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PAXgene Tissue FIX Container (50 ml)

For collection, fixation and stabilization of multiple small or a single large tissue sample
  • Formalin-free preservation of morphology and biomolecules
  • Improved molecular results from fixed tissues
  • Tissue can be stored and archived for later processing
  • RNA, miRNA, DNA, and/or proteins from one sample

 

The PAXgene Tissue FIX Container is intended for the collection, fixation and stabilization of tissue specimens. The container is part of the PAXgene Tissue System and must be used with the PAXgene Tissue STABILIZER Concentrate. The PAXgene Tissue System additionally includes the PAXgene RNA/miRNA Kit for isolation of total RNA, including miRNA, and PAXgene Tissue DNA Kit for DNA. Supplementary protocols are available for other applications, including purification of proteins.
Cat No./ID: 765312
PAXgene Tissue FIX Container (50 ml)
€129.00
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For fixation and stabilization of tissue specimen: 10 prefilled Reagent Containers containing 50 ml of PAXgene Tissue FIX
For Research Use Only. Not for use in diagnostics procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease.
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Preservation of tissue morphology.
Tissue was fixed and stabilized with the PAXgene Tissue FIX Container (50 ml); 4 µm sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue were stained with hematoxylin and eosin (HE). [A] Rat liver, [B] rat kidney, [C] rat intestine. Overview at 40x, details at 400x magnification.
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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
PAXgene Tissue FIX Containers are 50 ml containers prefilled with PAXgene Tissue FIX, a reagent that rapidly penetrates and fixes tissues, preserving tissue morphology and biomolecules. The container accommodates up to 4 standard tissue cassettes or a single, large tissue specimen. Fixation with PAXgene Tissue FIX is comparable to formalin fixation, but avoids destructive nucleic acid and protein crosslinking and degradation. After fixation, PAXgene Tissue FIX is replaced with PAXgene Tissue STABILIZER Concentrate in the same container.

Note: Fixation rate depends on tissue type and size.

Preserved morphology
Stabilized samples can be embedded in paraffin for sectioning, and can be stained using standard protocols, such as hematoxylin/eosin (see H&E staining with the PAXgene Tissue System gives results comparable to formalin-fixed tissue) and immunohistochemistry (see Immunohistochemistry (IHC) staining with the PAXgene Tissue System gives comparable results to formalin-fixed tissue) for tissue sections with high staning intensity and clear morphological features.

Enhanced storage
When tissues are stored in PAXgene Tissue STABILIZER, nucleic acids, proteins and morphology of the tissue sample are stable for up to 7 days at room temperature (15–25°C) or 4 weeks at 2–8°C, depending on the type of tissue. Tissue samples can be stored in PAXgene Tissue STABILIZER for longer periods at –20°C (–15°C to –30°C) or –80°C (–65°C to –90°C) without negative effects on the morphology of the tissue or the integrity of the biomolecules.

Note: Specifications for fixation and storage conditions in PAXgene Tissue STABILIZER were determined using animal tissues. Storage at 2–8°C for more than 4 weeks must be validated for each tissue type.

Note: For storage at –20°C (–15°C to –30°C) or –80°C (–65°C to –90°C), use cryogenic vials with screw caps filled with PAXgene Tissue STABILIZER. For safety reasons, note that the PAXgene Tissue STABILIZER contains 70% ethanol by volume.

High-quality biomolecule stabilization
Nucleic acids and proteins of tissues stored in PAXgene Tissue STABILIZER or as PAXgene Tissue-fixed, paraffin-embedded (PFPE) samples are stabilized and can be purified using the corresponding PAXgene Tissue Kit for RNA and miRNA or for DNA. Use the Qproteome FFPE Tissue Kit for purification of proteins. Purified biomolecules are of high quality (see High-quality DNA from PFPE tissue with preserved morphology) and unmodified (see gDNA without chemical modifications can be used for demanding downstream applications). The purified biomolecules are highly suited for a range of demanding downstream applications (see High concordance of miRNA expression between total RNA isolated from PFPE and fresh-frozen tissueDetection of nondegraded, immunoreactive phosphoproteins from human PFPE tissue and RNA purified without chemical modification from PFPE tissue using the PAXgene Tissue RNA/miRNA Kit).
Principle
The methods for tissue fixation currently used in traditional histology are of limited use for molecular analysis. Fixatives that contain formaldehyde crosslink biomolecules and modify nucleic acids and proteins. Such crosslinks lead to nucleic acid degradation during tissue fixation, storage and processing. Since they cannot be removed completely, the resulting chemical modifications can lead to limitations in downstream applications, such as RT-PCR, qPCR and next-generation sequencing. The PAXgene Tissue FIX Container enables formalin-free fixation of tissue specimens. In combination with the PAXgene Tissue STABILIZER Concentrate, which enables formalin-free stabilization, tissue morphology and biomolecule integrity are preserved by avoiding destructive crosslinking and degradation found in formalin-fixed tissues (see The PAXgene Tissue FIX Container (50 ml) and PAXgene Tissue STABILIZER workflow).

Procedure
PAXgene Tissue FIX Containers are single-chamber containers prefilled with 50 ml of PAXgene Tissue FIX. PAXgene Tissue FIX Containers can accommodate 4 standard tissue cassettes (not provided), which can hold tissue samples with a maximum size of 4 x 15 x 15 mm. PAXgene Tissue FIX Containers also offer the possibility for direct fixation of larger tissue samples with a maximum size of 20 x 20 x 20 mm. PAXgene Tissue FIX rapidly penetrates and fixes the tissue. After fixation for 2 to 72 hours, depending on tissue size, PAXgene Tissue FIX is removed and replaced by PAXgene Tissue STABILIZER within the same container. Stabilized samples can be embedded in paraffin for histological studies, and nucleic acids and proteins can be isolated from the stabilized samples before or after embedding in paraffin.
Applications
PAXgene Tissue-fixed (PF) and PAXgene Tissue-fixed, paraffin-embedded (PFPE) tissue samples can be used for:
  • Pathological staining, including hematoxylin & eosin (H&E), periodic acid schiff (PAS), resorcin fuchsin, sirius red and Gomori
  • Immunohistochemical staining
  • in situ hybridization

 

Nucleic acids purified from PF and PFPE tissue samples can be used for demanding downstream applications, including:
  • RNA and miRNA profiling
  • Long-range and multiplex PCR
  • Next-generation sequencing

 

Proteins purified from PF and PFPE tissue samples can be used in a range of downstream applications, including:
  • Western blotting
  • Reverse-phase protein microarrays
  • MALDI imaging mass spectrometry
  • 2-D gel electrophoresis

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Brochures & Guides (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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FAQs (34)
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Scientific Posters (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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Kit Handbooks (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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Supplementary Protocols (16)

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