US20230119978A1 - Biomolecular image sensor and method thereof for detecting biomolecule - Google Patents

Biomolecular image sensor and method thereof for detecting biomolecule Download PDF

Info

Publication number
US20230119978A1
US20230119978A1 US17/731,840 US202217731840A US2023119978A1 US 20230119978 A1 US20230119978 A1 US 20230119978A1 US 202217731840 A US202217731840 A US 202217731840A US 2023119978 A1 US2023119978 A1 US 2023119978A1
Authority
US
United States
Prior art keywords
image sensor
biomolecule
biomolecular
microstructures
incident light
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
US17/731,840
Inventor
Ping-Hung Yin
Chun-I Shao
Hsiao-Wen Sun
Hsu-Wen Fu
Chang-Hsien Tsai
Lu-Lang Hsu
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Guangzhou Tyrafos Semiconductor Technologies Co Ltd
Original Assignee
Guangzhou Tyrafos Semiconductor Technologies Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Guangzhou Tyrafos Semiconductor Technologies Co Ltd filed Critical Guangzhou Tyrafos Semiconductor Technologies Co Ltd
Priority to US17/731,840 priority Critical patent/US20230119978A1/en
Assigned to Guangzhou Tyrafos Semiconductor Technologies Co., Ltd. reassignment Guangzhou Tyrafos Semiconductor Technologies Co., Ltd. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: FU, HSU-WEN, HSU, LU-LANG, SHAO, CHUN-I, SUN, HSIAO-WEN, TSAI, CHANG-HSIEN, YIN, PING-HUNG
Publication of US20230119978A1 publication Critical patent/US20230119978A1/en
Pending legal-status Critical Current

Links

Images

Classifications

    • H—ELECTRICITY
    • H01—ELECTRIC ELEMENTS
    • H01L—SEMICONDUCTOR DEVICES NOT COVERED BY CLASS H10
    • H01L27/00—Devices consisting of a plurality of semiconductor or other solid-state components formed in or on a common substrate
    • H01L27/14—Devices consisting of a plurality of semiconductor or other solid-state components formed in or on a common substrate including semiconductor components sensitive to infrared radiation, light, electromagnetic radiation of shorter wavelength or corpuscular radiation and specially adapted either for the conversion of the energy of such radiation into electrical energy or for the control of electrical energy by such radiation
    • H01L27/144—Devices controlled by radiation
    • H01L27/146—Imager structures
    • H01L27/14601—Structural or functional details thereof
    • H01L27/14603—Special geometry or disposition of pixel-elements, address-lines or gate-electrodes
    • H01L27/14605—Structural or functional details relating to the position of the pixel elements, e.g. smaller pixel elements in the center of the imager compared to pixel elements at the periphery
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6452—Individual samples arranged in a regular 2D-array, e.g. multiwell plates
    • G01N21/6454—Individual samples arranged in a regular 2D-array, e.g. multiwell plates using an integrated detector array
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/76—Chemiluminescence; Bioluminescence
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54386—Analytical elements
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/0098—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1009—Characterised by arrangements for controlling the aspiration or dispense of liquids
    • G01N35/1011—Control of the position or alignment of the transfer device
    • H—ELECTRICITY
    • H01—ELECTRIC ELEMENTS
    • H01L—SEMICONDUCTOR DEVICES NOT COVERED BY CLASS H10
    • H01L27/00—Devices consisting of a plurality of semiconductor or other solid-state components formed in or on a common substrate
    • H01L27/14—Devices consisting of a plurality of semiconductor or other solid-state components formed in or on a common substrate including semiconductor components sensitive to infrared radiation, light, electromagnetic radiation of shorter wavelength or corpuscular radiation and specially adapted either for the conversion of the energy of such radiation into electrical energy or for the control of electrical energy by such radiation
    • H01L27/144—Devices controlled by radiation
    • H01L27/146—Imager structures
    • H01L27/14601—Structural or functional details thereof
    • H01L27/14625—Optical elements or arrangements associated with the device
    • H01L27/14627—Microlenses
    • H—ELECTRICITY
    • H01—ELECTRIC ELEMENTS
    • H01L—SEMICONDUCTOR DEVICES NOT COVERED BY CLASS H10
    • H01L27/00—Devices consisting of a plurality of semiconductor or other solid-state components formed in or on a common substrate
    • H01L27/14—Devices consisting of a plurality of semiconductor or other solid-state components formed in or on a common substrate including semiconductor components sensitive to infrared radiation, light, electromagnetic radiation of shorter wavelength or corpuscular radiation and specially adapted either for the conversion of the energy of such radiation into electrical energy or for the control of electrical energy by such radiation
    • H01L27/144—Devices controlled by radiation
    • H01L27/146—Imager structures
    • H01L27/14601—Structural or functional details thereof
    • H01L27/1464—Back illuminated imager structures
    • H—ELECTRICITY
    • H01—ELECTRIC ELEMENTS
    • H01L—SEMICONDUCTOR DEVICES NOT COVERED BY CLASS H10
    • H01L27/00—Devices consisting of a plurality of semiconductor or other solid-state components formed in or on a common substrate
    • H01L27/14—Devices consisting of a plurality of semiconductor or other solid-state components formed in or on a common substrate including semiconductor components sensitive to infrared radiation, light, electromagnetic radiation of shorter wavelength or corpuscular radiation and specially adapted either for the conversion of the energy of such radiation into electrical energy or for the control of electrical energy by such radiation
    • H01L27/144—Devices controlled by radiation
    • H01L27/146—Imager structures
    • H01L27/14643—Photodiode arrays; MOS imagers
    • H01L27/14645—Colour imagers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/025—Align devices or objects to ensure defined positions relative to each other
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027—General features of the devices
    • G01N2035/103—General features of the devices using disposable tips
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2201/00—Features of devices classified in G01N21/00
    • G01N2201/04—Batch operation; multisample devices
    • G01N2201/0407—Batch operation; multisample devices with multiple optical units, e.g. one per sample

