WO2012125906A1 - Appareil et procédé pour l'analyse de données de puces à cellules - Google Patents

Appareil et procédé pour l'analyse de données de puces à cellules Download PDF

Info

Publication number
WO2012125906A1
WO2012125906A1 PCT/US2012/029391 US2012029391W WO2012125906A1 WO 2012125906 A1 WO2012125906 A1 WO 2012125906A1 US 2012029391 W US2012029391 W US 2012029391W WO 2012125906 A1 WO2012125906 A1 WO 2012125906A1
Authority
WO
WIPO (PCT)
Prior art keywords
fluorescent
module
data
spots
curve
Prior art date
Application number
PCT/US2012/029391
Other languages
English (en)
Inventor
Sang Youl JEON
Bo Sung Ku
Sreekanth Karuvelil RAMACHANDRAN
Moo-Yeal Lee
Original Assignee
Solidus Biosciences, Inc.
Samsung Electro-Mechanical 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 Solidus Biosciences, Inc., Samsung Electro-Mechanical Co, Ltd filed Critical Solidus Biosciences, Inc.
Publication of WO2012125906A1 publication Critical patent/WO2012125906A1/fr

Links

Classifications

    • 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
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06V—IMAGE OR VIDEO RECOGNITION OR UNDERSTANDING
    • G06V20/00—Scenes; Scene-specific elements
    • G06V20/60—Type of objects
    • G06V20/69—Microscopic objects, e.g. biological cells or cellular parts

