# US 8,541,244 B2 · In situ heat induced antigen recovery and staining apparatus and method > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,541,244 B2 - Document ID: US8541244B2 - Status: Granted - Kind: B2 - Application number: 13/117,971 - Filed: 2011-05-27 - Published: 2011-09-22 - Granted: 2013-09-24 - Earliest claimed: 2005-05-24 - Assignees: Lee H. Angros - Applicants: Lee Angros; Thomas Lee Byers - Inventors: Lee Angros; Thomas Lee Byers - Web page: https://patentgrep.com/patent/US8541244B2 ## CPC - G01N 1/30: Staining; Impregnating {; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis} - A61F 7/12: Devices for heating or cooling internal body cavities - B01L 7/00: Heating or cooling apparatus (autoclaves B01J3/04); Heat insulating devices - G01N 1/28: Preparing specimens for investigation {including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q}(mounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 {; laboratory gas handling apparatus B01L5/00}) - G01N 1/312: {for samples mounted on planar substrates} - G01N 1/44: Sample treatment involving radiation, e.g. heat - G01N 35/00029: {provided with flat sample substrates, e.g. slides (G01N35/028 takes precedence)} - G01N 35/026: {having blocks or racks of reaction cells or cuvettes} - A61F 7/0085: {Devices for generating hot or cold treatment fluids} - A61F 2007/0063: {for cooling} - A61F 2007/126: {for invasive application, e.g. for introducing into blood vessels} - B01L 7/52: {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples} - B01L 2200/16: Reagents, handling or storing thereof - B01L 2300/0822: Slides - B01L 2300/14: Means for pressure control - B01L 2400/0478: pistons - G01N 2035/00138: {Slides} - G01N 2035/0412: {Block or rack elements with a single row of samples} - Y10T 436/25: including sample preparation - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - A61F: FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS (dental prosthetics A61C) - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION ## Abstract Contemplated herein is an automated microscope slide antigen recovery and staining apparatus and method that features a plurality of individually operable miniaturized pressurizable reaction compartments for individually and independently processing a plurality of individual microscope slides. The apparatus preferably features independently movable slide support elements each having an individually heatable heating plate. Each slide support element preferably supports a single microscope slide. Each microscope slide can be enclosed within an individual pressurizable reaction compartment. Pressures exceeding 1 atm or below 1 atm can be created and maintained in the reaction compartment prior to, during or after heating of the slide begins. Because of the ability to pressurize and regulate pressure within the reaction compartment, and to individually heat each slide, each slide and a liquid solution or reagent thereon can be heated to temperatures that could not be obtained without the enclosed pressurized environment of the reaction compartment. A reagent dispensing strip having a plurality of reconfigurable reagent modules may also be used. ## Claims ### Claim 1 A method of treating a biological specimen, comprising: disposing a microscope slide having the biological specimen thereon onto a slide support element of an automated apparatus, the apparatus comprising: a plurality of reaction compartments, each having an inner space, and a plurality of slide support elements, wherein each slide support element is sized to support a single microscope slide thereon, and wherein each slide support element is associated with one of the reaction compartments; positioning the biological specimen adjacent the inner space of the reaction compartment; causing the inner space of the reaction compartment to be sealed with a pressure-tight seal to form a sealed inner space containing the biological specimen; and pressurizing the sealed inner space of the reaction compartment to maintain an internal pressure in a range of 1 psig to 350 psig by heating a reagent or solution within the sealed inner space to a temperature in a range of 100° C. to 350° C. ### Claim 2 (depends on 1) The method of claim 1 wherein the sealed inner space is also pressurized by introducing a pressurized gas into the sealed inner space. ### Claim 3 (depends on 1) The method of claim 1 wherein in the step of pressurizing the sealed inner space, the internal pressure in the sealed inner space is controlled by a microprocessor. ### Claim 4 (depends on 1) The method of claim 1 wherein the plurality of reaction compartments are contained within a chamber and wherein each slide support element associated with the reaction compartment is independently movable for independently moving each microscope slide thereon to positions for placement of the microscope slide on the slide support element or for removal therefrom. ### Claim 5 (depends on 1) The method of claim 1 wherein at least a portion of the reaction compartment has a tubular shape, and at least a portion of the slide support element corresponding thereto has a tubular shape. ### Claim 6 (depends on 1) The method of claim 1 wherein the reaction compartment is sized to contain a single microscope slide. ### Claim 7 (depends on 1) The method of claim 1 comprising moving the reaction compartment to position the biological specimen adjacent the inner space of the reaction compartment. ### Claim 8 (depends