Definitions

  • the present invention relates to a biomolecular image sensor and method thereof for detecting biomolecule, and more particularly to a biomolecular image sensor with a plurality of microstructures repeatedly arranged on a surface of an image sensing element, and method thereof for detecting biomolecule.
  • Enzyme-linked immunosorbent assay or enzyme-linked immunoassay (EIA) is the specific antigen-antibody reaction tests.
  • the specific binding properties between antigens and antibodies are used to detect the molecules in samples.
  • the presence of specific antigens or antibodies may be shown by the color reaction with enzymes. Quantitative analysis may be carried out by the depth of color to achieve the detection and screening.
  • biochemical tests such as on tissue sections require large and expensive equipment to receive optical or electronic signals for analyzing the status of biochemical molecular reactions, for example, observing with microscopes and capturing images with additional photo equipment for further analysis, which require time and manual operations.
  • biochips have to be additionally equipped with other expensive and large image capture systems or equipment to detect and capture the luminescent images of the biochips after the biochemical detection process for subsequent analysis.
  • traditional ELISA has to be equipped with an ELISA reader to detect the absorbance in each well of the micro-well plate after the operation with ELISA kit for quantification.
  • POCT point of care testing
  • One objective of the present invention is to provide a biomolecular image sensor, comprising: an image sensing element, containing a plurality of unit pixels disposed in an array on a substrate, wherein each of the plurality of unit pixels contains at least one photoelectric conversion element, the photoelectric conversion element receives an incident light to generate electrons, and a surface of the image sensing element receiving the incident light is defined as a light receiving surface; and a microstructure layer, disposed on the light receiving surface of the image sensing element and having a plurality of microstructures arranged in a specific shape repeatedly, wherein each of the plurality of microstructures corresponds to at least one unit pixel.
  • each of the microstructures is formed on the image sensor element by photolithography process or imprint lithography process.
  • the further objective of the present invention is to provide a biomolecular image sensor, comprising: an image sensing element, containing a plurality of unit pixels disposed in an array on a substrate, wherein each of the plurality of unit pixels contains at least one photoelectric conversion element, the photoelectric conversion element receives an incident light to generate electrons, and a surface of the image sensing element receiving the incident light is defined as a light receiving surface; and a microstructure layer, disposed on the light receiving surface of the image sensing element and being an array formed by a plurality of microlenses, wherein a plurality of microstructures are formed between the plurality of microlenses, and each of the plurality of microstructures corresponds to at least one unit pixel.
  • the biomolecular image sensor further comprises at least one readout circuit coupled to the unit pixels, and the readout circuit generates a voltage signal based on the number of the electrons.
  • each of the microlenses is provided to accommodate at least one biomolecule.
  • the incident light is a light emitted by a fluorescent marker or a chemiluminescent marker on the biomolecule.
  • the biomolecular image sensor further comprises a carrier carrying the biomolecule, and the carrier is accommodated in the microstructure, and the carrier may be a microparticle. Further, one microstructure accommodates one carrier chemically bonded with multiple biomolecules.
  • the bottom portions and the side portions of the microstructures are composed of different materials, and each of the microlenses is formed on the image sensor element by photolithography process or imprint lithography process.
  • the other objective of the present invention is to provide a method of detecting a biomolecule, comprising: (a) providing the biomolecular image sensor; (b) accommodating the biomolecule in a sample in each of the plurality of microstructures; (c) detecting an incident light in each of the plurality of microstructures by each of the plurality of unit pixels; (d) generating electrons from the incident light detected by each of the plurality of unit pixels by the photoelectric conversion element; (e) generating a voltage signal based on a number of the electrons by a readout circuit coupled to each of the plurality of unit pixels; and (f) analyzing a presence and/or a concentration of the biomolecule based on the voltage signal; wherein, a fluorescent marker or a chemiluminescent marker is added on the biomolecule before or after (b), and the incident light comprises a light emitted by the fluorescent marker or the chemiluminescent marker.
  • the present invention is a completely innovative image sensor for detecting biomolecules prepared by a semiconductor process.
  • the image sensor of the present invention which is smaller than a coin, may directly detect the presence of specific biomolecules in samples and quantify concentration thereof, but does not require additional large equipment.
  • the present invention does not require additional image capture system or equipment. That is, the biomolecular image sensor of the present invention provides the functions of biological or chemical analysis and image capture and interpretation. That is, the detection process may be directly operated on the biomolecule image sensor of the present invention, and corresponding detection results may be obtained in real time.
  • the biomolecule image sensor of the present invention independently detects the incident light in the corresponding microstructure through each unit pixel, and compares each measured signal readout with the thresholds independently, and therefore, even when the concentration of the biomolecules in the sample is extremely low, the presence and intensity of chemiluminescent or fluorescent signals of the biomolecules would still be accurately interpreted, so as to increase the detection sensitivity.
  • the microstructure in the biomolecule image sensor of the present invention enables the biomolecules to be dispersed evenly on the light receiving surface, the photoelectric conversion element would easily determine the incident light really from the luminescent or fluorescent markers on the biomolecules.
  • the biomolecule image sensor of the present invention provides the quantitative mode of analog colorimetric method and digital method, and could be switched according to different concentrations of biomolecules, so as to maximize the detection range.
  • FIG. 1 shows a cross-section view of the biomolecular image sensor according to one embodiment of the present invention.
  • FIG. 2 shows a schematic view of inverted pyramidal microstructures of the biomolecular image sensor according to another embodiment of the present invention.
  • FIG. 3 shows a schematic view of honeycomb microstructures of the biomolecular image sensor according to another embodiment of the present invention.
  • FIG. 4 A shows a schematic view of the biomolecular image sensor according to another embodiment of the present invention, wherein the microstructure layer is an array formed by a plurality of microlenses and one photoelectric conversion element corresponds to four microlenses.