Definitions

  • the present invention relates to an apparatus and method for analyzing data of cell chips.
  • cell chips refers to chips in which living or resting cells are placed or maintained on substrates, preferably glass substrates, e.g. under conditions suitable for conducting one or more cellular reactions.
  • Such cell chips are used in the pharmaceutical field or the cosmetic-related field to test the reactions of cells to chemicals in order to assess the biological activity, efficacy and/or safety of the chemicals.
  • Examples include the cell chips described in WO 2007/053561 andUSSN 61/381,812, filed on September 10, 2010, for example, which is incorporated herein by reference in its entirety.
  • test chemicals are applied to an enzyme- containing chip that carries out enzymatic reactions.
  • enzyme-containing chips include the chips described in US Patent 7,267,958 andUSSN 61/381,812, filed on September 10, 2010, for example, which is incorporated herein by reference in its entirety.
  • the enzyme-containing chip is contacted with the cell chip so that the test compound or test compound that is enzymatically reacted is applied to cells.
  • the cells are dyed with fluorescent material to facilitate analysis of the reaction of the test compound or enzymatically reacted test compound with the cells.
  • the conditions of the cells are investigated and analyzed by test devices, such as scanners, etc., to determine whether the chemicals are biologically active or toxic.
  • the present invention has been made to provide an apparatus for automatically analyzing data obtained from one or more cell chips, thus simplifying the test process, reducing the required time, and enhancing the precision of the testing process, and a method of analyzing data of cell chips using the apparatus.
  • a cell chip scanner scans one or more cell chips and creates image files thereof.
  • the cell chips have preferably been subjected to a compound, which upon contact with a cell or a component thereof (e.g., ATP), fluoresces.
  • a fluorescent-intensity measuring module measures fluorescent intensities of the spots in the image files.
  • An analyzer creates graphs in such a way as to individually conduct curve-fitting with respect to the fluorescent intensities of the spots.
  • the analyzer creates an integrated graph in such a way as to integrate pieces of data of the blocks and then conduct curve- fitting on the integrated data.
  • the apparatus may further include a computer which can accept, analyze, store, and display data provided by the operator relating to the cell chip, the images created by the scanner, and the results created by the analyzer.
  • the apparatus can include a storage module for storing information relating to the cell chips and/or test conditions.
  • a display may provide a data, or test condition, input window to a tester or operator.
  • a control module may provide the test condition input window on the display.
  • the tester or operator can input data or test conditions relating to the cell chip with an input module, the data then being stored in the storage module.
  • the analyzer may create the graphs in such a way as to individually conduct the curve-fitting with respect to the blocks while referring to the test conditions stored in the storage module.
  • the analyzer may include an individual curve-fitting and graph-creating module and an integrated curve-fitting and graph-creating module.
  • the individual curve-fitting and graph- creating module may create the graphs in such a way as to individually conduct the curve-fitting with respect to the fluorescent intensities of the spots of the blocks that are measured by the fluorescent-intensity measuring module. For example, where certain spots within a block (a line of spots, for example) differ from one another in test compound concentration (a progression of dilutions of a test compound contacted with the spots, for example), the individual curve-fitting and graph-creating module can create a graph that permits analysis of the fluorescence intensity of those spots as a function of concentration.
  • the integrated curve-fitting and graph-creating module may create the integrated graph in such a way as to integrate the pieces of data of the fluorescent intensities of the spots of the blocks that are measured by the fluorescent-intensity measuring module under the same test conditions and then conduct curve-fitting on the integrated data.
  • One or both of these modules can be incorporated into a single software program to be run by the computer or control module discussed above.
  • the data received from the operator or tester can be used to identify spots or compositions thereof.
  • the individual curve-fitting and graph-creating module may include a data selection unit selecting a plurality of fluorescent intensities that were obtained by conducting repetitive tests or replicates in a block or chip.
  • a constant determining unit may determine a constant of a curve equation using the test results.
  • a graph creating unit may create the graph using the constant determined by the constant determining unit.
  • the software can solve the equation for a specific value (e.g., an EC50 or IC50) and, optionally, incorporate that solution into a graph, report or other results.
  • the analyzer may optionally include an integrated curve-fitting and graph-creating module may include a data integrating unit integrating data obtained from a plurality of cell chips.
  • a constant deteimining unit may deteimine a constant of a curve equation using the integrated data.
  • a graph creating unit may create the integrated graph.
  • the analyzer may include an error data control module and a report creation module.
  • the error data control module may classify the data and identify an item to be checked by the tester or operator and, if appropriate, permit the tester to delete the item or erroneous data.
  • the report creation module may classify a result of the curve-fitting with respect to the integrated data and the scanned images, and create a result report, and then output the result report.
  • the fluorescent-intensity measuring module may include a spot position detecting unit detecting positions of the spots in such a way as to calculate an average of Y-axial fluorescent- intensities and an average of X-axial fluorescent-intensities from the image files created by scanning the cell chips using the cell chip scanner.
  • a spot boundary deteimining unit may deteimine boundaries of the spots detected by the spot position detecting unit.
  • a spot fluorescent- intensity calculating unit may calculate a fluorescent-intensity of each of the spots after the boundaries of the spots are deteimined by the spot boundary deteimining unit.
  • image files are created by scanning a plurality of cell chips using a cell chip scanner. Fluorescent intensities of spots are measured by a fluorescent-intensity measuring module in the image files created by the cell chip scanner. Graphs are created in such a way as to individually conduct curve-fitting with respect to the fluorescent intensities of the spots of blocks that are measured by the fluorescent-intensity measuring module. An integrated graph can be created in such a way as to integrate pieces of data that are related to the fluorescent intensities of the spots that are measured by the fluorescent-intensity measuring module under same test conditions and then conduct curve-fitting with respect to the integrated data.
  • Each of the cell chips may include blocks. Each of the blocks may have a plurality of spots.
  • the spots can preferably contain cells and one or more compounds to be tested.
  • the compounds added to each spot can be the same or different.
  • the spots within a block contain the same compound at different concentrations or dilutions.
  • the compounds are contacted with an enzyme or enzymes for reaction (e.g., metabolism) and the reaction products (e.g., metabolites) of the compound can then be contacted with the cells in the cell chip.
  • the cells in the cell chip can be contacted with a fluorescent marker that detects, for example, cytotoxicity.
  • an operator or tester can input data or information relating to the chip and relevant to the test into the apparatus.
  • a display module can present a test condition input window to a tester, where the tester, using an input module, can input data or information for storage in a storage module.
  • integrated results may be classified by an error data control module as above.
  • positions of the spots may be detected.
  • the apparatus calculates an average of Y-axial fluorescent-intensities and an average of X-axial fluorescent-intensities from the image files. Boundaries of the spots may be determined by the fluorescent-intensity measuring module. A fluorescent-intensity value for each spot may be calculated after the boundaries of the spots are determined by the fluorescent-intensity measuring module.
  • pixels of a predetermined value or more may be connected to each other with lines (or line segments) using the fluorescent-intensity measuring module.
  • the lines are generated in a specific orientation (e.g., horizontal or vertical, relevant to a boundary of the chip or a block).
  • the lines can be generated in a parallel fashion of a predetermined distance, x, wherein x is less than the predicted radius of a spot and preferably less than 20%, such as less than 10%> of the predicted radius of a spot.
  • Lines may be united, thereby forming a longer line (or line segment), when a distance between the lines is less than a predetermined value, for example, distance y, wherein y is less than the predicted radius of a spot and preferably less than 20%>, such as less than 10%> of the predicted radius of a spot.
  • a line may be deleted when a length of the line is shorter than a predetermined value, for example distance z, wherein z is less than the predicted radius of a spot and preferably less than 20%>, such as less than 10%> of the predicted radius of a spot.
  • One or more lines (but preferably less than a majority of the total number of lines drawn) that are spaced apart from a plurality of other lines by a predetermined distance or more (for example, distance v, wherein v is greater than 10%>, such as greater than 20% of the predicted radius of the spot) may be deleted.
  • Midpoints of the lines may be determined and the central midpoint may be determined.
  • the boundary of the spot is calculated using a predetermined radius (e.g., the predicted radius).
  • an average of absolute intensities of pixels in a spot area may be determined.
  • a central value of pixels in a peripheral area e.g., an area outside the boundary of any spot
  • the central value of the pixels of the peripheral area may be subtracted from the average of the absolute intensities of the pixels of the spot area, thus attaining the fluorescent- intensity of the spot.
  • a plurality of values of fluorescent intensities that are results of various spots on a chip or within a block or repeated tests may be selected by an individual or integrated curve-fitting and graph-creating module.
  • a constant of a curve equation may be determined and/or one or more graphs may be created.
  • FIG. 1 is a diagram showing the construction of an apparatus for analyzing data of cell chips, according to an embodiment of the present invention
  • FIG. 2 is a view showing one example of a cell chip used in the present invention
  • FIG. 3 illustrates one example of a test condition input window used in the present invention
  • FIG. 4 is a view illustrating a process of detecting spots using a fluorescent-intensity measuring module according to the present invention
  • FIGS. 5A, 5B and 6 through 10 are views illustrating a process of determining a boundary of a spot using the fluorescent-intensity measuring module according to the present invention
  • FIG. 11 is a view illustrating a process of measuring fluorescent-intensities of the spots using the fluorescent-intensity measuring module according to the present invention.
  • FIG. 12 is a table of data related to the fluorescent-intensities of one block of an individual cell chip which are obtained by the fluorescent-intensity measuring module according to the present invention.
  • FIG. 13 is a graph showing the results of testing four kinds of enzymes with respect to the same compound created in an individual curve-fitting and graph-creating module according to the present invention.
  • FIG. 14 is a view showing one example of an image used for sorting out items that can be examined using an error data control module according to the present invention.
  • FIG. 15 is an example showing an image including scan images and curve-fitting graphs that illustrate in detail the examination-required item sorted out by the error data control module according to the present invention
  • FIG. 16 is a view showing one example of a report created by a report creation module according to the present invention
  • FIG. 17 is a flowchart of a method of analyzing data of cell chips, according to an embodiment of the present invention.
  • FIG. 18 is a flowchart of a test condition input step of FIG. 17;
  • FIG. 19 is a flowchart of a spot position detecting step of FIG. 17;
  • FIG. 20 is a flowchart of a spot fluorescent-intensity measuring step of FIG. 17.
