# US 10,416,052 B2 · In situ heat induced antigen recovery and staining apparatus and method

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- Number: US 10,416,052 B2
- Document ID: US10416052B2
- Status: Granted
- Kind: B2
- Application number: 15/289,675
- Filed: 2016-10-10
- Published: 2017-01-26
- Granted: 2019-09-17
- Earliest claimed: 1999-07-08
- Applicants: Lee H. Angros
- Inventors: Lee H. Angros
- Web page: https://patentgrep.com/patent/US10416052B2

## 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}
- G01N 1/312: {for samples mounted on planar substrates}
- G01N 1/44: Sample treatment involving radiation, e.g. heat
- G01N 33/52: Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper {and including single- and multilayer analytical elements (immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72)}
- G01N 33/521: {Single-layer analytical elements}
- G01N 33/53: Immunoassay; Biospecific binding assay; Materials therefor
- G01N 33/5306: {Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding}
- G01N 35/00029: {provided with flat sample substrates, e.g. slides (G01N35/028 takes precedence)}
- G01N 35/00584: {Control arrangements for automatic analysers}
- B01L 9/52: {Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips}
- G01N 2035/00138: {Slides}
- G01N 2035/00346: {Heating or cooling arrangements}
- Y10T 436/11: Automated chemical analysis
- Y10T 436/110833: Utilizing a moving indicator strip or tape
- Y10T 436/111666: Utilizing a centrifuge or compartmented rotor
- Y10T 436/112499: with sample on test slide
- Y10T 436/114165: with step of insertion or removal from test line
- Y10T 436/114998: with treatment or replacement of aspirator element [e.g., cleaning, etc.]
- Y10T 436/119163: with aspirator of claimed structure
- Y10T 436/25: including sample preparation
- Y10T 436/2575: Volumetric liquid transfer
- 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)
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

An automated in situ heat induced antigen recovery and staining method and apparatus for treating a plurality of microscope slides. The process of heat induced antigen recovery and the process of staining the biological sample on the microscope slide are conducted in the same apparatus, wherein the microscope slides do not need to be physically removed from one apparatus to another. Each treatment step occurs within the same reaction compartment. The reaction conditions of each reaction compartment for treating a slide can preferably be controlled independently, including the individualized application of reagents to each slide and the individualized treatment of each slide.

## Claims

### Claim 1

A method of treating biological samples, comprising:
placing a first microscope slide on a first slide support element with the first microscope slide having a first biological sample disposed thereon;
placing a second microscope slide on a second slide support element with the second microscope slide having a second biological sample disposed thereon;
horizontally moving the first slide support into a treatment chamber of an antigen recovery and staining apparatus so as to position the first microscope slide and the first biological sample in the treatment chamber;
horizontally moving the second slide support into the treatment chamber of the antigen recovery and staining apparatus so as to position the second microscope slide and the second biological sample in the treatment chamber;
treating the first biological sample by applying a first reagent to the first biological sample, heating the first reagent to a desired temperature for a desired period of time, and applying additional reagents of a treatment protocol;
treating the second biological sample by applying a second reagent to the second biological sample, heating the second reagent to a desired temperature for a desired period of time, and applying additional reagents of another treatment protocol;
determining if one of the first biological sample and the second biological sample has completed the treatment protocol; and
removing the microscope slide with the biological sample that has completed the treatment protocol from the treatment chamber while the other biological sample continues to be treated in the treatment chamber,
wherein the step of removing the microscope slide with the biological sample that has completed the treatment protocol from the treatment chamber comprises the steps of:
horizontally moving the slide support out of the treatment chamber of the antigen recovery and staining apparatus so as to position the microscope slide with the biological sample that has completed the treatment protocol outside of the treatment chamber; and
removing the microscope slide with the biological sample that has completed the treatment protocol from the slide support element.

### Claim 2 (depends on 1)

The method of claim 1 further comprising the steps of:
positioning the first biological sample in a first reaction compartment and positioning the second biological sample in a second reaction compartment,
wherein the first reaction compartment is separate from the second reaction compartment.