  • FIG. 4 B shows a cross-sectional view of the biomolecular image sensor shown in FIG. 4 A .
  • FIG. 5 shows a schematic view of the biomolecular image sensor according to another embodiment of the present invention, wherein the microstructure layer is an array formed by a plurality of microlenses and one photoelectric conversion element corresponds to the microstructure formed by four microlenses.
  • FIG. 6 shows a schematic view of the biomolecular image sensor according to another embodiment of the present invention, wherein the microstructure layer is an array formed by a plurality of microlenses and one photoelectric conversion element corresponds to one microlens.
  • FIG. 7 A shows a flowchart of the method of detecting biomolecules through the biomolecular image sensor according to one embodiment of the present invention.
  • FIG. 7 B shows a flowchart of the method of detecting biomolecules through the biomolecular image sensor according to another embodiment of the present invention.
  • the operating procedures and parameter conditions for enzyme-linked immunosorbent assay are within the professional literacy and routine techniques of one with ordinary skill in the art.
  • the fluorescent molecules may be, but not limited to, FITC, HEX, FAM, TAMRA, Cy3, Cy5, quantum dot, or the like, and may be used with a quencher dye.
  • FIG. 1 shows a biomolecular image sensor 100 according to one embodiment of the present invention.
  • the biomolecule image sensor 100 of the present invention comprises: an image sensing element 10 and a microstructure layer 20 , and may further comprise at least one readout circuit; wherein, the image sensing element 10 may contain a plurality of unit pixels 11 disposed in an array on a substrate; further, the microstructure layer 20 may have a plurality of microstructures 21 repeatedly arranged in a specific shape, and may accommodate a biomolecule A, or a carrier 22 carrying with a biomolecule A.
  • the image sensing element 10 in the biomolecular image sensor 100 may be a back-illuminated complementary metal-oxide-semiconductor (CMOS) image sensor or a front-illuminated CMOS image sensor; however, the present invention is not limited thereto.
  • CMOS complementary metal-oxide-semiconductor
  • each of the unit pixels 11 may include at least one photoelectric conversion element, wherein the photoelectric conversion element may generate electrons after receiving an incident light, and the photoelectric conversion element also contains the ability to accumulate the above-mentioned electrons; however, the present invention is not limited to thereto.
  • the photoelectric conversion element may be an element that generates and accumulates electrons corresponding to the incident light.
  • the photoelectric conversion element may be a photodiode, a photo transistor, a photo gate, a pinned photo diode (PPD), an avalanche photodiode (APD), a single-photon avalanche diode (SPAD), a photomultiplier tube (PMT), or any combination thereof.
  • a surface of the image sensing element 10 receiving the incident light may be defined as a light receiving surface, and the microstructure layer 20 may be disposed on the light receiving surface.
  • each of the microstructures 21 in the microstructure layer 20 may correspond to the unit pixel 11 respectively. In this way, since the microstructures 21 have a corresponding relationship with the unit pixels 11 , the photoelectric conversion elements in the unit pixels 11 may receive the incident light corresponding to the microstructures 21 .
  • the biomolecular image sensor 100 of the present invention may be formed by a semiconductor process. More specifically, the microstructures 21 may be formed on the image sensing element 10 by photolithography process or imprint lithography process, and not just general by surface treatment or coatings.
  • the readout circuits may be coupled to the unit pixels 11 and generate a voltage signal, which is used as the signal readout, according to the number of the electrons generated after the photoelectric conversion element receives the incident light.
  • the biomolecular image sensor 100 of the present invention is not limited to any specific applications.
  • the biomolecule image sensor 100 of the present invention is used for biological or chemical analysis, such as detecting the presence and/or concentration of a biomolecule A in a sample. That is, the incident light may be a light emitted by a marker, a fluorescent marker, a reporter marker, or a chemiluminescent marker of the biomolecule A. In other words, the presence and concentration of the biomolecule A may be analyzed by detecting the markers.
  • the biomolecule image sensor 100 of the present invention when used to detect the biomolecule A, in the biological or chemical analysis process, the biomolecule A may undergo luminescence reaction with other molecules and emit an incident light such as chemiluminescence or fluorescence.
  • the biomolecule A may be a protein, a peptide, an antibody, a nucleic acid, or the like.
  • the microstructures 21 may be disposed to accommodate the biomolecules A, and the biomolecules A in the sample may be dispersed in a plurality of the microstructures 21 . Since the single microstructure 21 corresponds to the single unit pixel 11 , the photoelectric conversion element in the single unit pixel 11 would only receive the light emitted by the biomolecules A in the single microstructure 21 . Therefore, the sensitivity of detecting the specific biomolecule A in the sample would be improved.
  • the biomolecule image sensor 100 of the present invention may be used with a carrier 22 for carrying the biomolecule A, so that the biomolecule A may be more easily accommodated in the microstructures 21 , and the luminescence reaction of the biomolecule A may be easily performed.
  • the carrier 22 may be a microparticle, and the microparticle may be used for performing ELISA.
  • the biomolecule A is linked on the microparticle for performing the luminescence reaction to generate a chemiluminescence, and the chemiluminescence is the incident light received by the photoelectric conversion element in the unit pixel 11 .
  • the biomolecule A such as an antibody may be linked to the microparticle by forming an amide bond through the EDC/NHS reaction.
  • the microparticle is preferably a magnetic bead with 1 to 3 ⁇ m diameter.
  • the magnetic bead may be prepared by magnetic materials of magnetic elements such as Fe, Ni, Co, ferromagnetic alloys such as Nb-Fe-B, or iron oxides such as Fe 3 O 4 , Fe 2 O 3 , FeO.
  • the microparticle may also be prepared by non-magnetic materials such as Au, sepharose, polystyrene, and SiO 2 .
  • the single microstructure 21 has to accommodate only one carrier 22 as much as possible, so the width/diameter and height/depth of the microstructures 21 have to match the carrier 22 .
  • the diameter of the microstructure is preferably 1.3 to 1.8 times larger than the diameter of the microparticle
  • the depth is preferably 1.2 to 1.3 times larger than the diameter of the microparticle
  • the aspect ratio is preferably 1 to 1.2 times.
  • the microstructures 21 may be grooves with 2.5 to 3 ⁇ m diameter and 2.5 ⁇ m depth, and the spacing between each microstructure is 2-3 ⁇ m.
  • surfactant may be used to improve the degree of dispersion thereof.
  • the biomolecule A may be detected by an antibody or aptamer labeled with a fluorescent molecule.
  • a radiated light is generated by irradiating excitation light with a specific wavelength, and the radiated light is the incident light received by the photoelectric conversion element in the unit pixel 11 .
