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PAXgene Tissue FIX Container (50 ml)

用于固定并稳定组织样本
  • 保持组织形态并稳定核酸
  • 组织固定化和石蜡包埋,无需福尔马林
  • 改善了从固定组织中获得的分子生物学结果
  • 可保存组织,待以后处理
  • 从同一样本中纯化RNA、miRNA、DNA和/或蛋白质
PAXgene Tissue FIX Container (50 ml)可固定并稳定组织样本(该产品具有4个标准组织保存仓,最多可处理4个样本,每个样本体积最大可达4 x 15 x 15 mm;也可处理单个样本,其最大体积为20 x 20 x 20 mm)。PAXgene Tissue FIX Containers (50 ml)为预装有50 ml PAXgene Tissue FIX溶液的单腔容器。PAXgene Tissue FIX可快速浸透并固定组织,保存组织形态。其固定效果与福尔马林固定相当,但不会损伤核酸,无核酸交联和降解。固定后,去除PAXgene Tissue FIX,并在同一容器中加入PAXgene Tissue STABILIZER。样本中的核酸、蛋白质和样本的组织形态可在室温下保持长达7天,在2–8°C、–20°C或–80°C可保存更长时间。稳定的样本可包埋在石蜡中用于组织学研究。PAXgene Tissue Kits配合补充实验方案可从上述样本中,高效进行后续的RNA、miRNA、DNA和/或蛋白质纯化。

PAXgene Tissue FIX Container (50 ml)仅可与PAXgene Tissue STABILIZER配合使用。
Cat No./ID: 765312
PAXgene Tissue FIX Container (50 ml)
For fixation and stabilization of tissue specimen: 10 prefilled Reagent Containers containing 50 ml of PAXgene Tissue FIX

仅限于科研应用,不适用于诊断,不为疾病的诊断、预防或治疗提供参考信息。


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保存组织形态。
组织在PAXgene Tissue Fix Container (50 ml)中固定并稳定;用PAXgene Tissue固定4 μm的石蜡包埋组织切片(PFPE),后用苏木素和伊红(HE)染色。[A] 大鼠肝脏,[B]大鼠肾脏,[C]大鼠肠道。整体图像为40倍放大,细节图为400倍放大。
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Yield of high-quality, high molecular weight DNA from 12 different PAXgene Tissue-fixed (PF) tissue types, processed on the QIAcube.

Twelve different PAXgene Tissue-fixed (PF) rat tissue types, including fibrous or fatty, DNA-rich and DNA-poor tissues, processed with the PAXgene Tissue DNA Kit on the QIAcube in one single run. DNA yield per 10 mg tissue.

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Absorbance ratio of high-quality, high molecular weight DNA from 12 different PAXgene Tissue-fixed, paraffin-embedded (PF) tissue types, processed on the QIAcube.

Twelve different PAXgene Tissue-fixed (PF) rat tissue types, including fibrous or fatty, DNA-rich and DNA-poor tissue, processed with the PAXgene Tissue DNA Kit on the QIAcube in a single run. Ratio of absorbance at 260 and 280 nm.

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Comparison of manual and automated procedure: DNA yield from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue.

DNA yield from 3 sections (thickness 10 µm) each of 8 different PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicate with the PAXgene Tissue DNA Kit on the QIAcube or manually.

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Comparison of manual and automated procedure: Absorbance ratio from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue.

Ratio of absorbance at 260 and 280 nm from 3 sections (thickness 10 µm) each of 8 different PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicate with the PAXgene Tissue DNA Kit on the QIAcube or manually.

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High-quality DNA from PFPE tissue with preserved morphology.

[A] Hematoxylin and eosin (H&E) staining of PAXgene Tissue-fixed, paraffin-embedded (PFPE) human colorectal cancer tissue and [B] DNA on agarose gel electrophoresis using 0.8% TBE buffered gels with 200 ng genomic DNA isolated in triplicate from 5 cases (#1–5) of human PFPE colorectral cancer. M: markers.

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DNA without chemical modification can be used for demanding downstream applications.

Multiplex and long-range PCR of DNA from PAXgene Tissue-fixed, paraffin-embedded (PFPE) human colorectal cancer tissue (modified according to Viertler et al., (2012) A new technology for stabilization of biomolecules in tissues for combined histological and molecular analyses. J. MOl. Diagn. 14, 458). [A] Multiplex PCR of 8 different genomic DNA fragments ranging from 22 to 955 bp. [B] Long-range PCR of a 5 kb genomic DNA fragment.

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Protein extracted from PFPE tissue is suitable for two-dimentional gel electrophoresis.