### Claim 3 (depends on 2)

The method of claim 2 wherein the step of positioning the first biological sample in the first reaction compartment, the first microscope slide defines a portion of the first reaction compartment, and wherein the step of positioning the second biological sample in the second reaction compartment, the second microscope slide defines a portion of the second reaction compartment.

### Claim 4 (depends on 2)

The method of claim 2 wherein the step of positioning the first biological sample in the first reaction compartment, the first slide support element defines a portion of the first reaction compartment, and wherein the step of positioning the second biological sample in the second reaction compartment, the second slide support element defines a portion of the second reaction compartment.

### Claim 5 (depends on 1)

The method of claim 1 further comprising the steps of:
placing a third microscope slide on the slide support element from which the microscope slide with the biological sample that has completed the treatment protocol has been removed with the third microscope slide having a third biological sample disposed thereon; and
horizontally moving the slide support with the third microscope slide positioned thereon into a treatment chamber of an antigen recovery and staining apparatus so as to position the third microscope slide and the third biological sample in the treatment chamber while the other biological sample continues to be treated in the treatment chamber.

### Claim 6

A method of treating biological samples, comprising:
placing a first microscope slide on a first slide support element with the first microscope slide having a first biological sample disposed thereon;
horizontally moving the first slide support into a treatment chamber of an antigen recovery and staining apparatus so as to position the first microscope slide and the first biological sample in the treatment chamber;
treating the first biological sample by applying a first reagent to the first biological sample, heating the first reagent to a desired temperature for a desired period of time, and applying additional reagents of a treatment protocol;
placing a second microscope slide position on a second slide support element with the second microscope slide having a second biological sample disposed thereon; and
horizontally moving the second slide support into the treatment chamber of the antigen recovery and staining apparatus so as to position the second microscope slide and the second biological sample in the treatment chamber while the first biological sample continues to be treated in the treatment chamber.

### Claim 7 (depends on 6)

The method of claim 6 further comprising the steps of:
treating the second biological sample by applying a second reagent to the second biological sample, heating the second reagent to a desired temperature for a desired period of time, and applying additional reagents of another treatment protocol;
determining if one of the first biological sample and the second biological sample has completed the treatment protocol; and
removing the microscope slide with the biological sample that has completed the treatment protocol from the treatment chamber while the other biological sample continues to be treated in the treatment chamber,
wherein the step of removing the microscope slide with the biological sample that has completed the treatment protocol from the treatment chamber comprises the steps of:
horizontally moving the slide support out of the treatment chamber of the antigen recovery and staining apparatus so as to position the microscope slide with the biological sample that has completed the treatment protocol outside of the treatment chamber; and
removing the microscope slide with the biological sample that has completed the treatment protocol from the slide support element.

### Claim 8 (depends on 6)

The method of claim 6 further comprising the steps of:
positioning the first biological sample in a first reaction compartment and positioning the second biological sample in a second reaction compartment,
wherein the first reaction compartment is separate from the second reaction compartment.

### Claim 9 (depends on 8)

The method of claim 8 wherein the step of positioning the first biological sample in the first reaction compartment, the first microscope slide defines a portion of the first reaction compartment, and wherein the step of positioning the second biological sample in the second reaction compartment, the second microscope slide defines a portion of the second reaction compartment.

### Claim 10 (depends on 8)

The method of claim 8 wherein the step of positioning the first biological sample in the first reaction compartment, the first slide support element defines a portion of the first reaction compartment, and wherein the step of positioning the second biological sample in the second reaction compartment, the second slide support element defines a portion of the second reaction compartment.

### Claim 11

A method of treating biological samples, comprising:
placing a first microscope slide on a first slide support element with the first microscope slide having a first biological sample disposed thereon;
placing a second microscope slide on a second slide support element with the second microscope slide having a second biological sample disposed thereon;
horizontally moving the first slide support into a treatment chamber of an antigen recovery and staining apparatus so as to position the first microscope slide and the first biological sample in the treatment chamber;
horizontally moving the second slide support into the treatment chamber of the antigen recovery and staining apparatus so as to position the second microscope slide and the second biological sample in the treatment chamber;
treating the first biological sample by applying a first reagent to the first biological sample, heating the first reagent to a desired temperature for a desired period of time, and applying additional reagents of a treatment protocol;
treating the second biological sample by applying a second reagent to the second biological sample, heating the second reagent to a desired temperature for a desired period of time, and applying additional reagents of another treatment protocol;
determining if one of the first biological sample and the second biological sample has completed the treatment protocol; and
horizontally removing the microscope slide with the biological sample that has completed the treatment protocol from the treatment chamber while the other biological sample continues to be treated in the treatment chamber.

