shifted to spectrum 374, the spectrum blue shift causes the signal value γ1 to become a negative signal value γ2.

[0142] In some embodiments, formula (VII) and formula (VIII) can be simplified to more easily obtain the signal value γ. S -RI B =0, formula (VII) can be simplified to the following formula (IX).

[0143]

[0144] When I L -RI B =0, formula (VIII) can be simplified to the following formula (X).

[0145]

[0146] Figures 9A to 9D The filter array 200 shown in FIG. 1 includes a first filter 210, a fourth filter 222, and overlapping third filters 212 and second filters 220, thereby providing three different image signals simultaneously. In some other embodiments, the filter array 200 may use a combination of more than two filters to provide three different image signals. For example, Figure 12A According to some embodiments of the present disclosure, execution Figure 8 Schematic diagram of a biosensor 30 in the imaging method S200. Figure 12B Draw Figure 12A A partial three-dimensional view of the biosensor 30.

[0147] The biosensor 30 is similar to Figures 9A to 9Ba biosensor 20, but the filter array 200 of the biosensor 30 includes at least one single-layer first filter 210, at least one first double-layer filter 230 including a third filter 212 and a second filter 220 overlapping the third filter 212, and at least one second double-layer filter 260 including a fifth filter 214 and a fourth filter 250 overlapping the fifth filter 214. The first filter 210, the third filter 212, and the fifth filter 214 have the same wavelength range. After the sample substrate 300 receives the light beam 30a from the light source 400 and emits the light beam 30b toward the filter array 200, the first filter 210 filters the light beam 30b into a light beam 30c within the first wavelength range, the first double-layer filter 230 filters the light beam 30b into a light beam 30c' within the overlapping wavelength range of the first wavelength range and the second wavelength range, and the second double-layer filter 260 filters the light beam 30b into a light beam 30c'' within the overlapping wavelength range of the first wavelength range and the third wavelength range. The third wavelength range is different from the first wavelength range, and the third wavelength range partially overlaps with the first wavelength range. Each photodiode 110 corresponds to one of the multiple filters of the filter array 200, so that the photodiode array 100 can simultaneously provide the spectral intensities of three different wavelength ranges through the multiple photodiodes 110.

[0148] According to an embodiment of the present disclosure, Figures 13A to 14C respectively illustrate corresponding Figure 8 imaging method S200 and Figures 12A to 12B the spectra 380 to 394 of the biosensor 30. As Figures 13A to 13C shown, the first filter 210, the third filter 212, and the fifth filter 214 are the same center band-pass filter, the second filter 220 is a short-pass filter that partially overlaps the wavelength range of the first filter 210, and the fourth filter 250 is a long-pass filter that partially overlaps the wavelength range of the first filter 210. Figures 13A to 13C The dot areas in

[0149] represent the wavelength ranges of the first double-layer filter 230 and the second double-layer filter 260. Figure 13A Referring to Figure 13AAs shown in , for the biosensor 30 in the initial state, the difference between the peak wavelength of the spectrum 380 and the central wavelength of the first filter 210, the difference between the peak wavelength of the spectrum 380 and the cutoff wavelength of the second filter 220, and the difference between the peak wavelength of the spectrum 380 and the starting wavelength of the fourth filter 250 can be within plus or minus 5 nanometers. The slopes of the transition bands of the first filter 210, the second filter 220, and the fourth filter 250 are steeper than the slope of the spectrum 380.

[0150] refer to Figure 13B and Figure 13C After placing or reacting the biological sample, the biosensor 30 simultaneously obtains a fourth image signal corresponding to the late light beam 30c, a fifth image signal corresponding to the late light beam 30c′, and a sixth image signal corresponding to the late light beam 30c″. Then, the spectral shift is determined using the signal value γ obtained by the following formula (XI), where R is a balance parameter, I C is the image intensity of the image signal filtered by the first filter 210 (ie, the first image signal and the fourth image signal), I B is the image intensity of the image signal filtered by the first double-layer filter 230 (ie, the second image signal and the fifth image signal), I A is the image intensity of the image signal filtered by the second double-layer filter 260 (ie, the third image signal and the sixth image signal).

[0151]

[0152] For spectrum 380, a signal value γ1 equal to 0 can be obtained by formula (XI). When spectrum 380 is shifted to spectrum 382, ​​the spectrum red shift causes the signal value γ1 to become a positive signal value γ2. When spectrum 380 is shifted to spectrum 384, the spectrum blue shift causes the signal value γ1 to become a negative signal value γ2.