  • Antibodies or aptamers labeled with different fluorescent molecules may be contained on the same carrier 22 , so that the detection of various target biomolecules may be performed on the same sample during one single detection.
  • a CMOS image sensor with RGB technology may be used for antibodies or aptamers containing multiple fluorescent labels.
  • the bottom portions and the side portions of the microstructures 21 may be composed of the same or different materials.
  • a surface of the microstructures 21 in contact with the light receiving surface i.e., the bottom portions
  • a surface of the microstructures 21 not in contact with the light receiving surface i.e., the side portions
  • the bottom portions and the side portions of the microstructures 21 may both be composed of SiO 2 .
  • microstructures 21 may be implemented in different embodiments to allow the carriers 22 to be more easily accommodated in the microstructures 21 , taking into account the influence of air pressure or hydrodynamics on the carriers 22 during operation.
  • FIG. 2 shows a biomolecule image sensor 100 A according to another embodiment of the present invention.
  • the microstructure layer 20 A may be an array formed by a plurality of inverted pyramidal microstructures 21 A, which facilitates the gathering of the incident light, and the inverted pyramidal microstructures 21 A are preferably with 2.5 ⁇ m length and width.
  • FIG. 3 shows a biomolecule image sensor 100 B according to another embodiment of the present invention.
  • the microstructure layer 20 B may be an array formed by a plurality of honeycomb microstructures 21 B. The most densely distributed without gaps between the honeycomb microstructures 21 B is favorable for the carrier 22 to be evenly accommodated therein.
  • the honeycomb microstructures 21 B are preferably with 1.55 ⁇ m length and 2.5 ⁇ m depth.
  • FIGS. 4 A and 4 B show a biomolecule image sensor 200 according to another embodiment of the present invention.
  • the microstructure layer 40 may be an array formed by a plurality of microlenses 42 , wherein a plurality of microstructures 41 are formed between the microlenses 42 to accommodate the biomolecule A or the carrier 22 carrying the biomolecule A.
  • the microlenses 42 and the unit pixels 11 may be correspondingly arranged in different ways.
  • each microlenses 42 may be arranged corresponding to one single unit pixel 11 , wherein the area of the unit pixel 11 may be greater than or equal to the area surrounded by the four microlenses 42 .
  • the area of the unit pixel 11 A may be smaller than the area surrounded by the four microlenses 42 , and the microstructure 41 formed by the four microlenses 42 is arranged corresponding to the unit pixel 11 A.
  • each microlens 42 may be arranged corresponding to one single unit pixel 11 B respectively.
  • the method of detecting a biomolecule in a sample by the biomolecule image sensor of the present invention may be as following.
  • a fluorescent marker or a chemiluminescent marker is added to the biomolecules in the sample through biological or chemical analysis such as ELISA, and the biomolecules are accommodated in the microstructures (S 11 ).
  • the incident light in one single microstructure is detected by the unit pixels respectively (S 12 ), wherein the incident light includes the lights emitted by the fluorescent maker or the chemiluminescent marker of the biomolecule.
  • Electrons are generated from the incident light received by each of the unit pixels through the photoelectric conversion element (S 13 ).
  • a voltage signal is generated according to the number of the electrons through the readout circuits (S 14 ). The presence and/or concentration of the biomolecule are then analyzed according to the voltage signal.
  • the biomolecule image sensor of the present invention when used to detect a biomolecule in a sample, the biomolecule containing fluorescent makers or chemiluminescent markers may also be placed on a substrate such as a glass slide, traditional biochips, or the like.
  • the substrate is then placed on the biomolecule image sensor of the present invention, so that the unit pixels would detect an incident light from the corresponding position respectively.
  • electrons are also generated from the incident light through the photoelectric conversion element, and a voltage signal, which is used as the signal readout, is generated according to the number of the electrons through the readout circuits.
  • the presence and/or concentration of the biomolecule are then analyzed according to the voltage signal.
  • the quantification thereof may be not only performed by analog colorimetric method, that is, the incident light in one single microstructure received by the unit pixels is one single readout to determine the presence of the biomolecule (S 151 ) or further compare with the standard curve for obtaining the concentration of the biomolecule (S 161 ), but also by digital method, that is, according to the predetermined threshold (S 152 ), the unit pixels with the signal readout exceeding the threshold are defined as 1 (S 153 ) and the unit pixels with the signal readout not exceeding the threshold are defined as 0 (S 154 ), and the number of unit pixels defined as 1 is calculated and compared with the standard curve (S 162 ) to obtain the concentration of the biomolecule more accurately.
  • analog colorimetric method that is, the incident light in one single microstructure received by the unit pixels is one single readout to determine the presence of the biomolecule (S 151 ) or further compare with the standard curve for obtaining the concentration of the biomolecule (S 161 )
  • digital method that is, according to the predetermined threshold (S
  • the present invention is a completely innovative image sensor for detecting biomolecules prepared by a semiconductor process.
  • the image sensor of the present invention which is smaller than a coin, may directly detect the presence of specific biomolecules in samples and quantify concentration thereof, but does not require additional large equipment.
  • the present invention does not require additional image capture system or equipment. That is, the biomolecular image sensor of the present invention provides the functions of biological or chemical analysis and image capture and interpretation. That is, the detection process may be directly operated on the biomolecule image sensor of the present invention, and corresponding detection results may be obtained in real time.
  • the biomolecule image sensor of the present invention independently detects the incident light in the corresponding microstructure through each unit pixel, and compares each measured signal readout with the thresholds independently, and therefore, even when the concentration of the biomolecules in the sample is extremely low, the presence and intensity of chemiluminescent or fluorescent signals of the biomolecules would still be accurately interpreted, so as to increase the detection sensitivity.
  • the microstructure in the biomolecule image sensor of the present invention enables the biomolecules to be dispersed evenly on the light receiving surface, the photoelectric conversion element would easily determine the incident light really from the luminescent or fluorescent markers on the biomolecules.
  • the biomolecule image sensor of the present invention provides the quantitative mode of analog colorimetric method and digital method, and could be switched according to different concentrations of biomolecules, so as to maximize the detection range.