Non-malignant human duodenum tissue was divided into 3 samples and either flash-frozen in liquid nitrogen (cryo), or prepared as PFPE or FFPE tissue. Proteins from cryo and PFPE tissue were extracted in 2D buffer (30 mM Tris-HCl, pH 8.8, 7 M urea, 2 M thiourea, 4% CHAPS, 75 mM DTT) supplemented with protease inhibitor. Proteins from FFPE tissue were extracted in EXB Plus buffer supplemented with protease inhibitor, precipitated with acetone and resuspended in 2D buffer (as described in Guendisch et al. 2013). Samples (150 µg) were separated by 2-D PAGE. Data kindly provided by Karl-Friedrich Becker, Technical University of Munich, Germany.

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Agarose gel electrophoresis of high-quality, high molecular weight DNA from 12 different PAXgene Tissue-fixed (PF) tissue types, processed on the QIAcube.

Twelve different PAXgene Tissue-fixed (PF) rat tissue types, including fibrous or fatty, DNA-rich and DNA-poor tissue, processed with the PAXgene Tissue DNA Kit on the QIAcube in one single run. DNA from each tissue type (300 ng) on agarose gel electrophoresis using 1% TBE buffered gels; M: markers.

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Comparison of manual and automated procedure: Agarose gel electrophoresis from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue.

Agarose gel electrophoresis from 3 sections (thickness 10 µm) each of 8 different PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicate with the PAXgene Tissue DNA Kit on the QIAcube and manually. DNA from each replicate and tissue type (200 ng) on agarose gel electrophoresis using 1% TBE buffered gels; M: markers.

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Detection of nondegraded, immunoreactive phosphoproteins from human clinical PFPE tissue.

Non-malignant human uterus, breast, prostate and bladder tissue specimens were divided into 3 samples and either flash-frozen in liquid nitrogen (cryo), PAXgene Tissue-fixed, paraffin-embedded (PFPE) or formalin-fixed, paraffin-embedded (FFPE). Proteins from cryo, PFPE and FFPE tissues were extracted using the extraction buffer EXB Plus (Qproteome FFPE Tissue Kit; described in Ergin et al. 2010, Guendisch et al. 2013 and PAXgene Tissue supplementary protocols). SDS-PAGE and Western blot analysis were performed with 15 µg protein and the indicated antibodies. Data kindly provided by Karl-Friedrich Becker, Technical University of Munich, Germany.

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High concordance of miRNA expression between total RNA isolated from PFPE and fresh-frozen tissue.

RNA, including miRNA, was purified from mirrored human breast cancer tissue fresh frozen in liquid nitrogen using the QIAGEN miRNeasy Kit, or from sections of PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue using the PAXgene Tissue RNA/miRNA Kit. Shown is a scatterplot of CT values from different single miRNA-specific RT-qPCR assays using the QIAGEN miScript System: miR9, -10a, -10b, -29a, -103, -125b, -143, -145, -192 and miScript PCR controls RNUA1, RNU5A, RNU6B, SNORD25, SCARNA19, SNORA73A; R2: coefficient of determination.

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RNA purified without chemical modification from PFPE tissue using the PAXgene Tissue RNA/miRNA Kit.

SYBR Green real-time RT-qPCR was performed with 10 ng RNA from cryopreserved, formalin-fixed, paraffin-embedded (PPFE) or PAXgene Tissue-fixed, paraffin-embedded (PFPE) rat tissue (modified according to Groelz et al. 2013). Depicted are the average delta-CT values (delta-CT = CT[FFPE] – CT[cryo] or delta-CT = CT[PFPE] – CT[cryo]) from 6 different assays with amplicons ranging from 109 to 465 bp.

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Immunohistochemistry (IHC) staining with the PAXgene Tissue System gives comparable results to formalin-fixed tissue.

Human palatine tonsil tissue was fixed in PAXgene Tissue FIX or with neutral-buffered formalin and embedded in paraffin. Primary antibodies to the indicated antigens were linked to a streptavidin-peroxidase conjugate by a biotinylated secondary antibody (LSAB method). Sectiosn were counterstained with hematoxylin. PFPE: PAXgene Tissue-fixed, paraffin-embedded. FFPE: formalin-fixed, paraffin-embedded.

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H&E staining with the PAXgene Tissue System gives results comparable to formalin-fixed tissue.

Human tissue samples were divided into 2 sub-samples. One sub-sample was fixed with PAXgene Tissue FIX and the other was fixed with neutral-buffered formalin. The fixed tissues were embedded in paraffin, sectioned and stained with hematoxylin and eosin. PFPE: PAXgene Tissue-fixed, paraffin-embedded. FFPE: formalin-fixed, paraffin-embedded.

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The PAXgene Tissue FIX Container (50 ml) and PAXgene Tissue STABILIZER workflow.

Single-chamber container for fixation and stabilization of a single, larger or multiple, smaller tissue samples.

