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Standardization of tissue specimen preservation by ultrasound and temperature control

A technology of tissue samples and ultrasound, applied in the preservation of human or animal bodies, biochemical instruments, electric/wave energy processing enzymes, etc., can solve problems such as fixation, inconsistent test results, and cross-linking damage of biomolecules

Inactive Publication Date: 2012-11-14
佛山百奥快可医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Formalin-fixed tissue samples have the following 2 main problems: (1) biomolecules are severely damaged by extensive cross-linking; (2) due to various fixation times, the level of cross-linking also presents different samples
In addition to the large molecular changes, ambient temperature-induced protracted irradiation of the formalin edge occurs continuously, leading to excessive fixation while large specimens have incomplete fixation in the center
These irregular fixations often lead to discrepant results in immunohistochemistry and possibly many other molecular tests

Method used

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  • Standardization of tissue specimen preservation by ultrasound and temperature control
  • Standardization of tissue specimen preservation by ultrasound and temperature control
  • Standardization of tissue specimen preservation by ultrasound and temperature control

Examples

Experimental program
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Effect test

example 1

[0106] We treated lysozyme and BSA with NBF for 60 min at different temperatures, and immediately put the cross-linked product on an SDS gel for separation. The level of cross-linking for lysozyme and BSA gradually increased between 0-40 degrees Celsius. SDS electrophoresis showed that there was a significant increase in the size of oligomers (lysozyme) or in gelatinized fluidity (BSA) at 50 °C and reached a maximum around 60 °C. This finding predicts that when lysozyme is treated with NBF at temperatures above 60 °C, a large number of aggregates start to form. This aggregation is in highly intermolecularly cross-linked lysozyme molecules and may not be resolved by SDS electrophoresis. No aggregation was observed by BSA in NBF at high temperature, perhaps because the crosslinks in BSA are always intramolecular.

[0107] As for lysozyme, the speed of cross-linking increases gradually when the temperature is increased from 0-20°C to 20-40°C, and then suddenly increases when th...

example 2

[0109] Significant cross-linking occurs after 10 minutes of incubation at 50°C. In other experiments, we tested cross-link formers for various proteins, incubating in formalin at 4°C, at room temperature, and at 50°C for 10 minutes versus overnight in formalin at room temperature. Compare. As shown in Table 5, the speed of cross-linking at 4 °C was the slowest of all proteins tested. Then, there was a larger increase in cross-linking levels after 10 minutes of incubation at 50°C, almost comparable to overnight incubation at room temperature. For all proteins except BSA, protein precipitates formed after incubation for 10 min at 50 °C and overnight at room temperature. This experiment shows that, for most proteins, a 10-minute incubation in formalin at 50°C can form significant cross-linked morphology. Similar results were also obtained after 5 min incubation (material not shown).

example 3

[0111] Tissue samples 1–4 mm thick are placed in tissue storage boxes and then submerged in a cross-linked fixative (eg, formalin, 4°C) until sonicated. Ultrasonic waves are applied to increase the temperature of the fixative alone or in combination with other heating devices to 50-80°C. This temperature needs to be maintained for 5-20 minutes.

[0112] This tissue sample is then placed in a dehydrating agent, eg, 100% ethanol. The temperature of the dehydrating agent needs to be raised to 50-75 degrees Celsius by ultrasonic alone or in combination with other heating devices. This temperature needs to be maintained by ultrasound alone or in combination with other heating devices for 5-30 minutes. This step is repeated at least once.

[0113] This tissue sample is then placed in a cleaning agent, eg, xylene or a xylene substitute. The temperature of the cleaning agent needs to be raised to 50-75 degrees Celsius by ultrasound alone or in combination with other heating device...

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Abstract

A tissue sample preservation process and device for improving and standardizing tissue preservation procedure, including placing tissue specimens in a cold fixative, performing fixative penetration at a refrigerated temperature, and accelerating fixative penetration by ultrasound Also disclosed is the application of ultrasound and temperature control in the dehydration, clearing, and impregnation steps.

Description

technical field [0001] The invention relates to the field of tissue samples, more specifically, the invention relates to a standardized tissue sample protection through ultrasonic and temperature control. Background technique [0002] 1. Overview of the Field of Tissue Protection [0003] Ischemic lamination begins immediately after tissue samples are removed from the patient. Ischemic cascades lead to the degradation of many vulnerable cellular molecules such as mRNA (messenger ribonucleic acid), dephosphorylation of proteins in tissue samples. Thus, the longer the tissue sample is ischemic, the more pre-analytical changes in the patient's tissue sample will occur. This alteration often hinders correct judgment and diagnostic and predictive reporting of sensitive molecules [Liotta 2000, Emmert-Buck 2000, Compton 2007, Hewitt 2008, Espina 2008]. Rapid freezing, which prevents molecular alterations by storing tissue samples in a deep freeze, is a standard method of preser...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02C12N13/00C12M1/42C12M1/38
CPCA01N1/0284A01N1/0278
Inventor 朱伟星
Owner 佛山百奥快可医疗科技有限公司
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