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Power Engineering (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Pathology (AREA)
  • Condensed Matter Physics & Semiconductors (AREA)
  • Microelectronics & Electronic Packaging (AREA)
  • Computer Hardware Design (AREA)
  • Electromagnetism (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Molecular Biology (AREA)
  • Plasma & Fusion (AREA)
  • Optics & Photonics (AREA)
  • Clinical Laboratory Science (AREA)
  • Biotechnology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Dispersion Chemistry (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)
  • Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)
  • Photometry And Measurement Of Optical Pulse Characteristics (AREA)

Abstract

The invention provides a biomolecular image sensor with a plurality of microstructures repeatedly arranged on a surface of an image sensing element, and method thereof for detecting biomolecule.

Description

    CROSS-REFERENCE TO RELATED APPLICATION
  • This application claims priority of U.S. provisional application No. 63/255,446, filed on Oct. 14, 2021, the content of which is incorporated herein in its entirety by reference.
    • BACKGROUND OF THE INVENTION 1. Field of the Invention
  • The present invention relates to a biomolecular image sensor and method thereof for detecting biomolecule, and more particularly to a biomolecular image sensor with a plurality of microstructures repeatedly arranged on a surface of an image sensing element, and method thereof for detecting biomolecule.
    • 2. The Prior Art
  • Enzyme-linked immunosorbent assay (ELISA) or enzyme-linked immunoassay (EIA) is the specific antigen-antibody reaction tests. The specific binding properties between antigens and antibodies are used to detect the molecules in samples. The presence of specific antigens or antibodies may be shown by the color reaction with enzymes. Quantitative analysis may be carried out by the depth of color to achieve the detection and screening.
  • Biochips are micro devices that use biological materials on a substrate to produce specific biochemical reactions with the biomolecules, and may be quantified by a highly sensitive detection system. Biochips provide fast, accurate, and low cost bioanalytical testing capabilities. Biochips are basically miniaturized substrates that may perform hundreds or thousands of biochemical reactions simultaneously.
  • However, traditional biochemical tests such as on tissue sections require large and expensive equipment to receive optical or electronic signals for analyzing the status of biochemical molecular reactions, for example, observing with microscopes and capturing images with additional photo equipment for further analysis, which require time and manual operations. On the other hand, traditional biochips have to be additionally equipped with other expensive and large image capture systems or equipment to detect and capture the luminescent images of the biochips after the biochemical detection process for subsequent analysis. Further, traditional ELISA has to be equipped with an ELISA reader to detect the absorbance in each well of the micro-well plate after the operation with ELISA kit for quantification.
  • With the increasing popularity of the point of care testing (POCT), a personalized health test with a short analysis time and simple operation, it is necessary to develop a more sensitive and simpler detection device and method to overcome the problems from large equipment and complex biochemical detection processes in traditional biomolecule detection methods.
    • SUMMARY OF THE INVENTION
  • One objective of the present invention is to provide a biomolecular image sensor, comprising: an image sensing element, containing a plurality of unit pixels disposed in an array on a substrate, wherein each of the plurality of unit pixels contains at least one photoelectric conversion element, the photoelectric conversion element receives an incident light to generate electrons, and a surface of the image sensing element receiving the incident light is defined as a light receiving surface; and a microstructure layer, disposed on the light receiving surface of the image sensing element and having a plurality of microstructures arranged in a specific shape repeatedly, wherein each of the plurality of microstructures corresponds to at least one unit pixel.
  • In the preferred embodiment of the present invention, the microstructure layer is an array formed by a plurality of inverted pyramidal or honeycomb microstructures.
  • In the preferred embodiment of the present invention, each of the microstructures is formed on the image sensor element by photolithography process or imprint lithography process.
  • The further objective of the present invention is to provide a biomolecular image sensor, comprising: an image sensing element, containing a plurality of unit pixels disposed in an array on a substrate, wherein each of the plurality of unit pixels contains at least one photoelectric conversion element, the photoelectric conversion element receives an incident light to generate electrons, and a surface of the image sensing element receiving the incident light is defined as a light receiving surface; and a microstructure layer, disposed on the light receiving surface of the image sensing element and being an array formed by a plurality of microlenses, wherein a plurality of microstructures are formed between the plurality of microlenses, and each of the plurality of microstructures corresponds to at least one unit pixel.
  • In the preferred embodiment of the present invention, the biomolecular image sensor further comprises at least one readout circuit coupled to the unit pixels, and the readout circuit generates a voltage signal based on the number of the electrons.
  • In the preferred embodiment of the present invention, each of the microlenses is provided to accommodate at least one biomolecule.
  • In the preferred embodiment of the present invention, the incident light is a light emitted by a fluorescent marker or a chemiluminescent marker on the biomolecule.
  • In the preferred embodiment of the present invention, the biomolecular image sensor further comprises a carrier carrying the biomolecule, and the carrier is accommodated in the microstructure, and the carrier may be a microparticle. Further, one microstructure accommodates one carrier chemically bonded with multiple biomolecules.
  • In the preferred embodiment of the present invention, the bottom portions and the side portions of the microstructures are composed of different materials, and each of the microlenses is formed on the image sensor element by photolithography process or imprint lithography process.
  • The other objective of the present invention is to provide a method of detecting a biomolecule, comprising: (a) providing the biomolecular image sensor; (b) accommodating the biomolecule in a sample in each of the plurality of microstructures; (c) detecting an incident light in each of the plurality of microstructures by each of the plurality of unit pixels; (d) generating electrons from the incident light detected by each of the plurality of unit pixels by the photoelectric conversion element; (e) generating a voltage signal based on a number of the electrons by a readout circuit coupled to each of the plurality of unit pixels; and (f) analyzing a presence and/or a concentration of the biomolecule based on the voltage signal; wherein, a fluorescent marker or a chemiluminescent marker is added on the biomolecule before or after (b), and the incident light comprises a light emitted by the fluorescent marker or the chemiluminescent marker.
  • The present invention is a completely innovative image sensor for detecting biomolecules prepared by a semiconductor process. The image sensor of the present invention, which is smaller than a coin, may directly detect the presence of specific biomolecules in samples and quantify concentration thereof, but does not require additional large equipment.
  • On the other hand, compared with the traditional biochip, the present invention does not require additional image capture system or equipment. That is, the biomolecular image sensor of the present invention provides the functions of biological or chemical analysis and image capture and interpretation. That is, the detection process may be directly operated on the biomolecule image sensor of the present invention, and corresponding detection results may be obtained in real time.
  • On the other hand, compared with the traditional methods of quantifying biomolecules, the biomolecule image sensor of the present invention independently detects the incident light in the corresponding microstructure through each unit pixel, and compares each measured signal readout with the thresholds independently, and therefore, even when the concentration of the biomolecules in the sample is extremely low, the presence and intensity of chemiluminescent or fluorescent signals of the biomolecules would still be accurately interpreted, so as to increase the detection sensitivity.
  • Further, because the microstructure in the biomolecule image sensor of the present invention enables the biomolecules to be dispersed evenly on the light receiving surface, the photoelectric conversion element would easily determine the incident light really from the luminescent or fluorescent markers on the biomolecules. The biomolecule image sensor of the present invention provides the quantitative mode of analog colorimetric method and digital method, and could be switched according to different concentrations of biomolecules, so as to maximize the detection range.
  • In order to enable one with ordinary skill in the art to understand the purpose, features, and functions of the present invention, the invention is described in detail below by means of the following specific embodiments and with the attached drawings.