Performance
这种固定和稳定方法在保持组织形态和核酸完整性的同时,不会产生像在福尔马林固定中产生的交联和降解现象(参见"Fixation and preservation of tissue morphology")。

固定的组织储存在PAXgene Tissue STABILIZER中时,其中的核酸、蛋白质和样本的组织形态可在室温下保持长达7天(15–25°C),或在2–8°C保存长达4周。组织样本可在PAXgene Tissue STABILIZER中,在–20°C(–15°C至–30°C)或–80°C(–65°C至–90°C)保存更长时间,而不会对组织形态或核酸完整性有不良影响。

PAXgene Tissue FIX和PAXgene Tissue STABILIZER的固定和储存条件的参数为通过动物组织测定。
Principle
目前传统组织学所使用的固定方法对分子分析的应用带来诸多限制。含有福尔马林的固定剂与生物分子交联,修饰核酸和蛋白质。在组织固定、储存和处理过程中,交联导致核酸降解。因为无法完全去除交联,其引起的化学修饰将抑制敏感的下游应用,如定量PCR或RT-PCR。为了实现对同一样本进行分子病理检测和传统病理检测,需要一种方法在固定分子的同时保存组织形态。

PreAnalytiX研制了PAXgene Tissue System以满足这些需求。该体系包括组织固定试剂(PAXgene Tissue FIX)、稳定试剂(PAXgene Tissue STABILIZER)、预装有试剂、用于组织采集、固定、储存及运输的装置、以及用于纯化RNA、DNA或包括miRNA在内的总RNA的试剂盒。此外,在www.preanalytix.com或本页面的资源选项卡下,提供纯化蛋白质的补充实验方案和其他应用信息。

容器中预装有PAXgene Tissue Reagents,配合PAXgene Tissue Kits提供了用于组织的采集、固定和稳定,以及纯化高品质核酸用于分子研究的完整解决方案。
Procedure
PAXgene Tissue FIX Containers (50 ml)为预装有50 ml PAXgene Tissue FIX溶液的单腔容器。PAXgene Tissue FIX Container (50 ml)可容纳4个标准组织保存仓(需另购),最多可处理4个样本,每个样本体积最大可达4 x 15 x 15 mm。PAXgene Tissue FIX Containers (50 ml)也可处理较大样本,其最大体积为20 x 20 x 20 mm。

PAXgene Tissue FIX可快速浸透并固定组织。固定2至48小时后(取决于组织大小),去除PAXgene Tissue FIX,并在同一容器中加入PAXgene Tissue STABILIZER。稳定的样本可包埋在石蜡中用于组织学研究。核酸和蛋白质纯化可在石蜡包埋之前后之后进行。请参见PAXgene Tissue Kit用户手册,了解DNA、RNA或miRNA的纯化信息。此外,在www.preanalytix.com或本页面的资源选项卡下,提供纯化蛋白质的补充实验方案和其他应用信息。
Applications
PAXgene Tissue试剂固定、石蜡包埋的组织切片(PFPE)可用于形态学研究或核酸或蛋白质的纯化。从PAXgene Tissue试剂固定并稳定的组织样本中纯化RNA、包括miRNA在内的总RNA或DNA时,需要使用相应的PAXgene Tissue Kit。纯化蛋白质时需要使用Qproteome FFPE Kit。

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产品介绍与指南 (6)
For isolation and purification of genomic DNA from tissue samples stabilized using the PAXgene Tissue System 

 


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For isolation and purification of total RNA, including miRNA, from tissue samples fixed and stabilized using the PAXgene Tissue System

 


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For extraction of full-length proteins from PFPE tissue using the Qproteome FFPE Tissue Kit

 


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For isolation and purification of intracellular RNA from tissue samples stabilized using the PAXgene Tissue System

 


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Moving toward excellence and standardization in tissue collection and fixation

 


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常见问题 (34)
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学术海报 (15)
Long et al., ADAPT 2012

 


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Hesse et al., AACR-NCI-EORTC 2011

 


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Groelz et al., AACR 2010

 


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Groelz et al., AMP 2010 

 


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Groelz et al., AMP 2008 

 


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Groelz et al., AMP 2009 

 


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Groelz et al., ISBER 2012

 


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Groelz et al., BRN Symposium 2012 

 


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Groelz et al., ECP 2012

 


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Groelz et al., AMP 2009

 


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Groelz et al., 3rd Annual Oncology Biomarkers 2011

 


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Groelz et al., GTEx Meeting 2014

 


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Groelz et al., AMP 2014

 


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Groelz et al., BRN Symposium 2011 

 


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Groelz et al., ECP 2014

 


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试剂盒操作手册 (2)
For collection, fixation, and stabilization of multiple small tissue samples or a single large tissue sample

 


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For stabilization of tissue specimens fixed in PAXgene Tissue FIX Container

 


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补充实验方案 (16)

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参考文献
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