on 1) The method of claim 1 wherein each reaction compartment is operable independently of each other reaction compartment. ### Claim 9 (depends on 1) The method of claim 1 wherein the sealed inner space of the reaction compartment is pressurized to a pressure in a range of 10 psig to 40 psig. ### Claim 10 (depends on 1) The method of claim 1 wherein the internal pressure to which sealed inner space of the reaction compartment is pressurized is at least one of 5, 8, 10, 12, 15, 16, 19, 20, 23, 25, and 30 psig. ### Claim 11 (depends on 1) The method of claim 1 wherein the microprocessor of the apparatus is programmable. ### Claim 12 (depends on 1) The method of claim 1 wherein the apparatus comprises a plurality of processing components which are independently movable and activatable by a microprocessor. ### Claim 13 (depends on 1) The method of claim 1 wherein the apparatus comprises processing components which are selected from the group of reagent dispensers, reagent strip support devices, reagent strips, aspiration devices, reagent conduits, rinse ports, vacuum ports, drainage ports, mixing jets, air cooling ducts, liquid cooling ducts, pressure ports, waste ports, and heating devices. ### Claim 14 (depends on 1) The method of claim 1 wherein each reaction compartment is independently pressurizable. ### Claim 15 (depends on 1) The method of claim 1 wherein the apparatus comprises a plurality of independently operable reaction modules wherein each reaction module comprises one of the reaction compartments, and comprises the slide support element which is associated with said reaction compartment. ### Claim 16 (depends on 15) The method of claim 15 wherein the apparatus further comprises a microprocessor which is programmable to automatically control the plurality of independently operable reaction modules. ### Claim 17 (depends on 1) The method of claim 1 wherein the step of positioning the biological specimen comprises moving the slide support element with the microscope slide thereon to position the biological specimen adjacent the inner space of the reaction compartment. ### Claim 18 A method of treating a biological specimen, comprising: disposing a microscope slide having the biological specimen thereon onto a slide support element of an automated apparatus, the apparatus comprising: a plurality of reaction compartments, each having an inner space, and a plurality of independently movable slide support elements, wherein each slide support element is sized to support a single microscope slide thereon, and wherein each slide support element is associated with a reaction compartment; positioning the biological specimen adjacent the inner space of the reaction compartment; causing the inner space of the reaction compartment to be sealed with a pressure-tight seal to form a sealed inner space containing the biological specimen; and pressurizing the sealed inner space of the reaction compartment to maintain an internal pressure therein in a range of 1 psig to 350 psig by heating a reagent or solution within the sealed inner space to a temperature in a range of 100° C. to 350° C. ### Claim 19 (depends on 18) The method of claim 18, wherein the plurality of reaction compartments are contained within a chamber and wherein each slide support element associated with the reaction compartment is independently movable for independently moving each microscope slide thereon to positions for placement of the microscope slide on the slide support element or for removal therefrom. ### Claim 20 (depends on 18) The method of claim 18, wherein at least a portion of the reaction compartment has a tubular shape, and at least a portion of the slide support element corresponding thereto has a tubular shape. ### Claim 21 (depends on 18) The method of claim 18 comprising moving the reaction compartment to position the biological specimen adjacent the inner space of the reaction compartment. ### Claim 22 (depends on 18) The method of claim 18, wherein the apparatus comprises a plurality of processing components which are independently movable and activatable by a microprocessor. ### Claim 23 (depends on 18) The method of claim 18 wherein the step of positioning the biological specimen comprises moving the slide support element with the microscope slide thereon to position the biological specimen adjacent the inner space of the reaction compartment. ### Claim 24 A method of treating a biological specimen, comprising: disposing a microscope slide having the biological specimen thereon onto a slide support element of an automated apparatus, the apparatus comprising: a plurality of reaction compartments, each having an inner space, wherein at least a portion of each reaction compartment has a tubular shape, and a plurality of independently movable slide support elements, wherein each slide support element is sized to support a single microscope slide thereon, and wherein each slide support element is associated with a reaction compartment; positioning the biological specimen adjacent the inner space of the reaction compartment; causing the inner space of the reaction compartment to be sealed with a pressure-tight seal to form a sealed inner space containing the biological specimen; and pressurizing the sealed inner space of the reaction compartment to maintain an internal pressure therein in a range of from 1 psig to 350 psig by heating a reagent or solution within the sealed inner space to a temperature in a range of 100° C. to 