    • BRIEF DESCRIPTION OF THE DRAWINGS
  • FIG. 1 shows a cross-section view of the biomolecular image sensor according to one embodiment of the present invention.
  • FIG. 2 shows a schematic view of inverted pyramidal microstructures of the biomolecular image sensor according to another embodiment of the present invention.
  • FIG. 3 shows a schematic view of honeycomb microstructures of the biomolecular image sensor according to another embodiment of the present invention.
  • FIG. 4A shows a schematic view of the biomolecular image sensor according to another embodiment of the present invention, wherein the microstructure layer is an array formed by a plurality of microlenses and one photoelectric conversion element corresponds to four microlenses.
  • FIG. 4B shows a cross-sectional view of the biomolecular image sensor shown in FIG. 4A.
  • FIG. 5 shows a schematic view of the biomolecular image sensor according to another embodiment of the present invention, wherein the microstructure layer is an array formed by a plurality of microlenses and one photoelectric conversion element corresponds to the microstructure formed by four microlenses.
  • FIG. 6 shows a schematic view of the biomolecular image sensor according to another embodiment of the present invention, wherein the microstructure layer is an array formed by a plurality of microlenses and one photoelectric conversion element corresponds to one microlens.
  • FIG. 7A shows a flowchart of the method of detecting biomolecules through the biomolecular image sensor according to one embodiment of the present invention.
  • FIG. 7B shows a flowchart of the method of detecting biomolecules through the biomolecular image sensor according to another embodiment of the present invention.
    •  DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
  • The embodiments of the present invention are further described with the following drawings. The following embodiments are given to illustrate the present invention and are not intended to limit the scope of the present invention, and one with ordinary skill in the art may make some modifications and refinements without departing from the spirit and scope of the present invention. Therefore, the scope of the present invention is defined by the scope of the appended claims.
  • The terms used herein are only for describing the embodiments, and are not intended to limit the present invention. Unless otherwise defined, the terms have meanings commonly understood by one with ordinary skill in the art to which the present invention belongs. As used herein, the singular forms “a” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. As used herein, the term “and/or” includes any and all combinations of one or more of the associated listed items.
  • Similarly, when an element is referred to be “on” another element, it would be understood that the element may be directly on the other element or intermediate elements may be present. In contrast, the term “directly” represents that there is no intermediate element. The term “comprising” used herein should be understood to indicate the presence of stated features, integers, steps, operations, elements, and/or components, but not exclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or any combination thereof.
  • According to the present invention, the operating procedures and parameter conditions for enzyme-linked immunosorbent assay (ELISA) are within the professional literacy and routine techniques of one with ordinary skill in the art.
  • According to the present invention, the fluorescent molecules may be, but not limited to, FITC, HEX, FAM, TAMRA, Cy3, Cy5, quantum dot, or the like, and may be used with a quencher dye.
  • FIG. 1 shows a biomolecular image sensor 100 according to one embodiment of the present invention. As shown in FIG. 1 , the biomolecule image sensor 100 of the present invention comprises: an image sensing element 10 and a microstructure layer 20, and may further comprise at least one readout circuit; wherein, the image sensing element 10 may contain a plurality of unit pixels 11 disposed in an array on a substrate; further, the microstructure layer 20 may have a plurality of microstructures 21 repeatedly arranged in a specific shape, and may accommodate a biomolecule A, or a carrier 22 carrying with a biomolecule A.
  • More specifically, the image sensing element 10 in the biomolecular image sensor 100 according to the present invention may be a back-illuminated complementary metal-oxide-semiconductor (CMOS) image sensor or a front-illuminated CMOS image sensor; however, the present invention is not limited thereto.
  • In the embodiments of the present invention, each of the unit pixels 11 may include at least one photoelectric conversion element, wherein the photoelectric conversion element may generate electrons after receiving an incident light, and the photoelectric conversion element also contains the ability to accumulate the above-mentioned electrons; however, the present invention is not limited to thereto.
  • Further, the photoelectric conversion element may be an element that generates and accumulates electrons corresponding to the incident light. For example, the photoelectric conversion element may be a photodiode, a photo transistor, a photo gate, a pinned photo diode (PPD), an avalanche photodiode (APD), a single-photon avalanche diode (SPAD), a photomultiplier tube (PMT), or any combination thereof.
  • In the embodiments of the present invention, a surface of the image sensing element 10 receiving the incident light may be defined as a light receiving surface, and the microstructure layer 20 may be disposed on the light receiving surface. In the preferred embodiments of the present invention, each of the microstructures 21 in the microstructure layer 20 may correspond to the unit pixel 11 respectively. In this way, since the microstructures 21 have a corresponding relationship with the unit pixels 11, the photoelectric conversion elements in the unit pixels 11 may receive the incident light corresponding to the microstructures 21.
  • The biomolecular image sensor 100 of the present invention may be formed by a semiconductor process. More specifically, the microstructures 21 may be formed on the image sensing element 10 by photolithography process or imprint lithography process, and not just general by surface treatment or coatings.
  • In the embodiments of the present invention, the readout circuits may be coupled to the unit pixels 11 and generate a voltage signal, which is used as the signal readout, according to the number of the electrons generated after the photoelectric conversion element receives the incident light.
  • The biomolecular image sensor 100 of the present invention is not limited to any specific applications. In one preferred embodiment, the biomolecule image sensor 100 of the present invention is used for biological or chemical analysis, such as detecting the presence and/or concentration of a biomolecule A in a sample. That is, the incident light may be a light emitted by a marker, a fluorescent marker, a reporter marker, or a chemiluminescent marker of the biomolecule A. In other words, the presence and concentration of the biomolecule A may be analyzed by detecting the markers. More specifically, when the biomolecule image sensor 100 of the present invention is used to detect the biomolecule A, in the biological or chemical analysis process, the biomolecule A may undergo luminescence reaction with other molecules and emit an incident light such as chemiluminescence or fluorescence. Further, the biomolecule A may be a protein, a peptide, an antibody, a nucleic acid, or the like.
  • More specifically, the microstructures 21 may be disposed to accommodate the biomolecules A, and the biomolecules A in the sample may be dispersed in a plurality of the microstructures 21. Since the single microstructure 21 corresponds to the single unit pixel 11, the photoelectric conversion element in the single unit pixel 11 would only receive the light emitted by the biomolecules A in the single microstructure 21. Therefore, the sensitivity of detecting the specific biomolecule A in the sample would be improved.
  • Further, the biomolecule image sensor 100 of the present invention may be used with a carrier 22 for carrying the biomolecule A, so that the biomolecule A may be more easily accommodated in the microstructures 21, and the luminescence reaction of the biomolecule A may be easily performed. For example, the carrier 22 may be a microparticle, and the microparticle may be used for performing ELISA. The biomolecule A is linked on the microparticle for performing the luminescence reaction to generate a chemiluminescence, and the chemiluminescence is the incident light received by the photoelectric conversion element in the unit pixel 11. The biomolecule A such as an antibody may be linked to the microparticle by forming an amide bond through the EDC/NHS reaction.
  • More specifically, the microparticle is preferably a magnetic bead with 1 to 3 µm diameter. The magnetic bead may be prepared by magnetic materials of magnetic elements such as Fe, Ni, Co, ferromagnetic alloys such as Nb-Fe-B, or iron oxides such as Fe3O4, Fe2O3, FeO. Alternatively, the microparticle may also be prepared by non-magnetic materials such as Au, sepharose, polystyrene, and SiO2.