  • FIG. 21 is a flowchart of an individual curve-fitting and graph creating step of FIG. 17.
  • FIG. 1 is a diagram showing the construction of an apparatus of analyzing data of cell chips, according to an embodiment of the present invention.
  • the cell-chip data analysis apparatus (hereinafter, referred to as "data analysis apparatus") according to the embodiment of the present invention includes a cell chip scanner 100, a control module 200, a display module 300, an input module 400, a storage module 500, a fluorescent-intensity measuring module 600 and an analyzer 700.
  • the analyzer 700 includes an individual curve-fitting and graph-creating module 710, an error data control module 720, an integrated curve-fitting and graph-creating module 730 and a report creation module 740.
  • the fluorescent-intensity measuring module 600 includes a spot position detecting unit 610, a spot boundary determining unit 620 and a spot fluorescent-intensity calculating unit 630.
  • the individual curve-fitting and graph-creating module 710 of the analyzer 700 includes a data selection unit 711 , a constant determining unit 712 and a graph creating unit 713.
  • the integrated curve-fitting and graph-creating module 730 of the analyzer 700 includes a data integrating unit 731 , a constant determining unit 732 and a graph creating unit 733.
  • the cell chips receive, for example, from enzyme-containing chips, substances, including products created by reacting enzymes with chemicals in the enzyme- containing chips.
  • the cells on the cell chips can then respond to the substances for a toxic reaction or a biologically active outcome.
  • a toxic reaction can be cell death or apoptosis.
  • the cell chips can be dyed with a fluorescent material.
  • the cell chip scanner 100 then scans the cell chip(s).
  • the chip contains enzymes, such as liver enzymes (e.g., human or other animal), placed on a glass substrate under conditions suitable for enzymatic activity.
  • enzymes such as liver enzymes (e.g., human or other animal)
  • the enzymes can react with the test chemicals and form a product, such as a metabolite.
  • the cell chip incorporates human cells, including cells isolated from a human body on a substrate.
  • Cells that can be used, or the tissues/organs they can be derived from include, but are not limited to bone marrow, skin, cartilage, tendon, bone, muscle (including cardiac muscle), blood vessels, corneal, neural, brain, gastrointestinal, renal, liver, pancreatic (including islet cells), lung, pituitary, thyroid, adrenal, lymphatic, salivary, ovarian, testicular, cervical, bladder, endometrial, prostate, vulval, esophageal, etc.
  • the various cells of the immune system such as T lymphocytes, B lymphocytes, polymorphonuclear leukocytes, macrophages, and dendritic cells.
  • the cells are normal cells.
  • the cells can be cancerous, derived from a cell line or an immortalized cell.
  • the cells can be bacterial, yeast or derived from a plant.
  • the cells preferably are placed on a substrate.
  • substrates include glass substrates.
  • the cells can be encapsulated into or layered over a matrix that can, for example, support growth.
  • Such matrices include alginate or collagen and can include an appropriate cell culture media.
  • the matrices and/or cells can be spotted or placed onto a flat substrate, into wells or upon columns.
  • the cells are placed on the substrate in discrete "spots". The spots can be deemed to be discrete if the spots are not touching, in fluid communication or are physically separated.
  • the cell chip can preferably be organized as a two dimensional array having a plurality of rows and columns.
  • Each cell chip can include a plurality of "blocks."
  • a block is a subarray, section or portion of the array.
  • Each block includes a plurality of spots, preferably arranged in a plurality of columns and rows.
  • the spots within a block are the same in composition, differing one from another only in the concentrations of one or more components, such as the substance being tested (e.g., a test chemical, enzymatic reaction product or metabolite).
  • the spots within each block can possess the same composition, differing one from another block only in one component of the composition, such as the substance being tested, an enzyme or the cell. For example, as shown in FIG.
  • the cell chip includes 24 blocks which are formed by a combination of four kinds of enzymes (for the sake of convenience, referred to as A, B, C and D) and six kinds of compounds (for the sake of convenience, referred to as 1 , 2, 3, 4, 5 and 6).
  • the cell can be a constant in all blocks.
  • Alternative configurations of the components can be envisioned and made by the person of skill in the art.
  • each block includes a matrix with four rows having the same compound concentration and seven columns having different enzyme concentrations (e.g., having dilutions of one-fold, two-fold, three-fold, four-fold, five-fold, six-fold and seven-fold).
  • the four rows thereby, provide four replicates of the experiment.
  • One or more cell chips are scanned in sequence by a scanner, such as the cell chip scanner 100.
  • the results of the scanning can be stored as image files (for example, TIFF files, PDF files, BMP files, etc.) in the storage module 500, which is generally a computer or CPU.
  • the control module 200 controls the operation of the parts of the data analysis apparatus.
  • control module 200 shows the test condition input window on the display module 300 such that a tester (or operator) can input test conditions using the input module 400, which can be, for example, a keyboard.
  • the control module 200 stores test conditions input by the tester in the storage module 500.
  • the display module 300 can display one or more of the image files scanned by the cell chip scanner 100, the test condition input window provided by the control module 200, a table showing fluorescent intensities measured by the fluorescent-intensity measuring module 600, a graph created by the individual curve-fitting and graph-creating module 710 of the analyzer 700, a target item sorted out by the error data control module 720 of the analyzer 700, an integrated graph created by the integrated curve-fitting and graph-creating module 730 of the analyzer 700, or a report created by the report creation module 740 of the analyzer 700.
  • the test conditions are entered, which can include a project folder title, a chip platform (for example, "672 spot layout"), the number of compound dilution folds (for example, the "4"), a cell type (for example, "Hep3B") relative to each of slide names (for example, stated as “2010-07- 14-M” and “2010-07- 14- "), a unit (for example, a "micro mole”), the compound identifiers (for example, compound designations "AA”, “BB”, “CC”, “DD”, “EE”, “FF”, “GG”, “HH”, “ ⁇ ", “JJ”, “KK” and “LL”), and the enzymes (for example, designated as “control”, “P450”, “Phase ⁇ ” and "All mix”).