350° C. ### Claim 25 (depends on 24) The method of claim 24, wherein the plurality of reaction compartments are contained within a chamber and wherein each slide support element associated with the reaction compartment is independently movable for independently moving each microscope slide thereon to positions for placement of the microscope slide on the slide support element or for removal therefrom. ### Claim 26 (depends on 24) The method of claim 24, wherein at least a portion of the slide support element associated with the reaction compartment has a tubular shape. ### Claim 27 (depends on 24) The method of claim 24 comprising moving the reaction compartment to position the biological specimen adjacent the inner space of the reaction compartment. ### Claim 28 (depends on 24) The method of claim 24, wherein the apparatus comprises a plurality of processing components which are independently movable and activatable by a microprocessor. ### Claim 29 (depends on 24) The method of claim 24 wherein the step of positioning the biological specimen comprises moving the slide support element with the microscope slide thereon to position the biological specimen adjacent the inner space of the reaction compartment. ### Claim 30 A method of treating a biological specimen, comprising: disposing a microscope slide having the biological specimen thereon onto a slide support element of an automated apparatus, the apparatus comprising: a plurality of reaction compartments, each having an inner space, and a plurality of slide support elements, wherein each slide support element is sized to support a single microscope slide thereon, and wherein each slide support element is associated with a reaction compartment; positioning the biological specimen within the inner space of the reaction compartment; causing the inner space of the reaction compartment to be sealed with a pressure-tight seal to form a sealed inner space containing the biological specimen; and pressurizing the sealed inner space of the reaction compartment to maintain an internal pressure therein in a range of 1 psig to 350 psig by heating a reagent or solution within the sealed inner space to a temperature in a range of 100° C. to 350° C. ### Claim 31 (depends on 30) The method of claim 30, wherein the plurality of reaction compartments are contained within a chamber and wherein each slide support element associated with the reaction compartment is independently movable for independently moving each microscope slide thereon to positions for placement of the microscope slide on the slide support element or for removal therefrom. ### Claim 32 (depends on 30) The method of claim 30, wherein at least a portion of the reaction compartment has a tubular shape, and at least a portion of the slide support element corresponding thereto has a tubular shape. ### Claim 33 (depends on 30) The method of claim 30 comprising moving the reaction compartment to position the biological specimen within the inner space of the reaction compartment. ### Claim 34 (depends on 30) The method of claim 30, wherein the apparatus comprises a plurality of processing components which are independently movable and activatable by a microprocessor. ### Claim 35 (depends on 30) The method of claim 30 wherein the step of positioning the biological specimen comprises moving the slide support element with the microscope slide thereon to position the biological specimen within the inner space of the reaction compartment. ### Claim 36 A method of treating a biological specimen, comprising: disposing a microscope slide having the biological specimen thereon onto a slide support element of an automated apparatus, the apparatus comprising: a plurality of reaction compartments, each having an inner space, and a plurality of independently movable slide support elements, wherein each slide support element is sized to support a single microscope slide thereon, and wherein each slide support element is associated with a reaction compartment; positioning the biological specimen within the inner space of the reaction compartment; causing the inner space of the reaction compartment to be sealed with a pressure-tight seal to form a sealed inner space containing the biological specimen; and pressurizing the sealed inner space of the reaction compartment to maintain an internal pressure therein in a range of 1 psig to 350 psig by heating a reagent or solution within the sealed inner space to a temperature in a range of 100° C. to 350° C. ### Claim 37 (depends on 36) The method of claim 36, wherein the plurality of reaction compartments are contained within a chamber and wherein each slide support element associated with the reaction compartment is independently movable for independently moving each microscope slide thereon to positions for placement of the microscope slide on the slide support element or for removal therefrom. ### Claim 38 (depends on 36) The method of claim 36, wherein at least a portion of the reaction compartment has a tubular shape, and at least a portion of the slide support element corresponding thereto has a tubular shape. ### Claim 39 (depends on 36) The method of claim 36 comprising moving the reaction compartment to position the biological specimen within the inner space of the reaction compartment. ### Claim 40 (depends on 36) The method of claim 36, wherein the apparatus comprises a plurality of processing components which are independently movable and activatable by a microprocessor. ### Claim 