  • More specifically, the single microstructure 21 has to accommodate only one carrier 22 as much as possible, so the width/diameter and height/depth of the microstructures 21 have to match the carrier 22. More specifically, the diameter of the microstructure is preferably 1.3 to 1.8 times larger than the diameter of the microparticle, the depth is preferably 1.2 to 1.3 times larger than the diameter of the microparticle, and the aspect ratio is preferably 1 to 1.2 times. For example, when the carrier 22 is a microparticle with 2 µm diameter, the microstructures 21 may be grooves with 2.5 to 3 µm diameter and 2.5 µm depth, and the spacing between each microstructure is 2-3 µm. To avoid the molecular forces between the carriers 22 to cause them stack on each other and fail to disperse into a single one, which further causes the single carrier 22 difficult to accommodate in one single microstructure 21, surfactant, external magnetic field, or vibration to destroy the force between the carriers 22 may be used to improve the degree of dispersion thereof.
  • Alternatively, the biomolecule A may be detected by an antibody or aptamer labeled with a fluorescent molecule. A radiated light is generated by irradiating excitation light with a specific wavelength, and the radiated light is the incident light received by the photoelectric conversion element in the unit pixel 11. Antibodies or aptamers labeled with different fluorescent molecules may be contained on the same carrier 22, so that the detection of various target biomolecules may be performed on the same sample during one single detection. Further, a CMOS image sensor with RGB technology may be used for antibodies or aptamers containing multiple fluorescent labels.
  • In the biomolecular image sensor 100 of the present invention, the bottom portions and the side portions of the microstructures 21 may be composed of the same or different materials. For example, a surface of the microstructures 21 in contact with the light receiving surface (i.e., the bottom portions) may be composed of light-transmitting materials such as SiO2, while a surface of the microstructures 21 not in contact with the light receiving surface (i.e., the side portions) may be composed of opaque materials such as silicon. Alternatively, the bottom portions and the side portions of the microstructures 21 may both be composed of SiO2.
  • Alternatively, the microstructures 21 may be implemented in different embodiments to allow the carriers 22 to be more easily accommodated in the microstructures 21, taking into account the influence of air pressure or hydrodynamics on the carriers 22 during operation.
  • For example, FIG. 2 shows a biomolecule image sensor 100A according to another embodiment of the present invention. The microstructure layer 20A may be an array formed by a plurality of inverted pyramidal microstructures 21A, which facilitates the gathering of the incident light, and the inverted pyramidal microstructures 21A are preferably with 2.5 µm length and width. Alternatively, FIG. 3 shows a biomolecule image sensor 100B according to another embodiment of the present invention. The microstructure layer 20B may be an array formed by a plurality of honeycomb microstructures 21B. The most densely distributed without gaps between the honeycomb microstructures 21B is favorable for the carrier 22 to be evenly accommodated therein. The honeycomb microstructures 21B are preferably with 1.55 µm length and 2.5 µm depth.
  • FIGS. 4A and 4B show a biomolecule image sensor 200 according to another embodiment of the present invention. As shown in FIGS. 4A and 4B, in the biomolecular image sensor 200 of the present invention, the microstructure layer 40 may be an array formed by a plurality of microlenses 42, wherein a plurality of microstructures 41 are formed between the microlenses 42 to accommodate the biomolecule A or the carrier 22 carrying the biomolecule A. The microlenses 42 and the unit pixels 11 may be correspondingly arranged in different ways.
  • More specifically, as shown in FIG. 4A, on the unit pixels 11, four microlenses 42 may be arranged corresponding to one single unit pixel 11, wherein the area of the unit pixel 11 may be greater than or equal to the area surrounded by the four microlenses 42. Alternatively, as shown in FIG. 5 , in the biomolecule image sensor 200A of the present invention, the area of the unit pixel 11A may be smaller than the area surrounded by the four microlenses 42, and the microstructure 41 formed by the four microlenses 42 is arranged corresponding to the unit pixel 11A. Alternatively, as shown in FIG. 6 , in the biomolecule image sensor 200B of the present invention, each microlens 42 may be arranged corresponding to one single unit pixel 11B respectively.
  • As shown in FIGS. 7A and 7B, the method of detecting a biomolecule in a sample by the biomolecule image sensor of the present invention may be as following. A fluorescent marker or a chemiluminescent marker is added to the biomolecules in the sample through biological or chemical analysis such as ELISA, and the biomolecules are accommodated in the microstructures (S11). The incident light in one single microstructure is detected by the unit pixels respectively (S12), wherein the incident light includes the lights emitted by the fluorescent maker or the chemiluminescent marker of the biomolecule. Electrons are generated from the incident light received by each of the unit pixels through the photoelectric conversion element (S13). A voltage signal is generated according to the number of the electrons through the readout circuits (S14). The presence and/or concentration of the biomolecule are then analyzed according to the voltage signal.
  • Alternatively, when the biomolecule image sensor of the present invention is used to detect a biomolecule in a sample, the biomolecule containing fluorescent makers or chemiluminescent markers may also be placed on a substrate such as a glass slide, traditional biochips, or the like. The substrate is then placed on the biomolecule image sensor of the present invention, so that the unit pixels would detect an incident light from the corresponding position respectively. Then, electrons are also generated from the incident light through the photoelectric conversion element, and a voltage signal, which is used as the signal readout, is generated according to the number of the electrons through the readout circuits. The presence and/or concentration of the biomolecule are then analyzed according to the voltage signal.
  • When the biomolecule image sensor of the present invention is used to analyze the presence and/or concentration of the biomolecule, the quantification thereof may be not only performed by analog colorimetric method, that is, the incident light in one single microstructure received by the unit pixels is one single readout to determine the presence of the biomolecule (S151) or further compare with the standard curve for obtaining the concentration of the biomolecule (S161), but also by digital method, that is, according to the predetermined threshold (S152), the unit pixels with the signal readout exceeding the threshold are defined as 1 (S153) and the unit pixels with the signal readout not exceeding the threshold are defined as 0 (S154), and the number of unit pixels defined as 1 is calculated and compared with the standard curve (S162) to obtain the concentration of the biomolecule more accurately.
  • Further, traditional ELISA utilizes large and expensive equipment to receive optical or electrical signals after performing complex biochemical detection procedures to analyze the status of biochemical molecular reactions. The present invention is a completely innovative image sensor for detecting biomolecules prepared by a semiconductor process. The image sensor of the present invention, which is smaller than a coin, may directly detect the presence of specific biomolecules in samples and quantify concentration thereof, but does not require additional large equipment.
  • On the other hand, compared with the traditional biochip, the present invention does not require additional image capture system or equipment. That is, the biomolecular image sensor of the present invention provides the functions of biological or chemical analysis and image capture and interpretation. That is, the detection process may be directly operated on the biomolecule image sensor of the present invention, and corresponding detection results may be obtained in real time.
  • On the other hand, when quantifying biomolecular images by traditional methods, such as observation with a microscope, if the concentration of the biomolecules is extremely low, the biomolecules directly placed on the surface of the glass slide would distribute extremely uneven, resulting that only a small part of the area (i.e., the unit pixel) contains chemiluminescent or fluorescent signals, and therefore, the presence and intensity of the optical signals cannot be effectively interpreted within the overall field of view, so as to decrease the detection sensitivity. However, the biomolecule image sensor of the present invention independently detects the incident light in the corresponding microstructure through each unit pixel, and compares each measured signal readout with the thresholds independently, and therefore, even when the concentration of the biomolecules in the sample is extremely low, the presence and intensity of chemiluminescent or fluorescent signals of the biomolecules would still be accurately interpreted, so as to increase the detection sensitivity. Further, because the microstructure in the biomolecule image sensor of the present invention enables the biomolecules to be dispersed evenly on the light receiving surface, the photoelectric conversion element would easily determine the incident light really from the luminescent or fluorescent markers on the biomolecules. The biomolecule image sensor of the present invention provides the quantitative mode of analog colorimetric method and digital method, and could be switched according to different concentrations of biomolecules, so as to maximize the detection range.