### Claim 12 (depends on 11)

The method of claim 11 further comprising the steps of:
positioning the first biological sample in a first reaction compartment and positioning the second biological sample in a second reaction compartment,
wherein the first reaction compartment is separate from the second reaction compartment.

### Claim 13 (depends on 12)

The method of claim 12 wherein the step of positioning the first biological sample in the first reaction compartment, the first microscope slide defines a portion of the first reaction compartment, and wherein the step of positioning the second biological sample in the second reaction compartment, the second microscope slide defines a portion of the second reaction compartment.

### Claim 14 (depends on 12)

The method of claim 12 wherein the step of positioning the first biological sample in the first reaction compartment, the first slide support element defines a portion of the first reaction compartment, and wherein the step of positioning the second biological sample in the second reaction compartment, the second slide support element defines a portion of the second reaction compartment.

### Claim 15

A method of treating biological samples, comprising:
placing a first microscope slide on a first slide support element with the first microscope slide having a first biological sample disposed thereon;
placing a second microscope slide on a second slide support element with the second microscope slide having a second biological sample disposed thereon;
horizontally moving the first slide support into an antigen recovery and staining apparatus;
horizontally moving the second slide support into the antigen recovery and staining apparatus;
treating the first biological sample by applying a first reagent to the first biological sample, heating the first reagent to a desired temperature for a desired period of time, and applying additional reagents of a treatment protocol;
treating the second biological sample by applying a second reagent to the second biological sample, heating the second reagent to a desired temperature for a desired period of time, and applying additional reagents of another treatment protocol;
determining if one of the first biological sample and the second biological sample has completed the treatment protocol; and
removing the microscope slide with the biological sample that has completed the treatment protocol from the antigen recovery and staining apparatus while the other biological sample continues to be treated in the antigen recovery and staining apparatus,
wherein the step of removing the microscope slide with the biological sample that has completed the treatment protocol from the antigen recovery and staining apparatus comprises the steps of:
horizontally moving the slide support out of the antigen recovery and staining apparatus so as to position the microscope slide with the biological sample that has completed the treatment protocol outside of the antigen recovery and staining apparatus; and
removing the microscope slide with the biological sample that has completed the treatment protocol from the slide support element.

### Claim 16 (depends on 15)

The method of claim 15 further comprising the steps of:
positioning the first biological sample in a first reaction compartment and positioning the second biological sample in a second reaction compartment,
wherein the first reaction compartment is separate from the second reaction compartment.

### Claim 17 (depends on 16)

The method of claim 16 wherein the step of positioning the first biological sample in the first reaction compartment, the first microscope slide defines a portion of the first reaction compartment, and wherein the step of positioning the second biological sample in the second reaction compartment, the second microscope slide defines a portion of the second reaction compartment.

### Claim 18 (depends on 16)

The method of claim 16 wherein the step of positioning the first biological sample in the first reaction compartment, the first slide support element defines a portion of the first reaction compartment, and wherein the step of positioning the second biological sample in the second reaction compartment, the second slide support element defines a portion of the second reaction compartment.

### Claim 19 (depends on 15)

The method of claim 15 further comprising the steps of:
placing a third microscope slide in a horizontal position on the slide support element from which the microscope slide with the biological sample that has completed the treatment protocol has been removed with the third microscope slide having a third biological sample disposed thereon; and
horizontally moving the slide support with the third microscope slide positioned thereon into the antigen recovery and staining apparatus so as to position the third microscope slide and the third biological sample in the antigen recovery and staining apparatus while the other biological sample continues to be treated in the antigen recovery and staining apparatus.

### Claim 20

A method of treating biological samples, comprising:
placing a first microscope slide on a first slide support element with the first microscope slide having a first biological sample disposed thereon;
horizontally moving the first slide support into an antigen recovery and staining apparatus;
treating the first biological sample by applying a first reagent to the first biological sample, heating the first reagent to a desired temperature for a desired period of time, and applying additional reagents of a treatment protocol;
placing a second microscope slide on a second slide support element with the second microscope slide having a second biological sample disposed thereon; and
horizontally moving the second slide support into the antigen recovery and staining apparatus so as to position the second microscope slide and the second biological sample in the antigen recovery and staining apparatus while the first biological sample continues to be treated in the antigen recovery and staining apparatus.

### Claim 21 (depends on 20)

The method of claim 20 further comprising the steps of:
treating the second biological sample by applying a second reagent to the second biological sample, heating the second reagent to a desired temperature for a desired period of time, and applying additional reagents of another treatment protocol;
determining if one of the first biological sample and the second biological sample has completed the treatment protocol; and
removing the microscope slide with the biological sample that has completed the treatment protocol from the treatment chamber while the other biological sample continues to be treated in the treatment chamber,
wherein the step of removing the microscope slide with the biological sample that has completed the treatment protocol from the antigen recovery and staining apparatus comprises the steps of:
horizontally moving the slide support out of the antigen recovery and staining apparatus so as to position the microscope slide with the biological sample that has completed the treatment protocol outside of the antigen recovery and staining apparatus; and
removing the microscope slide with the biological sample that has completed the treatment protocol from the slide support element.

### Claim 22 (depends on 20)

The method of claim 20 further comprising the steps of:
positioning the first biological sample in a first reaction compartment and positioning the second biological sample in a second reaction compartment,
wherein the first reaction compartment is separate from the second reaction compartment.

### Claim 23 (depends on 22)

The method of claim 22 wherein the step of positioning the first biological sample in the first reaction compartment, the first microscope slide defines a portion of the first reaction compartment, and wherein the step of positioning the second biological sample in the second reaction compartment, the second microscope slide defines a portion of the second reaction compartment.

### Claim 24 (depends on 22)

The method of claim 22 wherein the step of positioning the first biological sample in the first reaction compartment, the first slide support element defines a portion of the first reaction compartment, and wherein the step of positioning the second biological sample in the second reaction compartment, the second slide support element defines a portion of the second reaction compartment.

### Claim 25

A method of treating biological samples, comprising:
placing a first microscope slide on a first slide support element with the first microscope slide having a first biological sample disposed thereon;
placing a second microscope slide on a second slide support element with the second microscope slide having a second biological sample disposed thereon;
horizontally moving the first slide support into an antigen recovery and staining apparatus;
horizontally moving the second slide support into the antigen recovery and staining apparatus;
treating the first biological sample by applying a first regent to the first biological sample, heating the first reagent to a desired temperature for a desired period of time, and applying additional reagents of a treatment protocol;
treating the second biological sample by applying a second reagent to the second biological sample, heating the second reagent to a desired temperature for a desired period of time, and applying additional reagents of another treatment protocol;
determining if one of the first biological sample and the second biological sample has completed the treatment protocol; and
horizontally removing the microscope slide with the biological sample that has completed the treatment protocol from the antigen recovery and staining apparatus while the other biological sample continues to be treated in the antigen recovery and staining apparatus.

### Claim 26 (depends on 25)

The method of claim 25 further comprising the steps of:
positioning the first biological sample in a first reaction compartment and positioning the second biological sample in a second reaction compartment,
wherein the first reaction compartment is separate from the second reaction compartment.

### Claim 27 (depends on 26)

The method of claim 26 wherein the step of positioning the first biological sample in the first reaction compartment, the first microscope slide defines a portion of the first reaction compartment, and wherein the step of positioning the second biological sample in the second reaction compartment, the second microscope slide defines a portion of the second reaction compartment.

### Claim 28 (depends on 26)

The method of claim 26 wherein the step of positioning the first biological sample in the first reaction compartment, the first slide support element defines a portion of the first reaction compartment, and wherein the step of positioning the second biological sample in the second reaction compartment, the second slide support element defines a portion of the second reaction compartment.