41 (depends on 36) The method of claim 36 wherein the step of positioning the biological specimen comprises moving the slide support element with the microscope slide thereon to position the biological specimen within the inner space of the reaction compartment. ### Claim 42 A method of treating a biological specimen, comprising: disposing a microscope slide having the biological specimen thereon onto a slide support element of an automated apparatus, the apparatus comprising: a plurality of reaction compartments, each having an inner space, wherein at least a portion of each reaction compartment has a tubular shape, and a plurality of independently movable slide support elements, wherein each slide support element is sized to support a single microscope slide thereon, and wherein each slide support element is associated with a reaction compartment; positioning the biological specimen within the inner space of the reaction compartment; causing the inner space of the reaction compartment to be sealed with a pressure-tight seal to form a sealed inner space containing the biological specimen; and pressurizing the sealed inner space of the reaction compartment to maintain an internal pressure therein in a range of from 1 psig to 350 psig by heating a reagent or solution within the sealed inner space to a temperature in a range of 100° C. to 350° C. ### Claim 43 (depends on 42) The method of claim 42, wherein the plurality of reaction compartments are contained within a chamber and wherein each slide support element associated with the reaction compartment is independently movable for independently moving each microscope slide thereon to positions for placement of the microscope slide on the slide support element or for removal therefrom. ### Claim 44 (depends on 42) The method of claim 42, wherein at least a portion of the slide support element associated with the reaction compartment has a tubular shape. ### Claim 45 (depends on 42) The method of claim 42 comprising moving the reaction compartment to position the biological specimen within the inner space of the reaction compartment. ### Claim 46 (depends on 42) The method of claim 42, wherein the apparatus comprises a plurality of processing components which are independently movable and activatable by a microprocessor. ### Claim 47 (depends on 42) The method of claim 42 wherein the step of positioning the biological specimen comprises moving the slide support element with the microscope slide thereon to position the biological specimen within the inner space of the reaction compartment. ### Claim 48 A method of treating a biological specimen, comprising: disposing a microscope slide having the biological specimen thereon onto a slide support element of an automated apparatus, the apparatus comprising: a plurality of reaction compartments, each having an inner space; and a plurality of independently movable slide support elements, each sized to support a single microscope slide, and wherein each slide support element is movable to a position within or adjacent the reaction compartment corresponding thereto and wherein each slide support element can be turned within the reaction compartment corresponding thereto; positioning the biological specimen within or adjacent the inner space of the reaction compartment; causing the inner space of the reaction compartment to be sealed with a pressure-tight seal to form a sealed inner space of the reaction compartment containing the biological specimen; and pressurizing the sealed inner space of each reaction compartment, and wherein each reaction compartment is pressurizable independently of each other reaction compartment, wherein a microprocessor is used to control the pressurization of the sealed inner space to individually pressurize and maintain the sealed inner space of the reaction compartment at a pressure level above atmospheric pressure. ### Claim 49 (depends on 48) The method of claim 48, wherein the plurality of reaction compartments are contained within a chamber and wherein each slide support element associated with the reaction compartment is independently movable for independently moving each microscope slide thereon to positions for placement of the microscope slide on the slide support element or for removal therefrom. ### Claim 50 (depends on 48) The method of claim 48, wherein at least a portion of the reaction compartment has a tubular shape, and at least a portion of the slide support element corresponding thereto has a tubular shape. ### Claim 51 (depends on 48) The method of claim 48 comprising moving the reaction compartment to position the biological specimen within or adjacent the inner space of the reaction compartment. ### Claim 52 (depends on 48) The method of claim 48, wherein the apparatus comprises a plurality of processing components which are independently movable and activatable by the microprocessor. ### Claim 53 (depends on 48) The method of claim 48, wherein the pressure level is maintained in the reaction compartment in a range of from 1 psig to 350 psig, and wherein the pressurization occurs by heating a reagent or solution within the sealed inner space to a temperature in a range of 100° C. to 350° C. ### Claim 54 (depends on 48) The method of claim 48 wherein the step of positioning the biological specimen comprises moving the slide support element with the microscope slide thereon to position the biological specimen within or adjacent the inner space of the reaction compartment.