[0153] refer to Figures 14A to 14C , the first filter 210, the third filter 212 and the fifth filter 214 are the same central bandpass filters, the second filter 220 is a short bandpass filter partially overlapping the wavelength range of the first filter 210, and the fourth filter 250 is a long bandpass filter partially overlapping the wavelength range of the first filter 210. Since the wavelength ranges of the first filter 210, the first double-layer filter 230 and the second double-layer filter 260 and the position of the spectrum 390 are similar Figures 13A to 13C , therefore, the signal value γ of spectrum 390 to spectrum 394 can be obtained by formula (XI).

[0154] According to the above-mentioned embodiment, the imaging method for detecting spectral displacement includes receiving a light beam and simultaneously obtaining a plurality of image signals having different wavelength ranges, and determining the spectral displacement by obtaining a signal value through the image intensity of the image signal. Since a plurality of image signals are obtained simultaneously, the signal-to-noise ratio of the signal value can be increased to improve the detection accuracy, and the immediate output of the signal value can be used for real-time detection of spectral displacement. In addition, the biosensor filters the image signal through a filter array including a single-layer filter and a double-layer filter, thereby miniaturizing the biosensor and increasing the flexibility of the spectral displacement detection range.

[0155] The features of some embodiments are summarized above so that those skilled in the art can better understand the viewpoints of the present disclosure. Those skilled in the art should understand that they can easily use the present disclosure as a basis for designing or modifying other processes and structures to achieve the same purpose and / or achieve the same advantages as the embodiments described herein. Those skilled in the art should also understand that such equivalent constructions do not depart from the spirit and scope of the present disclosure, and various changes, substitutions and modifications can be made without departing from the spirit and scope of the present disclosure.

Claims

1. An imaging method for detecting spectral shift, characterized in that: include: receiving a first light beam through a first photodiode and a second photodiode to simultaneously obtain a first image signal having a first image intensity from the first photodiode and a second image signal having a second image intensity from the second photodiode, The first image signal corresponds to the first light beam filtered by a first filter, the second image signal corresponds to the first light beam filtered by a first double-layer filter, the first double-layer filter includes a second filter overlapped on a third filter, a wavelength range of the third filter is the same as a wavelength range of the first filter, wherein a difference between a peak wavelength of the first image signal and a central wavelength of the first filter is within plus or minus 5 nanometers, wherein a difference between the peak wavelength of the first image signal and a start / cutoff wavelength of the second filter is within plus or minus 5 nanometers; receiving a second light beam through the first photodiode and the second photodiode to simultaneously obtain a third image signal having a third image intensity from the first photodiode and a fourth image signal having a fourth image intensity from the second photodiode, wherein the third image signal corresponds to the second light beam filtered by the first filter, and the fourth image signal corresponds to the second light beam filtered by the first double-layer filter; and The difference between the first image intensity and the second image intensity and the third image intensity and the fourth image intensity is compared to determine a spectral shift from the first light beam to the second light beam.

2. The imaging method according to claim 1, characterized in that When the first light beam is received by the first photodiode and the second photodiode, the second image intensity is half of the first image intensity, a slope of a transition band of the first filter and a slope of a transition band of the second filter are steeper than a slope of the first image signal, and a wavelength range of the first image signal completely falls within the wavelength range of the first filter.

3. The imaging method according to claim 1, characterized in that: The first filter is a short bandpass filter having a bandwidth in the range of 20 nanometers to 60 nanometers, and the second filter is a long wave pass filter or a long bandpass filter.

4. The imaging method according to claim 3, characterized in that: Determining the spectral shift from the first light beam to the second light beam comprises: A signal value γ1 and a signal value γ2 are obtained by formula (I): Where R is the equilibrium parameter, for the signal value γ1, I S and I A is the first image intensity and the second image intensity, and for the signal value γ2, I S and I A are the third image intensity and the fourth image intensity, Wherein when the first light beam is received by the first photodiode and the second photodiode, the signal value γ1 is equal to 0; and The spectral shift is determined by the difference between the signal value γ1 and the signal value γ2.

5. The imaging method according to claim 1, characterized in that: The first filter is a long bandpass filter, and the second filter is a short wave pass filter or a short bandpass filter; and Wherein determining the spectral shift from the first light beam to the second light beam comprises: A signal value γ1 and a signal value γ2 are obtained by formula (II): Where R is the equilibrium parameter, for the signal value γ1, I L and I B is the first image intensity and the second image intensity, and for the signal value γ2, I L and I B are the third image intensity and the fourth image intensity; and The spectral shift is determined by the difference between the signal value γ1 and the signal value γ2.