  • a project folder title for example, a chip platform (for example, "672 spot layout")
  • the number of compound dilution folds for example, the "4"
  • a cell type for example, "Hep3B” relative to each of slide names (
  • the fluorescent-intensity measuring module 600 detects spots in the image files created by scanning the cell chips using the cell chip scanner 100, deteimines boundaries of the spots and then calculates fluorescent-intensities of the spots.
  • the spot position detecting unit 610 of the fluorescent- intensity measuring module 600 calculates averages of X-axial fluorescent-intensities and averages of Y-axial fluorescent-intensities from the image files created by scanning the cell chips using the cell chip scanner 100. Thereafter, the spot position detecting unit 610 determines junctions of the maximums of the averages of X-axial fluorescent-intensities and the maximums of the averages of Y-axial fluorescent-intensities as approximate positions of the respective spots. With regard to this, FIG.
  • FIG. 4 illustrates a process of determining a junction of a first maximum of the X-axial fluorescent-intensity averages and a first maximum of the Y-axial fluorescent-intensity averages as an approximate position of a first spot using the spot position detecting unit 610.
  • the spot boundary determining unit 620 connects pixels of a predetermined value or more (e.g. , greater than 30%, preferably greater than 50% of the average of absolute intensities of the pixels in the spot area) to each other with lines.
  • FIG. 5B removes the pixels and retains the drawn lines.
  • the lines are united.
  • FIG. 7 when a length of a line is shorter than a predetermined value, the line is deleted.
  • FIG. 8 a minor number of lines which are spaced apart from the other lines by a predetermined distance or more are deleted.
  • midpoints of the lines are determined.
  • an average of the midpoints of the lines is determined and then the average is determined as the center of the spot.
  • the boundary can then be determined by using a predetermined radius measured from the center of the spot.
  • the spot fluorescent-intensity calculating unit 630 of the fluorescent-intensity measuring module 600 can calculate the fluorescent-intensity of the spots.
  • the spot fluorescent-intensity calculating unit 630 determines the average of absolute intensities of pixels in a spot area (e.g., within the boundary of the spot).
  • the unit can, optionally, determine a central value of pixels in a peripheral area (e.g., outside the boundary of the spots), and subtract the central value of the pixels of the peripheral area from the average of the absolute intensities of the pixels of the spot area. Thereby, the fluorescent- intensity of the spot is attained.
  • the analyzer 700 creates graphs to individually conduct curve-fitting for the fluorescent intensities.
  • the analyzer 700 can also create an integrated graph with data from a plurality cell chips.
  • the analyzer 700 can additional create and output one or more reports.
  • the analyzer 700 can identify potentially erroneous data and enable the tester to delete erroneous data.
  • the individual curve-fitting and graph-creating module 710 of the analyzer 700 conducts curve-fitting on the fluorescent intensity of selected spots within each block of the cell chip. For example, spots within a block that differ one from another in the concentrations of a test compound or metabolite can potentially result in differing fluorescent intensities as a function of concentration.
  • the module can analyze the data create a graph resulting from curve-fitting.
  • the data selection unit 711 of the individual curve-fitting and graph-creating module 710 can select a plurality of fluorescent intensities that have been obtained by repetitive tests with respect to concentrations of compounds of a plurality of blocks.
  • the table of FIG. 12 provides fluorescent intensities resulting from four repetitions with respect to seven dilutions of a compound.
  • the data selection unit 711 selects the values resulting from the repeated tests.
  • the constant determining unit 712 can determine constants (e.g., d, g and b) of a curve equation (e.g., Equation 1) for example, using numerical analysis.
  • y denotes fluorescent intensity
  • x denotes the number of dilution folds (log concentration)
  • g denotes an inflection point at which the survival rate of a cell becomes 50% (that is, the value of g denotes IC50).
  • FIG. 13 illustrates curve graphs showing results of individual curve-fitting processes with respect to four kinds of enzymes A, B, C and D.
  • Square marks denote inflection points at which the survival rates of cells become 50%.
  • the error data control module 720 of the analyzer 700 classifies the result, and if items required to be checked by the tester are detected, the error data control module 720 sorts out the items so as to enable the tester to delete erroneous data.
  • the error data control module 720 can indicate this and cause the data to be displayed and present remove buttons on the display module 300.
  • the actuation of the remove button can cause the data to be deleted from the storage unit and/or from data integration.
  • FIG. 14 shows the case where with regard to the compound AA and the enzyme agent ED of the slide name 2010-07- 14-M.tif, the compound BB and the enzyme agent EC of the slide name 2010-07-M.tif, and the compound CC and the enzyme agent EC of the slide name 2010-07- 14-M.tif, checkboxes are checked to notify the tester if it has to be checked whether there is data to be excluded or deleted.
  • the error data control module 720 provides the scanned images and the curve-fitting graphs to enable the tester to check in detail whether there is a data error.
  • the integrated curve-fitting and graph-creating module 730 of the analyzer 700 integrates data from the cell chips (for example, repeat tests) into a single piece of data, and determines constants of the curve equation, and creates an integrated graph, as described above.
  • the report creation module 740 classifies the results of curve-fitting with respect to the integrated data and the scanned images and outputs a result report.
  • the result report created by the report creation module 740 is preferably editable.
  • An example of a report is illustrated in FIG. 16. This exemplary report includes a summary, the scanned images and an integrated curve-fitting graph.
  • FIG. 17 is a flowchart of a method of analyzing the data of cell chips, according to an embodiment of the present invention.
  • the method includes a cell chip scanning step S10, a test condition input step S20, a spot position detecting step S30, a spot fluorescent-intensity measuring step S40, an individual curve-fitting and graph creating step S50, a test result classifying and abnormal item sorting-out step S60, a data error item checking and deletion target data selection step S70, an integrated data curve-fitting and graph creating step S80 and a test result report creating step S90.
  • the scanner scans a cell chip that (1) receives, for example, from an enzyme-containing chip, a product (e.g., a metabolite) created by reacting one or more enzymes with chemicals added to the enzyme-containing chip, and (2) carries out a cellular reaction with the product to test for cytotoxicity, and is dyed with fluorescent material to detect cytotoxicity.
  • a product e.g., a metabolite
  • the cell chip scanner scans other cell chips in sequence by repeating the above-mentioned process and stores results of the scanning as image files.
  • the control module shows the test condition input window on the display module and the tester appoints a project folder, at step S21.
  • the tester inputs a chip platform.
  • the tester inputs the dilutions of the compound or chemical.
  • the tester inputs a cell type and other cell information.
  • the tester inputs compound information (e.g., compound identifiers).
  • the tester inputs enzyme information.
  • the fluorescent-intensity measuring module loads images from the project folder, at step S31.
  • the fluorescent-intensity measuring module detects an approximate position of a first spot from the loaded image.
  • a precise position of the first spot is deteimined in a predeteimined section centered on the detected approximate position.
  • the fluorescent-intensity measuring module uses averages of X-axial fluorescent-intensities and averages of Y-axial fluorescent- intensities.
  • Deteimining the precise position of a first spot on the chip, block or predeteimined section centered on a detected approximate position can include one or more of the following steps: connecting pixels of a predetermined value or more to each other with lines, uniting lines the distance between which is less than a predetermined value, deleting lines that are shorter than a predetermined value, deleting a minor number of lines spaced apart from the other lines by a predetermined distance or more, attaining midpoints of the remaining lines, and then averaging the midpoints of the lines. Additional spots can optionally be detected by these methods or by calculation, using the predicted distance between spots, or be a combination thereof.
  • the fluorescent-intensity measuring module determines whether the positions of all spots have been detected. If module detects another spot or concludes that the positions of all the spots have still not been detected, an approximate position of a subsequent spot is detected and the steps are repeated from step S33.
  • the fluorescent-intensity measuring module deteimines that the positions of all the spots have been detected, the center points of the spots are stored, at step S36. At step S37, it is determined whether all the images have been processed. If the module determines that another image is available for processing, the above steps can be repeated.
  • the fluorescent- intensity measuring module loads the images, at step S41.
  • the fluorescent-intensity measuring module attains an average of absolute intensities of pixels in each spot area.
  • the fluorescent-intensity measuring module attains a central value of pixels in each peripheral area.
  • the fluorescent-intensity measuring module subtracts the central value of the pixels of the peripheral area from the average of the absolute intensities of the pixels of the spot area, thus attaining the fluorescent-intensity of the spot.
  • the fluorescent-intensity measuring module determines whether all the spots have been processed. At step S46, if all the spots have still not been processed, the fluorescent- intensity measuring module can process the subsequent spot. At step S47, when all the spots have been processed, the fluorescent-intensity measuring module determines whether all the images are processed. At step S48, the fluorescent-intensity measuring module stores the fluorescent- intensities of the spots.
  • the individual curve-fitting and graph-creating module loads information regarding the fluorescent- intensities of the spots, at step S51.
  • the individual curve-fitting and graph-creating module selects data, for example, relating to the compound dilutions or concentrations (x) and the fluorescent-intensity (y) with respect to one test condition, e.g. residing in one block.
  • the constants d, g and b of the curve equation are determined.
  • the individual curve-fitting and graph-creating module conducts numerical analysis, during which it can calculate a numerical formula error with respect to the selected constants d, g and b of the curve equation (at step S54); and, determines whether the error is less than a reference value (at step S54); and step S54 is repeated if the error is not less than the reference value.
  • step S56 where the numerical analysis reveals that the error is less than the reference value, the individual curve-fitting and graph-creating module can determine whether all the test conditions (all the blocks) have been processed and repeat the above steps if necessary or desired.
  • the constants d, g and b can be stored and a graph is created and displayed or printed, at step S57.
  • the error data control module classifies the results of the tests and sorts out items for checking by the tester.
  • the error data control module indicates this and, optionally, displays a remove button or other method for removing or deleting data on the display module 300.
  • the error data control module can also display the scanned images and the curve-fitting graphs to facilitate whether the data should be excluded.
  • the tester checks target items sorted out by the error data control module.
  • the tester can remove the data by, for example, selecting a remove button for the corresponding item.
  • the integrated curve-fitting and graph-creating module integrates data of the cell chips, deteimines constants of the curve equation, and creates an integrated graph, as described above, and, if desired, a report of the results.
  • the report is optionally editable.
  • the apparatus according to the present invention automatically, efficiently and quickly processes the results of testing cell chips with improved accuracy.

Landscapes

  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Biochemistry (AREA)
  • Analytical Chemistry (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Multimedia (AREA)
  • Theoretical Computer Science (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

La présente invention concerne un appareil et un procédé pour l'analyse de données de puces à cellules. L'appareil comporte un scanneur de puces à cellules, un module de mesure d'intensité fluorescente et un analyseur. Le scanneur de puces de cellules effectue un balayage des puces à cellules et crée des fichiers d'images. Le module de mesure d'intensité de fluorescence mesure des intensités de fluorescence des taches dans les fichiers d'images créés par le scanneur de puces à cellules. L'analyseur d'images crée des graphiques de manière à effectuer individuellement un ajustement de courbes sur les intensités de fluorescence des taches des blocs qui ont été mesurés par le module de mesure d'intensité de fluorescence. L'analyseur crée un graphique intégré de manière à intégrer des pièces de données des blocs obtenus dans des mêmes conditions de test et ensuite effectue un ajustement de courbes sur les données intégrées.
PCT/US2012/029391 2011-03-16 2012-03-16 Appareil et procédé pour l'analyse de données de puces à cellules WO2012125906A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201161453241P 2011-03-16 2011-03-16
US61/453,241 2011-03-16

Publications (1)

Publication Number Publication Date
WO2012125906A1 true WO2012125906A1 (fr) 2012-09-20

Family

ID=46831111

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2012/029391 WO2012125906A1 (fr) 2011-03-16 2012-03-16 Appareil et procédé pour l'analyse de données de puces à cellules

Country Status (1)

Country Link
WO (1) WO2012125906A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10934538B2 (en) 2016-01-12 2021-03-02 Cleveland State University 3D-printed miniature biological constructs
US11262349B2 (en) 2017-10-11 2022-03-01 Cleveland State University Multiplexed immune cell assays on a micropillar/microwell chip platform
US11390836B2 (en) 2016-11-17 2022-07-19 Cleveland State University Chip platforms for microarray 3D bioprinting

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050014201A1 (en) * 2001-10-25 2005-01-20 Mordechai Deuthsch Interactive transparent individual cells biochip processor
US6977722B2 (en) * 2001-06-29 2005-12-20 Meso Scale Technologies, Llc. Assay plates, reader systems and methods for luminescence test measurements
US20090055147A1 (en) * 2004-06-25 2009-02-26 National Inst. Of Adv. Industrial Science And Tech Cell network analysis system
US20090221441A1 (en) * 2005-11-01 2009-09-03 Rensselaer Polytechnic Institute Three-dimensional cellular array chip and platform for toxicology assays
US20110007178A1 (en) * 2008-03-12 2011-01-13 Koninklijke Philips Electronics N.V. Correction of spot area in measuring brightness of sample in biosensing device

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6977722B2 (en) * 2001-06-29 2005-12-20 Meso Scale Technologies, Llc. Assay plates, reader systems and methods for luminescence test measurements
US20050014201A1 (en) * 2001-10-25 2005-01-20 Mordechai Deuthsch Interactive transparent individual cells biochip processor
US20090055147A1 (en) * 2004-06-25 2009-02-26 National Inst. Of Adv. Industrial Science And Tech Cell network analysis system
US20090221441A1 (en) * 2005-11-01 2009-09-03 Rensselaer Polytechnic Institute Three-dimensional cellular array chip and platform for toxicology assays
US20110007178A1 (en) * 2008-03-12 2011-01-13 Koninklijke Philips Electronics N.V. Correction of spot area in measuring brightness of sample in biosensing device

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10934538B2 (en) 2016-01-12 2021-03-02 Cleveland State University 3D-printed miniature biological constructs
US11390836B2 (en) 2016-11-17 2022-07-19 Cleveland State University Chip platforms for microarray 3D bioprinting
US11262349B2 (en) 2017-10-11 2022-03-01 Cleveland State University Multiplexed immune cell assays on a micropillar/microwell chip platform

Similar Documents

Publication Publication Date Title
Shahin et al. A machine vision system for grading lentils
CN108139320B (zh) 用于对试样中的干扰物进行分类的基于模型的方法和装置
CA2825727C (fr) Procede d'analyse et dispositif de lecture pour micropuce
US8873815B2 (en) System and apparatus for the remote analysis of chemical compound microarrays
US7933430B2 (en) Method and apparatus for spectral mixture resolution
WO2012125906A1 (fr) Appareil et procédé pour l'analyse de données de puces à cellules
BE1025903B1 (nl) Overspraakcorrectie bij multiplexing-analyse van biologisch monster
EP1865321A1 (fr) Procédé d'affichage d'informations de détermination de substance biologiquement pertinente
Kirkegaard et al. Nuclear magnetic resonance metabolomic profiling of Day 3 and 5 embryo culture medium does not predict pregnancy outcome in good prognosis patients: a prospective cohort study on single transferred embryos
Coma et al. Process validation and screen reproducibility in high-throughput screening
CN113203870A (zh) 用于农药残留检测仪的检测方法及农药残留检测仪
CN103347995A (zh) 核酸检查装置
Safdar et al. Reviving grain quality in wheat through non‐destructive phenotyping techniques like hyperspectral imaging
EP2824444B1 (fr) Procédé de détermination, dispositif de détermination, système de détermination et programme
EP1190762A2 (fr) Méthode pour l'affichage du résultat d'une expérience d'hybridation à l'aide d'une "bio-puce"
Mizuno et al. Development of visual egg inspection system for poultry farmer using CNN with deep learning
WO2012010691A1 (fr) Système et procédé d'analyse automatique de données de dosage de cellules biologiques
US20220373536A1 (en) High-content analysis method
CN115335502A (zh) 数据获取装置、数据获取方法和生物样本观察系统
JP6696172B2 (ja) 分析装置、分析方法、および、分析システム
JP7373659B2 (ja) 自動診断分析システムにおいて検体の特性評価中に患者情報を保護するための方法および装置
JP2014023442A (ja) 遺伝子検査装置、遺伝子検査用試薬、および遺伝子検査方法
Petter et al. Support provided by EPPO and Euphresco on phytoplasma diagnostics: recent activities including the integration of Q-bank
Lin et al. An automatic inspection and control method for a biological reagent production system with image processing techniques
CN1795451A (zh) 用于体外和体内遗传毒性试验的全自动机器人辅助图像分析系统和方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 12758258

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 12758258

Country of ref document: EP

Kind code of ref document: A1