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Two phase immiscible system for the pretreatment of embedded biological samples

一种生物样本、预处理的技术，应用在测试用样品的制备、取样、仪器等方向，能够解决升高温度、效率低、缓慢等问题

Active Publication Date: 2017-10-17
AGILENT TECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0020] A disadvantage of the method and system for removing embedding medium with heat is that it can be a slow process, as paraffin-embedded biological samples are subjected to elevated temperatures over a period of 5 minutes to 60 minutes
Another disadvantage is the low efficiency when removing the last paraffin residues in tissue sections
A further disadvantage is that the heating elements used require sufficient contact between the surface on which the biological sample is placed and the heating element

Method used

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  • Two phase immiscible system for the pretreatment of embedded biological samples
  • Two phase immiscible system for the pretreatment of embedded biological samples
  • Two phase immiscible system for the pretreatment of embedded biological samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0209] This experiment was performed to reconfirm that manually performed (by hand) two-phase deparaffinization can be compared with the Dako standard formamide HER2FISH pharmDx TM Use together. Perform the target retrieval step in the microwave using MES buffer.

[0210] -Add DI water to the container

[0211] - Add solvent on top of DI water (Histoclear ) (forming a two-phase system)

[0212] - Immerse the slide in the two-phase system

[0213] - Remove the slide from the two-phase system

[0214] -Incubate for 2 minutes

[0215] - Wash 2 x 3 min in wash buffer solution

[0216] - Target repair for 10 minutes in the microwave

[0217] -Continue to standard procedure

[0218] Conclusions: Manually performed two-phase deparaffinization with traditional formamide buffer is suitable for Dako FISH and CISH. Remnants (droplets) of Histoclear were present on slides in very high concentrations (Histoclear was not removed before the TR step).

Embodiment 2

[0220] This experiment was performed to test whether manual (by hand) 2-phase deparaffinization could be performed with HER2IQFISH PharmDx as compared to conventional xylene processing. TM buffer used together. The target restoration step is performed in a microwave oven. Note that the CISH operational guidelines used are very demanding on the organization.

[0221] -Add DI water to the container

[0222] -Add solvent (Histoclear) on top of DI water (two-phase system)

[0223] - Immerse the slide in the two-phase system

[0224] - Remove the slide from the two-phase system

[0225] -Incubate for 2 minutes

[0226] - Wash 3 x 3 min in wash buffer solution

[0227] - Target repair for 10 minutes in the microwave

[0228] -Continuation of FISH / CISH procedure

[0229] Conclusion: Using HER2IQFISH PharmDx TM Manually performed 2-phase deparaffinization of buffers is suitable for Dako FISH and CISH. Histoclear The residues (droplets) were present on the slide in very hig...

Embodiment 3

[0231] This experiment is for testing, compared with traditional xylene treatment, automation ( Figure 8-9 ) Can phase 2 deparaffinization work with HER2IQFISH PharmDx TM Use together. The object restoration step is performed by the module.

[0232] Version A:

[0233] - Add DI water to the bottom of the tank

[0234] - the solvent (Histoclear ) on top of DI water (liquid position 1), 2-phase system

[0235] -Fill the tank with DI water

[0236] - Empty tank to liquid position 1

[0237] - Incubate for 2 minutes.

[0238] - Fill tank with DI water and overflow (Histoclear run off in the overflow drain)

[0239] - empty the slot

[0240] -Fill the tank with MES buffer

[0241] -Incubate at 97°C for 10 minutes

[0242] - Rinse with DI water to 35°C (remaining Histoclear out the drain)

[0243] - Transfer slides to wash buffer

[0244] -Continue to FISH / CISH procedure

[0245] Version B:

[0246] - Add MES buffer to the bottom of the tank

[0247] - the solv...

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Abstract

The present application provides a two phase immiscible system for the pretreatment of embedded biological samples comprising placing at least one support having an embedded biological sample on its surface into a pretreatment container, adding to 5 the pretreatment container at least one reagent forming a layer, adding a carrier composition to the pretreatment container, such that reagent forming layer is formed on the top of the carrier composition, and in an amount such that the at least one reagent forming layer contacts at least a portion of the embedded biological sample. Pretreatment of the embedded biological samples can include removal of embedding medium from embedded biological, target retrieval and enzyme blocking samples before staining histochemical analysis or other processes. The system also includes an apparatus and processes of automation of the pretreatment methods.

Description

technical field [0001] The present disclosure relates to the field of processing of biological samples, in particular pretreatment of embedded biological samples, such as removal of embedding medium from embedded biological samples prior to staining, histochemical analysis or other procedures. More specifically, the present invention relates to the application of a two-phase system having an immersion tank and a surface layer in a procedure to remove embedding medium from embedded biological samples. The present invention relates to an efficient and cost-effective method and composition for removing embedding medium from embedded biological samples. Background technique [0002] Sample processing, such as in immunohistochemistry ("IHC") applications, as well as in other chemical and biological analyses, may involve at least one processing sequence or processing protocol as part of the analysis of at least one sample. Typically, such processing guidelines are established by ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30G01N1/36
CPCG01N1/30G01N1/36G01N2001/366G01N1/312
Inventor S·H·马蒂森S·尼尔森
Owner AGILENT TECH INC
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