6. The imaging method according to claim 1, characterized in that: Receiving the first light beam through the first photodiode and the second photodiode further includes receiving the first light beam through a third photodiode to simultaneously obtain a fifth image signal having a fifth image intensity from the third photodiode, the fifth image signal corresponding to the first light beam filtered by a fourth filter, a wavelength range of the fourth filter being the same as a wavelength range of the second filter; and Receiving the second light beam through the first photodiode and the second photodiode further includes receiving the second light beam through the third photodiode to simultaneously obtain a sixth image signal with a sixth image intensity from the third photodiode, and the sixth image signal corresponds to the second light beam filtered by the fourth filter.

7. The imaging method according to claim 6, characterized in that: The first filter is a short bandpass filter, and the second filter is a long bandpass filter; and Wherein determining the spectral shift from the first light beam to the second light beam comprises: A signal value γ1 and a signal value γ2 are obtained by formula (III) or formula (IV): Where R is the balance parameter, for the signal value γ1 (I S , I A , I L ) are the first image intensity, the second image intensity and the fifth image intensity. For the signal value γ2, (I S , I A , I L ) are the third image intensity, the fourth image intensity and the sixth image intensity; and The spectral shift is determined by the difference between the signal value γ1 and the signal value γ2.

8. The imaging method according to claim 7, characterized in that: When I L -RI A =0, the formula (III) is simplified to the formula (V): Among them, when I S -RI A =0, the formula (IV) is simplified to the formula (VI):

9. The imaging method according to claim 6, characterized in that: The first filter is a long bandpass filter, and the second filter is a short bandpass filter; and Wherein determining the spectral shift from the first light beam to the second light beam comprises: A signal value γ1 and a signal value γ2 are obtained by formula (VII) or formula (VIII): Where R is the balance parameter, for the signal value γ1 (I L , I B , I S ) are the first image intensity, the second image intensity and the fifth image intensity. For the signal value γ2, (I L , I B , I S ) are the third image intensity, the fourth image intensity and the sixth image intensity; and The spectral shift is determined by the difference between the signal value γ1 and the signal value γ2.

10. The imaging method according to claim 9, characterized in that: When I S -RI B =0, the formula (VII) is simplified to the formula (IX): Among them, when I L -RI B =0, the formula (VIII) is simplified to the formula (X):

11. The imaging method according to claim 1, wherein: Receiving the first light beam through the first photodiode and the second photodiode further includes receiving the first light beam through a third photodiode to simultaneously obtain a fifth image signal having a fifth image intensity from the third photodiode, the fifth image signal corresponding to the first light beam filtered by a second double-layer filter, the second double-layer filter including a fourth filter superimposed on a fifth filter, a wavelength range of the fifth filter being the same as the wavelength range of the first filter; and Wherein receiving the second light beam through the first photodiode and the second photodiode further includes receiving the second light beam through the third photodiode to simultaneously obtain a sixth image signal having a sixth image intensity from the third photodiode, wherein the sixth image signal corresponds to the second light beam filtered by the second double-layer filter.

12. The imaging method according to claim 11, wherein: The first filter is a central bandpass filter, the second filter is a short-wave pass filter or a short-bandpass filter, and the fourth filter is a long-wave pass filter or a long-bandpass filter; as well as Wherein determining the spectral shift from the first light beam to the second light beam comprises: A signal value γ1 and a signal value γ2 are obtained by formula (XI): Where R is the balance parameter, for the signal value γ1 (I C , I B , I A ) are the first image intensity, the second image intensity and the fifth image intensity. For the signal value γ2, (I C , I B , I A ) are the third image intensity, the fourth image intensity and the sixth image intensity; and The spectral shift is determined by the difference between the signal value γ1 and the signal value γ2.

13. A biosensor for performing the imaging method of claim 1, characterized in that: include: a photodiode array, comprising the first photodiode and the second photodiode; A filter array above the photodiode array includes the first filter and the first double-layer filter, The first photodiode and the second photodiode correspond to the first filter and the first double-layer filter respectively; a sample substrate above the filter array; and A light source is disposed above the sample substrate, wherein the light source is configured to generate a surface plasmon resonance spectrum, and wherein a wavelength range of the light source covers multiple wavelength ranges of the filter array.

Images