Claims (15)

What is claimed is:
1. A biomolecular image sensor, comprising:
an image sensing element, containing a plurality of unit pixels disposed in an array on a substrate, wherein each of the plurality of unit pixels contains at least one photoelectric conversion element, the photoelectric conversion element receives an incident light to generate electrons, and a surface of the image sensing element receiving the incident light is defined as a light receiving surface; and
a microstructure layer, disposed on the light receiving surface of the image sensing element and having a plurality of microstructures arranged in a specific shape repeatedly, wherein each of the plurality of microstructures corresponds to at least one unit pixel.
2. The biomolecular image sensor according to claim 1, wherein the microstructure layer is an array formed by a plurality of inverted pyramidal or honeycomb microstructures.
3. The biomolecular image sensor according to claim 1, wherein each of the microstructures is formed on the image sensor element by photolithography process or imprint lithography process.
4. The biomolecular image sensor according to claims 1, wherein each of the microstructures is provided to accommodate at least one biomolecule.
5. The biomolecular image sensor according to claim 4, wherein the incident light is a light emitted by a fluorescent marker or a chemiluminescent marker on the biomolecule.
6. The biomolecular image sensor according to claim 5, further comprising a carrier carrying the biomolecule, and the carrier is accommodated in the microstructure.
7. The biomolecular image sensor according to claim 6, wherein the carrier is a microparticle.
8. A method of detecting a biomolecule, comprising:
(a) providing the biomolecular image sensor according to claims 1;
(b) accommodating the biomolecule with a fluorescent marker or a chemiluminescent marker in each of the plurality of microstructures;
(c) detecting an incident light in each of the plurality of microstructures by each of the plurality of unit pixels, wherein the incident light comprises a light emitted by the fluorescent marker or the chemiluminescent marker;
(d) generating electrons from the incident light detected by each of the plurality of unit pixels by the photoelectric conversion element;
(e) generating a voltage signal based on a number of the electrons by a readout circuit coupled to each of the plurality of unit pixels; and
(f) analyzing a concentration of the biomolecule based on the voltage signal.
9. A biomolecular image sensor, comprising:
an image sensing element, containing a plurality of unit pixels disposed in an array on a substrate, wherein each of the plurality of unit pixels contains at least one photoelectric conversion element, the photoelectric conversion element receives an incident light to generate electrons, and a surface of the image sensing element receiving the incident light is defined as a light receiving surface; and
a microstructure layer, disposed on the light receiving surface of the image sensing element and being an array formed by a plurality of microlenses, wherein a plurality of microstructures are formed between the plurality of microlenses, and each of the plurality of microstructures corresponds to at least one unit pixel.
10. The biomolecular image sensor according to claim 9, wherein each of the plurality of microlenses is formed on the image sensor element by photolithography process or imprint lithography process.
11. The biomolecular image sensor according to claims 9, wherein each of the microstructures is provided to accommodate at least one biomolecule.
12. The biomolecular image sensor according to claim 11, wherein the incident light is a light emitted by a fluorescent marker or a chemiluminescent marker on the biomolecule.
13. The biomolecular image sensor according to claim 12, further comprising a carrier carrying the biomolecule, and the carrier is accommodated in the microstructure.
14. The biomolecular image sensor according to claim 13, wherein the carrier is a microparticle.
15. A method of detecting a biomolecule, comprising:
(a) providing the biomolecular image sensor according to claims 9;
(b) accommodating the biomolecule in a sample in each of the plurality of microstructures;
(c) detecting an incident light in each of the plurality of microstructures by each of the plurality of unit pixels;
(d) generating electrons from the incident light detected by each of the plurality of unit pixels by the photoelectric conversion element;
(e) generating a voltage signal based on a number of the electrons by a readout circuit coupled to each of the plurality of unit pixels; and
(f) analyzing a presence and/or a concentration of the biomolecule based on the voltage signal;
wherein, a fluorescent marker or a chemiluminescent marker is added on the biomolecule before or after (b), and the incident light comprises a light emitted by the fluorescent marker or the chemiluminescent marker.
US17/731,840 2021-10-14 2022-04-28 Biomolecular image sensor and method thereof for detecting biomolecule Pending US20230119978A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US17/731,840 US20230119978A1 (en) 2021-10-14 2022-04-28 Biomolecular image sensor and method thereof for detecting biomolecule

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202163255446P 2021-10-14 2021-10-14
US17/731,840 US20230119978A1 (en) 2021-10-14 2022-04-28 Biomolecular image sensor and method thereof for detecting biomolecule

Publications (1)

Publication Number Publication Date
US20230119978A1 true US20230119978A1 (en) 2023-04-20

Family

ID=85961153

Family Applications (3)

Application Number Title Priority Date Filing Date
US17/731,919 Pending US20230123442A1 (en) 2021-10-14 2022-04-28 Biomolecular image sensor and method thereof for detecting biomolecule
US17/731,840 Pending US20230119978A1 (en) 2021-10-14 2022-04-28 Biomolecular image sensor and method thereof for detecting biomolecule
US17/739,230 Pending US20230123106A1 (en) 2021-10-14 2022-05-09 Bioassay device and bioassay system comprising the same

Family Applications Before (1)

Application Number Title Priority Date Filing Date
US17/731,919 Pending US20230123442A1 (en) 2021-10-14 2022-04-28 Biomolecular image sensor and method thereof for detecting biomolecule

Family Applications After (1)

Application Number Title Priority Date Filing Date
US17/739,230 Pending US20230123106A1 (en) 2021-10-14 2022-05-09 Bioassay device and bioassay system comprising the same

Country Status (3)

Country Link
US (3) US20230123442A1 (en)
CN (3) CN115980022A (en)
TW (3) TWI825609B (en)

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140274747A1 (en) * 2013-03-15 2014-09-18 Illumina, Inc. Super resolution imaging
CN103604784B (en) * 2013-11-05 2016-05-18 浙江工业大学 Array sensing chip is analyzed in the fluoroscopic examination of CMOS contact
JP6461196B2 (en) * 2014-12-26 2019-01-30 株式会社東芝 Biosensor
CN105543071A (en) * 2015-12-10 2016-05-04 严媚 CMOS and ISFET dual-mode image chemical sensor chip for high-throughput gene sequencing
SG11201908509TA (en) * 2017-03-20 2019-10-30 Mgi Tech Co Ltd Biosensors for biological or chemical analysis and methods of manufacturing the same
NL2020621B1 (en) * 2018-01-08 2019-07-15 Illumina Inc Multiplexing of an active sensor detector using structured illumination
US10670526B2 (en) * 2018-03-05 2020-06-02 Smartsens Technology (Cayman) Co., Limited DNA sequencing system with stacked BSI global shutter image sensor

Also Published As

Publication number Publication date
US20230123442A1 (en) 2023-04-20
TWI825609B (en) 2023-12-11
TW202326102A (en) 2023-07-01
US20230123106A1 (en) 2023-04-20
CN115980023A (en) 2023-04-18
CN115980024A (en) 2023-04-18
TW202317969A (en) 2023-05-01
TW202331227A (en) 2023-08-01
TWI821935B (en) 2023-11-11
CN115980022A (en) 2023-04-18

Similar Documents

Publication Publication Date Title
US11112338B2 (en) Dual-image based bioimaging devices and techniques
EP3492915B1 (en) Optical sensor for analyte detection
CN102165305B (en) Detection system and method
Obahiagbon et al. A compact, low-cost, quantitative and multiplexed fluorescence detection platform for point-of-care applications
US11112347B2 (en) Classifying microbeads in near-field imaging
US20100253323A1 (en) Magnetic washing for biosensor
US20100243914A1 (en) Super critical angle fluorescence scanning system
US20230119978A1 (en) Biomolecular image sensor and method thereof for detecting biomolecule
Kurzbuch et al. A biochip reader using super critical angle fluorescence
Lin et al. Fluorescence measurements, imaging and counting by a smartphone
KR102347135B1 (en) Device and methods for detecting norovirus using time-resolved fluorescence
US20130323855A1 (en) Device for holding a sample
US10633734B2 (en) Optical sensor for analyte detection
JP6228206B2 (en) Protein-specific light detection
Kishore et al. Fabrication of a low-cost benchtop optical imager for quantum dot microarray-based stress biomarker detection
Obahiagbon Modeling, Design, Fabrication, and Characterization of a Highly Sensitive Fluorescence-based Detection Platform for Point-of-Care Applications
WO2023086794A1 (en) Systems and methods for digital affinity-based detection assays
US20200056995A1 (en) Biological substance analysis method, biological substance analyzer, biological substance analysis program, and biological substance analysis system
Bell et al. An integrated digital imaging system and microarray mapping software for rapid multiplexed quantitation of protein microarray immunoassays

Legal Events

Date Code Title Description
AS Assignment

Owner name: GUANGZHOU TYRAFOS SEMICONDUCTOR TECHNOLOGIES CO., LTD., CHINA

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:YIN, PING-HUNG;SHAO, CHUN-I;SUN, HSIAO-WEN;AND OTHERS;REEL/FRAME:059768/0309

Effective date: 20220426

STPP Information on status: patent application and granting procedure in general

Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION