# US 12,216,089 B2 · Processing system for processing specimens using acoustic energy

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- Number: US 12,216,089 B2
- Document ID: US12216089B2
- Status: Granted
- Kind: B2
- Application number: 18/107,951
- Filed: 2023-02-09
- Published: 2023-06-15
- Granted: 2025-02-04
- Earliest claimed: 2010-03-04
- Assignees: Ventana Medical Systems, Inc.
- Applicants: Ventana Medical Systems, Inc.
- Inventors: David Chafin; Michael Otter; Abbey Pierson; Jefferson Curtis Taft
- Web page: https://patentgrep.com/patent/US12216089B2

## CPC

- G01N 29/07: by measuring propagation velocity or propagation time of acoustic waves
- 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 29/024: by measuring propagation velocity or propagation time of acoustic waves
- G01N 29/4427: {with stored values, e.g. threshold values}
- G01N 33/4833: {of solid biological material, e.g. tissue samples, cell cultures (tissue in vivo A61B5/00; cell suspensions G01N33/48735)}
- G01N 2291/02466: Biological material, e.g. blood
- G01N 2291/02475: Tissue characterisation
- 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)

## Abstract

A method for fixing a biological sample includes delivering energy through a biological sample that has been removed from a subject, while fixing the biological sample. A change in speed of the energy traveling through the biological sample is evaluated to monitor the progress of the fixation. A system for performing the method can include a transmitter that outputs the energy and a receiver configured to detect the transmitted energy. A computing device can evaluate the speed of the energy based on signals from the receiver.

## Claims

### Claim 1

A method of processing a tissue sample comprising:
(i) immersing the tissue sample in a fixative solution having a first temperature ranging from 0° C. to 10° C. for a first time period sufficient to diffuse the fixative solution into an interior region of the tissue sample without substantially forming cross-links within the tissue sample; and
(ii) continuing immersion of the tissue sample in the fixative solution for a second period of time sufficient to cause cross-linking within the tissue sample,
wherein the tissue sample is not exposed to ultrasonic or electromagnetic irradiation during either (i) the immersion of the tissue sample in the fixative solution, or (ii) the continuing immersion of the tissue sample in the fixative solution.

### Claim 2 (depends on 1)

The method of claim 1, wherein the first time period is about 2 hours.

### Claim 3 (depends on 1)

The method of claim 1, wherein the first temperature is about 4° C.

### Claim 4 (depends on 1)

The method of claim 1, wherein the immersion in the fixative solution for the second period of time is continued by submerging the tissue sample in a warm fixative bath.

### Claim 5 (depends on 4)

The method of claim 4, wherein the warm fixative bath has a temperature of about 45° C.

### Claim 6 (depends on 5)

The method of claim 5, wherein the second time period is about 2 hours.

### Claim 7 (depends on 1)

The method of claim 1, wherein the immersion in the fixative solution for the second time period is continued by heating the fixative solution from the first temperature to a second temperature.

### Claim 8 (depends on 1)

The method of claim 1, wherein the fixative solution is 10% NBF.

### Claim 9 (depends on 1)

The method of claim 1, wherein the second time period is about 2 hours.

### Claim 10 (depends on 1)

The method of claim 1, wherein the second time period is about 4 hours.

### Claim 11 (depends on 1)

The method of claim 1, wherein the second time period is about 6 hours.

### Claim 12 (depends on 1)

The method of claim 1, where the first time period is sufficient to diffuse the fixative solution throughout substantially all of the tissue sample.

### Claim 13 (depends on 1)

The method of claim 1, further comprising monitoring the diffusion of the fixative solution through the tissue sample.

### Claim 14 (depends on 1)

The method of claim 1, wherein the monitoring comprises evaluating a change in a time of flight of acoustic waves that travel through the tissue sample while it is immersed within the fixative solution at the first temperature.

### Claim 15 (depends on 1)

The method of claim 1, further comprising monitoring the cross-linking of the tissue sample.