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PAXgene Tissue Container

For collection, fixation and stabilization of tissues

  • Preservation of both morphology and biomolecules
  • Fixation and paraffin embedding without formalin
  • Improved molecular results from fixed tissues
  • Tissues can be stored and archived for later processing
  • RNA, miRNA, DNA and/or proteins from one sample

 

Feature Specification
Container Dual-chamber container
Additives PAXgene Tissue FIX and PAXgene Tissue STABILIZER
Quantity 10 containers
Specimen size Up to 4 x 15 x 15 mm
Fixation time 2–24 h
Length of stabilization up to 7 days at room temperature (15–25°C)
up to 4 weeks at 2–8°C
long-term at –20°C or –80°C**
Archiving options for PFPE* short-term at room temperature or 2–8°C
long-term at –20°C or –80°C**

* PAXgene Tissue fixed, paraffin-embedded
** Long-term storage studies are on-going.

 

Intended Use

The PAXgene Tissue Container is intended for the collection, fixation, and stabilization of human tissue specimens. The container is intended to be used as part of the PAXgene Tissue Systems, which additionally include the PAXgene Tissue RNA Kit, PAXgene Tissue miRNA Kit, and PAXgene Tissue DNA Kit for the purification of RNA, total RNA (including miRNA), and DNA, respectively. Supplementary protocols are available for protein purification and other applications.

For research use only. Not for use in diagnostic procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease. The performance characteristics of this product have not been fully established.

The PAXgene Tissue Container Workflow
Dual-chamber container for fixation and stabilization of human tissue or cancer
H&E staining with the PAXgene Tissue System gives comparable results to formalin-fixed tissue.
Human tissue samples were divided into two sub-samples. One sub-sample was fixed with a PAXgene Tissue Container, and the other half was fixed in neutral-buffered formalin. The fixed tissues were [...]
Immunohistochemistry (IHC) staining with the PAXgene Tissue System gives comparable results to formalin-fixed tissue.
Human palatine tonsil tissue was fixed in PAXgene Tissue Containers or with neutral-buffered formalin and embedded in paraffin. Primary antibodies to the indicated antigens were linked to a [...]
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 [...]
DNA without chemical modifications 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., J Mol Diagn. 2012). (A) Multiplex PCR of [...]
High-quality RNA from PFPE tissue with preserved morphology.
(A) Hematoxylin and eosin (H&E) staining of PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue of different organs and (B) RNA from sections of rat PFPE. Electropherogram and RNA integrity [...]
RNA from PFPE tissue without chemical modifications
RNA from PFPE tissue without chemical modifications.
SYBR Green real-time RT-qPCR with 10 ng RNA from cryopreserved, formalin-fixed, paraffin-embedded (FFPE), or PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue (modified according to Groelz et [...]
Efficient purification of miRNA from PFPE tissue with the PAXgene Tissue miRNA Kit.
RNA purification from sections of six different PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue types using the PAXgene Tissue RNA Kit or PAXgene Tissue miRNA Kit. Amplification in a [...]
High concordance of miRNA expression between total RNA isolated from PFPE and flash-frozen tissue.
miRNA 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 [...]
Detection of nondegraded, immunoreactive phosphoproteins from human clinical PFPE tissue.
Non-malignant human uterus, breast, prostate and bladder specimens were divided into three samples and either flash-frozen in liquid nitrogen (cryo), PAXgene Tissue fixed, paraffin-embedded (PFPE), [...]
Protein extracted from PFPE tissue is suitable for two-dimensional gel electrophoresis.
Non-malignant human duodenum tissue was divided into three samples and either flash-frozen in liquid nitrogen (cryo), PFPE, or FFPE. Proteins from cryo and PFPE tissue were extracted in 2D buffer (30 [...]
Automation on the QIAcube: Significant hands-on time savings compared to the manual purification.
Comparison of sample preparation times between manual and QIAcube procedures for preparation of 12 PAXgene Tissue fixed, paraffin-embedded (PFPE) samples. Hands-on time without incubation times.
DNA 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 tissue, processed with the PAXgene Tissue DNA kit on the QIAcube in one single run. DNA [...]
Absorbance ratio 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. [...]
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. 300 [...]
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 eight different PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicates with the PAXgene Tissue DNA kit on the [...]
Comparison of manual and automated procedure: Absorbance ratio of sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue.
Ratio of absorbance at 260 and 280 nm of 3 sections (thickness 10 µm) each of eight different PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicates with the [...]
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 eight different PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicates with the PAXgene Tissue [...]
RNA yield of high-quality, high molecular weight RNA 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, RNA rich and RNA poor tissue, processed with the PAXgene Tissue RNA kit on the QIAcube. RNA yield per 10 mg [...]
Absorbance ratio of high-quality, high molecular weight RNA 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, RNA rich and RNA poor tissue, processed with the PAXgene Tissue RNA kit on the QIAcube in one single run. [...]
RIN value of high-quality, high molecular weight RNA 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, RNA rich and RNA poor tissue, processed with the PAXgene Tissue RNA kit on the QIAcube in one single run. RNA [...]
Comparison of manual and automated procedure: RNA yield from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue.
RNA yield from 5 sections (thickness 10 µm) each of eight different PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicates with the PAXgene Tissue RNA kit on the [...]
Comparison of manual and automated procedure: Absorbance ratio from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue.
Absorbance ratio at 260 and 280 nm of high-quality RNA from 5 sections (thickness 10 µm) each of eight different PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue types processed in [...]
Comparison of manual and automated procedure: RIN value from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue.
RNA from 5 sections (thickness 10 µ) each of eight different PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicates with the PAXgene Tissue RNA kit on the QIAcube [...]
Comparison of manual and automated procedure: Cycle treshold values from a reverse transcription and amplification from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue.
RNA from 5 sections (thickness 10 µ) each of eight different PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue types processed in triplicates with the PAXgene Tissue RNA kit on the QIAcube [...]
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Performance

PAXgene Tissue Containers are dual-cavity containers prefilled with 2 reagents. PAXgene Tissue FIX rapidly penetrates and fixes tissues, preserving tissue morphology and biomolecules. Fixation is comparable to formalin fixation, but avoids destructive nucleic acid and protein crosslinking and degradation. After fixation, tissue is transferred to PAXgene Tissue STABILIZER in the same container.

 

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 comparable results 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 staining 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 even be stored in the 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.

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 PAXgene Tissue STABILIZER contains 70% ethanol by volume.

Note: Specifications for fixation and storage conditions in PAXgene Tissue FIX and PAXgene Tissue STABILIZER were determined using animal tissues.

 

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, miRNA, or DNA, or the Qproteome FFPE Tissue Kit. Purified biomolecules are of high quality (see High-quality DNA from PFPE tissue with preserved morphology and High-quality RNA from PFPE tissue with preserved morphology) and unmodified (see DNA without chemical modifications can used for demanding downstream applications and RNA from PFPE without chemical modifications). 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 flash-frozen tissue, Detection of nondegraded, immunoreactive phosphoproteins from human clinical PFPE tissue and Protein extracted from PFPE tissue is suitable for two-dimensional gel electrophoresis).

 

Principle

Tissue fixation methods used in traditional histology are of limited use for molecular analysis. Fixatives containing formaldehyde cross-link biomolecules and modify nucleic acids and proteins, leading to degradation of analytes. Additionally, since cross-links cannot be removed completely, the resulting chemical modifications can cause inhibition in sensitive downstream applications such as quantitative PCR or RT-PCR. The PAXgene Tissue Container enables formalin-free fixation and stabilization of tissue specimens, which preserves tissue morphology and biomolecule integrity by avoiding destructive cross-linking and degradation found in formalin-fixed tissues.

 

Procedure

PAXgene Tissue Containers are dual-chamber containers prefilled with 2 reagents. Chamber 1 contains PAXgene Tissue FIX and chamber 2 contains PAXgene Tissue STABILIZER. Tissue samples with a maximum size of 4 x 15 x 15 mm are placed into a standard tissue cassette and fixed in chamber 1 for 2–24 hours. PAXgene Tissue FIX rapidly penetrates and fixes the tissue. After fixation, the tissue cassette is transferred to PAXgene Tissue STABILIZER in the chamber 2. Stabilized samples can be embedded in paraffin for histological studies. Nucleic acids and proteins can be isolated from the stabilized samples before or after embedding in paraffin.

Note: Fixation rates depend on tissue type and size.

 

Applications

PAXgene Tissue fixed (PF) and PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue samples can be used for (see References, Tissue Atlas):

 

Nucleic acids purified from PF and PFPE tissue samples can be used for demanding downstream applications (see References, Technical Notes), 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 (see References, Technical Notes), including:

  • Western blotting
  • Reverse-phase protein microarrays
  • MALDI imaging mass spectrometry
  • 2-D gel electrophoresis

 

Handbooks (1)

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PAXgene Tissue Container Product Circular - For fixation and stabilization of tissue specimens 333 KB pdf

Supplementary Protocols (18)

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Purification of full-length proteins from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue 652 KB pdf
Purification of full-length proteins from sections of PAXgene Tissue fixed, cryo-embedded (PFCE) tissue 651 KB pdf
Purification of full-length proteins from PAXgene Tissue fixed and stabilized (PF) tissue samples 629 KB pdf
Purification of total RNA, including miRNA, from sections of PAXgene Tissue fixed, cryo-embedded (PFCE) tissue placed directly into a microcentrifuge tube 621 KB pdf
Preparation of PFPE tissue sections for use with in situ hybridization (ISH) staining assays 606 KB pdf
Simultaneous purification of genomic DNA and total RNA, including miRNA, from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue 85 KB pdf
Simultaneous purification of genomic DNA and total RNA from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue 84 KB pdf
Purification of full-length proteins from sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue cut directly from a block of PFPE tissue 93 KB pdf
Purification of full-length proteins from slide-mounted sections of PAXgene® Tissue fixed, paraffin-embedded (PFPE) tissue 93 KB pdf
Deparaffinization of PAXgene Tissue fixed, paraffin-embedded tissue sections with Deparaffinization Solution 86 KB pdf
Manual processing of tissue specimens treated with the PAXgene Tissue System 342 KB pdf
Cryo-embedding tissue specimens fixed and stabilized with the PAXgene Tissue System 1109 KB pdf
Preparation of sections from PAXgene® Tissue fixed, paraffin-embedded (PFPE) and PAXgene Tissue fixed, cryo-embedded (PFCE) tissues for manual or laser microdissection (LMD) 97 KB pdf
Purification of total RNA, including miRNA, from microdissected PAXgene® Tissue fixed, paraffin embedded (PFPE) and PAXgene Tissue fixed, cryoembedded (PFCE) tissues 152 KB pdf
Purification of total RNA from sections of PAXgene® Tissue fixed, cryo-embedded (PFCE) tissue placed directly into a microcentrifuge tube 81 KB pdf
Purification of total RNA from microdissected PAXgene® Tissue fixed, paraffin-embedded (PFPE) and PAXgene Tissue fixed, cryo-embedded (PFCE) tissues 161 KB pdf
Purification of genomic DNA from sections of PAXgene® Tissue fixed, cryo-embedded (PFCE) tissue placed directly into a microcentrifuge tube 81 KB pdf
Purification of genomic DNA from microdissected PAXgene® Tissue-fixed, paraffin-embedded (PFPE) and PAXgene Tissue-fixed, cryo-embedded (PFCE) tissues 132 KB pdf

Brochures (6)

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Histological Staining with the PAXgene Tissue System 6800 KB pdf
Protein Extraction from PFPE 587 KB pdf
miRNA Isolation with the PAXgene Tissue miRNA Kit 957 KB pdf
RNA Isolation with the PAXgene Tissue RNA Kit 2417 KB pdf
DNA Isolation with the PAXgene Tissue DNA Kit 2018 KB pdf
PAXgene Tissue System Brochure 4792 KB pdf

Technical information (10)

File sizeFormatDownload
Quantitative analysis of KRAS and BRAF mutational status in DNA from PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue using Pyrosequencing technology 1404 KB pdf
Yield, purity, and integrity of RNA purified from PAXgene Tissue fixed, paraffin embedded (PFPE) rat tissue 618 KB pdf
Vaccum sealing of fixed and stabilized tissue with a FoodSaver Vacuum Sealer for dry and safe transport 886 KB pdf
Simultaneous preservation of RNA and morphology in tissue samples fixed with PAXgene Tissue Fix and stored for up to 2 years in PAXgene Tissue Stabilizer at –20 or –80°C 1313 KB pdf
RNA stability in tissue samples, fixed and stabilized with the PAXgene Tissue System, for up to 7 days at 22°C or 2 months at 4°C 538 KB pdf
Morphology and RNA preservation in PAXgene Tissue fixed, paraffin-embedded tissue (PFPE) stored for 18 months at different temperatures 495 KB pdf
Influence on RNA yield and integrity of modifications to the processing protocol 648 KB pdf
Influence of formalin contamination during processing of PAXgene Tissue fixed, paraffin-embedded tissue (PFPE) on RNA yield, integrity, and performance in quantitative RT-PCR 677 KB pdf
Effect of epitope retrieval conditions on immunohistochemical staining of PFPE tonsil tissue with anti-human Ki-67 antigen (clone MIB-1) 742 KB pdf
Detection of PI3K mutational status in DNA from human breast cancer PFPE tissue using the PI3K Mutation Test Kit (QIAGEN) 1028 KB pdf

Safety Data Sheets (1)

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MSDS PAXgene Tissue Container Link

Posters (16)

File sizeFormatDownload
New Fixation Technology For Simultaneous Preservation of Morphology and Nucleic Acids in Tissue; Groelz et al., AMP 2008 529 KB pdf
Long-Term Storage of Tissue Specimen at –20°C to –80°C with Preservation of Morphology and Nucleic Acids within Frozen Tissue; Groelz et al., AMP 2009/2 693 KB pdf
Preservation of Histomorphology and Nucleic Acids in Human Breast Tumor Tissue with the New PAXgene Tissue System — A Study with Comparison to Formalin Fixation; Groelz et al., AMP 2009/1 957 KB pdf
Preservation of Gene Expression Profile and Histomorphology in Human Breast Tumor Tissue with the New PAXgene Tissue System; Groelz et al., AACR 2010 769 KB pdf
Morphological, Epigenomic and Mutational Analyses of PAXgene Tissue Fixed, Paraffin-Embedded (PFPE) Colorectal Cancer (CRC) Specimens - Comparison to Formalin Fixed, Paraffin-Embedded (FFPE) and Snap-Frozen Samples; Groelz et al., AMP 2010 1189 KB pdf
A New Tissue Fixative for Biomarker Discovery: Gene expression and miRNA in PAXgene® Tissue Fixed Paraffin-Embedded (PFPE) Colorectal Cancer (CRC) and Breast Cancer Tissue vs. Formalin Fixed Paraffin-Embedded (FFPE) Tissue; Groelz et al., 3rd Annual Oncology Biomarkers 2011 423 KB pdf
PAXgene Tissue: A New Fixation Technology for Simultaneous Preservation of Morphology and Nucleic Acids; Groelz et al., BRN Symposium 2011 555 KB pdf
Evaluation of the PAXgene® Tissue System: Preservation of morphology and gene expression in human melanoma; Hesse et al., AACR-NCI-EORTC 2011 997 KB pdf
PAXgene vs. Formalin Fixed Tissue: A Comparison of Tissue Morphology and RNA Quality; Groelz el al., BRN Symposium 2012 5200 KB pdf
Preservation of Morphology and Biomolecules Within Tissue Stored for Three Years at -80°C in PAXgene Tissue Stabilizer Reagent; Groelz et al., ISBER 2012 912 KB pdf
PAXgene Tissue Fixation Technology for Simultaneous Preservation of Morphology and Biomolecules; Groelz et al., ECP 2012 1231 KB pdf
DNA quality measurement and somatic mutation profiling in PAXgene tissue samples with qBiomarker Somatic Mutation PCR Arrays; Long et al., ADAPT 2012 1223 KB pdf
RNA and Morphology Preservation after 5 Years storage at -20 and 3 years at -80°C; Groelz et al., GTEx Meeting 2014 909 KB pdf
Stability of Nucleic Acids in Archived Formalin and PAXgene Tissue Fixed, Paraffin-embedded Blocks of Tissue; Groelz et al., ECP 2014 2617 KB pdf
RT-PCR Performance of RNA Obtained From Archived FFPE and PFPE Blocks of Tissue; Groelz et al., AMP 2014 7568 KB pdf
RT-PCR Performance of RNA Obtained From Archived FFPE and PFPE Blocks of Tissue; Groelz et al., GTEx Meeting 2015 7568 KB pdf

References (24)

File sizeFormatDownload
Feature-based analysis of mouse prostatic intraepithelial neoplasia in histological tissue sections. Ruusuvuori P. et al., J Pathol Inform. 2016 Jan 29;7:5 Link
Pathogen Inactivating Properties and Increased Sensitivity in Molecular Diagnostics by PAXgene, a Novel Non-Crosslinking Tissue Fixative. Loibner M. et al., PLoS One. 2016 Mar 14;11(3):e0151383. Link
Protocol for HER2 FISH determination on PAXgene-fixed and paraffin-embedded tissue in breast cancer. Oberauner-Wappis L et al., Int J Exp Pathol. 2016 Apr;97(2):202-6 Link
The focus on sample quality: Influence of colon tissue collection on reliability of qPCR data. Korenkova V et al., Sci Rep. 2016 Jul 7;6:29023. doi: 10.1038/srep29023 Link
A Critical Evaluation of the PAXgene Tissue Fixation System:  Morphology, Immunohistochemistry, Molecular Biology, and Proteomics. Mathieson W. et al., Am J Clin Pathol. 2016 Jul;146(1):25-40. Link
Assessment of PAXgene Fixation on Preservation of Morphology and Nucleic Acids in Microdissected Retina Tissue. Liu Y and Edward DP, Curr Eye Res. 2016 Jul 13:1-7. Link
Comprehensive DNA Methylation and Extensive Mutation Analyses of HER2-Positive Breast Cancer; Yamaguchi et al. Oncology. 2015;88(6):377-84 Link
Surgical Specimens of Colorectal Cancer Fixed with PAXgene Tissue System Preserve High-Quality RNA; Hara K. et al., Biopreserv Biobank. 2015 Oct;13(5):325-34. Link
A Novel Approach to High-Quality Postmortem Tissue Procurement: The GTEx Project; Carithers et al., Biopreservation and Biobanking. October 2015, 13(5): 311-319. Link
Proteomic analysis of PAXgene-fixed tissues; Ergin B. et al., J Proteome Res. 2010 Oct 1;9(10):5188-96 Link
7568 KB pdf
Can we preserve morphology and nucleic acid integrity for molecular pathology? Mathieson et al., AMP 2010 746 KB pdf
Genotype-Tissue Expression (GTEx) Project Update, Jeff Struewing Sep 12, 2011 Link
Histological Assessment of PAXgene Tissue Fixation and Stabilization Reagents; Kap et al., PLoS ONE. 2011;6(11):e27704. Link
Biorepositories: Building better biobanks; Baker M., Nature 2012 Jun 6;486(7401):141-6. Link
A New Technology for Stabilization of Biomolecules in Tissues for Combined Histological and Molecular Analyses; Viertler C. et al., J Mol Diagn. 2012 Sep;14(5):458-66 Link
Non- Formalin Fixative versus Formalin-fixed Tissue: A Comparison of Histology and RNA Quality; Groelz et al., Exp Mol Pathol. 2013 Feb;94(1):188-94 Link
The PAXgene Tissue System Preserves Phosphoproteins in Human Tissue Specimens and Enables Comprehensive Protein Biomarker Research; Gündisch et al., PLoS One. 2013;8(3):e60638 Link
The Genotype-Tissue Expression (GTEx) project; Lonsdale et al., Nature Genetics 2013 May; 45, 580-585 Link
Preservation of nucleic acids and tissue morphology in paraffin-embedded clinical samples: comparison of five molecular fixatives; Staff et al., J Clin Pathol. 2013 Jun 8. [Epub ahead of print]. Link
Inactivation of Influenza A virus, Adenovirus, and Cytomegalovirus with PAXgene Tissue Fixative and Formalin; Kap et al., Biopreservation and Biobanking. August 2013, 11(4): 229-234. Link
Evaluation of colon cancer histomorphology: a comparison between formalin and PAXgene tissue fixation by an international ring trial; Gündisch et al., Virchows Arch 2014; DOI 10.1007/s00428-014-1624-4 Link
The Genotype-Tissue Expression (GTEx) Project: Linking Clinical Data with Molecular Analysis to Advance Personalized Medicine; Keen et al., J. Pers. Med. 2015, 5, 22-29 Link
Next-gen tissue: preservation of molecular and morphological fidelity in prostate tissue; Gillard M. et al., Am J Transl Res 2015;7(7):1227-1235 Link

FAQ (34)

    Tissue fixation and stabilization chemistry

    1. Which fixation method is used in the PAXgene Tissue System?

    The PAXgene Tissue System uses an acidic alcoholic fixation without formalin that does not result in the cross-linking of biomolecules.

    2. What is the fixation reagent of PAXgene Tissue FIX?

    The fixation reagent of PAXgene Tissue FIX contains alcohols and an acid, in addition to other stabilization agents.

    3. What is the stabilization reagent of PAXgene Tissue STABILIZER?

    The stabilization reagent of PAXgene Tissue STABILIZER contains alcohol and other stabilization agents.

    4. Why are two reagents used in the PAXgene Tissue System?

    The PAXgene Tissue System involves two processes: fixation and stabilization. PAXgene Tissue FIX provides rapid penetration and fixation that effectively stops all enzymatic activity throughout the tissue. PAXgene Tissue STABILIZER stops fixation and stabilizes the specimen for transportation and storage until processing.

    Tissue fixation and stabilization

    1. What is the maximum tissue size that can be fixed in a PAXgene Tissue Container?

    Fixation is performed within a standard tissue cassette. The maximum sample size is 4 x 15 x 15 mm, so that the tissue sample fits in the cassette without requiring physical pressure to close the lid. The cassette should leave no marks or grid impressions on the tissue.

    2. What is the maximum tissue size that can be fixed in a PAXgene Tissue FIX Container (50 ml)?

    Up to 4 standard tissue cassettes, each containing tissue samples with a maximum size of 4 x 15 x 15 mm, can be placed into a PAXgene Tissue FIX Container (50 ml). Alternatively, a single tissue sample with a maximum size of 20 x 20 x 20 mm can be fixed directly in a PAXgene Tissue FIX Container (50 ml). If using a larger tissue sample surrounded by fat (e.g., from a lymph node) or capsule (e.g., from kidney, liver or spleen tissue), partially cut into the tissue every 5 mm (lamination) to enhance permeation of the fixation reagent. If a sample is larger than recommended, the fast and even penetration of fixation reagent is compromised. In such cases, a reduction in the quality of tissue morphology and the integrity of nucleic acids may result.

    3. How long is the fixation time?

    For samples up to 4 x 15 x 15 mm fixed in a standard tissue cassette, incubate tissue specimen(s) at room temperature (15–25°C) for a minimum of 2 hours, but preferably 3–24 hours. For samples up to 20 x 20 x 20 mm fixed in the PAXgene Tissue FIX Container (50 ml), incubate for 6–48 hours, but preferably 8–24 hours. For biopsies with a thickness of 1 mm or less, fixation time can be reduced to 1 hour. Prolonged fixation times of up to 120 hours (e.g. over the weekend) is possible. Please note that fixation longer than the indicated times may lead to degradation of biomolecules.

    4. Which conditions are recommended for the storage of tissues in PAXgene Tissue STABILIZER?

    Depending on tissue type, standard storage conditions in PAXgene Tissue STABILIZER are a minimum of 2 hours and up to 7 days at room temperature (15–25°C) or up to 4 weeks at 2–8°C. For longer storage, samples can be kept in PAXgene Tissue STABILIZER at–15°C to –30°C or –65°C to –90°C. Long-term storage studies are ongoing. For the latest results, see the poster “RNA and Morphology Preservation after 5 years at –20°C and 3 years at –80°C” under the Resources tab.

    5. Are PAXgene Tissue Containers suitable for long-term storage at freezing temperatures?

    No. For storage at –15°C to –30°C or –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.

    Processing

    1. Is it possible to use a standard processor — the kind used routinely for formalin-fixed samples — for dehydration and paraffin infiltration of PAXgene Tissue treated samples?

    Yes. All processors commonly used for formalin-fixed samples can be used to produce PAXgene Tissue fixed, paraffin-embedded (PFPE) blocks of tissue. However, when processing PAXgene Tissue-treated specimens, do not use reagents contaminated with formalin. Residual formalin can lead to significant reduction of nucleic acid yield and integrity from PFPE tissue samples (see Technical Note “Influence of formalin contamination during processing of PAXgene Tissue fixed, paraffin-embedded tissue (PFPE) on RNA yield, integrity, and performance in quantitative RT-PCR”). Therefore, always use separate batches of reagents (alcohol, xylene, or xylene substitutes) for processing PAXgene Tissue fixed and formalin-fixed samples.

    2. Is it possible to process formalin-fixed and PAXgene Tissue fixed samples together in one run?

    Parallel processing of formalin-fixed and PAXgene Tissue fixed samples can lead to reduction of nucleic acid yield and integrity from PAXgene Tissue fixed samples through formalin contamination of reagents.

    3. Is it necessary to clean a processor normally used for formalin-fixed tissue before using it with PAXgene Tissue fixed tissue?

    No. Special cleaning is not required. However, be sure to change the alcohol and xylene (or xylene substitute), and keep separate batches of reagents for processing PAXgene Tissue fixed samples. Reagents contaminated with formalin can lead to reduced nucleic acid yields and integrity.

    4. Is a special processing protocol needed when using the PAXgene Tissue System?

    To prevent biomolecule degradation during processing, dehydration must begin with at least 70–100% ethanol. We recommend using low-melting paraffin (melting point ≤56°C) and do not incubate samples in liquid paraffin for more than 3 hours.

    Processing protocols for the PAXgene Tissue System are listed in the appendices of the “PAXgene Tissue Container Product Circular” and “PAXgene Tissue FIX Container (50 ml) Product Circular”.

    5. Is it possible to integrate the PAXgene Tissue STABILIZER into tissue processing?

    Yes. PAXgene Tissue STABILIZER can be used to fill the first position of a tissue processor. See the appendices of the “PAXgene Tissue Container Product Circular” and “PAXgene Tissue FIX Container (50 ml) Product Circular” for processing protocols with integrated PAXgene Tissue STABILIZER. When the STABILIZER is included as the first step of a protocol, tissues can be transferred from PAXgene Tissue FIX directly into the first processing position.

    6. Is it possible to archive PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue blocks?

    PFPE tissue blocks can be stored at 2–25°C for short-term storage or transport. However, biomolecules in paraffin blocks will undergo slow chemical degradation. To better preserve morphology and biomolecule integrity in the paraffin-embedded tissue, store PFPE blocks at–15°C to –30°C. For the latest results on long-term storage of PFPE and formalin-fixed, paraffin-embedded (FFPE) tissue, see the poster “RT-PCR performance of RNA obtained from archived FFPE and PFPE blocks of tissue” under the Resources tab.

    7. Is it possible to embed PAXgene Tissue fixed and stabilized samples in Optimal Cutting Temperture (OCT) medium for freezing?

    Yes. PreAnalytiX has developed a workflow and protocols for cryo-embedding tissue specimens fixed and stabilized in either the PAXgene Tissue Container or the PAXgene Tissue FIX Container (50 ml). Supplementary protocols for generating PAXgene Tissue fixed, cryo-embedded (PFCE) tissues are available under the Resources tab.

    Compatibility with conventional pathology techniques

    1. Is the morphology after H&E staining comparable to formalin-fixed samples?

    Yes. Comparable morphology was observed in adjacent pieces from a tissue sample fixed either with neutral-buffered formalin or with the PAXgene Tissue System for a variety of human and animal tissue (Gündisch et al., Virchows Arch. 2014; Kap et al., PLoS ONE 6(11): e27704). Examples are provided in the Tissue Atlas. PAXgene Tissue treated specimens have a tendency to be more eosinophilic. If an identical staining pattern to formalin-fixed samples is required, the incubation time in eosin should be reduced.

    2. Are immunohistochemistry (IHC) assays developed for formalin-fixed, paraffin-embedded (FFPE) tissues compatible with PAXgene Tissue fixed, paraffin-embedded (PFPE) tissues?

    Most antibodies used in IHC assays were developed for use with formalin-fixed tissue and include steps for unmasking epitopes. When working with PAXgene Tissue treated specimens, test each antibody to determine if it is necessary to perform antigen-retrieval steps. In addition, it may be necessary to optimize antigen-retrieval steps or adjust antibody concentrations in PFPE tissue to achieve optimal staining intensities (see Technical Note “Effect of epitope retrieval conditions on immunohistochemical staining of PFPE tonsil tissue with anti-human Ki-67 antigen (clone MIB-1)” under the Resources tab). Examples for IHC staining of adjacent human tissue sections fixed with neutral-buffered formalin or with PAXgene Tissue reagents are provided in the Tissue Atlas.

    3. Can sections of PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue be used for other histochemical staining techniques, such as PAS?

    Human tissue samples treated with the PAXgene Tissue System were successfully used for periodic acid schiff (PAS), resorcin fuchsin, sirius red, and Gomori staining (Kap et al., PLoS ONE 6(11): e27704). However, to achieve the same staining intensities with both PFPE and formalin-fixed, paraffin-embedded (FFPE) samples, it may be necessary to adjust incubation times.

    4. Can PAXgene Tissue fixed, paraffin-embedded tissue be used for in situ hybridization?

    Yes, human tissue samples treated with the PAXgene Tissue System have been used successfully for fluorescence in situ hybridization (FISH) and chromogenic in situ hybridization (CISH). However, specimens treated with the PAXgene Tissue System are more sensitive to enzyme digestion compared to formalin-fixed samples. Enzyme digestion and other pretreatment steps may need to be optimized.

    Purification and quality of biomolecules from PAXgene Tissue treated samples

    1. Is it possible to use formalin-fixed, paraffin-embedded (FFPE) kits and protocols to isolate biomolecules from PAXgene Tissue fixed, paraffin-embedded (PFPE) tissues?

    No. Procedures developed for the extraction of biomolecules from FFPE tissues include prolonged proteinase K digestion and heating steps to remove chemical modifications introduced by formalin. Since the PAXgene Tissue System does not chemically modify biomolecules, these steps are not necessary and, in fact, lead to degradation of biomolecules. Instead, use dedicated PAXgene Tissue kits and supplementary protocols for extraction of biomolecules from PAXgene Tissue treated samples.

    2. What is the purity of nucleic acids extracted with the PAXgene Tissue kits?

    The PAXgene Tissue DNA, RNA, and miRNA Kits are based on proven QIAGEN technologies. Nucleic acids isolated with these kits are generally of high purity.

    On average, measurements of the A260/A280 ratio for DNA purified with the PAXgene Tissue DNA Kit are >1.7, and ratios for RNA and miRNA purified with the PAXgene Tissue RNA or miRNA Kits are both >1.8.

    For examples of RNA purity see Technical Note "Yield, purity, and integrity of RNA purified from PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue" under the Resources tab.

    3. What is the average RNA yield from PAXgene Tissue fixed, paraffin-embedded (PFPE) tissues?

    RNA yield depends on several parameters, such as tissue type, time from resection until fixation, fixation time (ideally 2 to 4 hours), processing protocol used and age and storage conditions of the PFPE block.

    In a study with PFPE tissue sections (area: 100 mm²;  thickness: 10 µm) median RNA yield from rat liver was 4.2 µg (n=58), from kidney 2.2 µg (n=58), from spleen 4.7 µg (n=58), from intestine 4.7 µg (n=58) and from lung 0.9 µg (n=58) (see Technical Note "Yield, purity, and integrity of RNA purified from PAXgene Tissue fixed, paraffin-embedded (PFPE) rat tissue" under the Resources tab).

    4. How well is RNA integrity preserved in PAXgene Tissue fixed, paraffin-embedded (PFPE) tissues?

    Similar to yield, RNA integrity depends on several parameters, such as tissue type, time from resection until fixation, fixation time, processing protocol and age and storage conditions of the PFPE block. For examples of RNA integrity values from rat tissues under ideal workflow conditions, see Groelz et al., Exp Mol Pathol. 2013 Feb; 94(1). For examples of RNA integrity from clinical samples, see Viertler et al., J Mol Diagn. 2012 Sep; 14(5).

    5. How well is DNA integrity preserved in PAXgene Tissue fixed, paraffin-embedded (PFPE) tissues?

    In contrast to DNA isolated from formalin-fixed, paraffin-embedded (FFPE) tissue, DNA from PFPE tissue exhibits high molecular weight. In most cases, a distinct 10 kb band is observed in electrophoretically separated DNA eluates. For an example, see Figure 2 in the Technical Note “Quantitative analysis of KRAS and BRAF mutational status in DNA from PAXgene Tissue fixed, paraffin-embedded (PFPE) tissue using Pyrosequencing technology“ under the Resources tab.

    6. What are the yield and integrity of nucleic acids isolated from blocks of PAXgene Tissue fixed, cryo-embedded (PFCE) tissues?

    DNA and RNA isolated from PFCE tissue specimens are of high quantity and quality, comparable to PFPE tissue.

    7. Are special kits and protocols required for the isolation of biomolecules from PAXgene TIssue fixed, cryo-embedded (PFCE) tissues?

    Regular PAXgene Tissue kits can be used for the isolation of RNA, miRNA and DNA from PFCE tissue. Supplementary protocols developed specifically for the extraction of biomolecules from PFCE samples are available under the Resources tab.

    8. Does the PAXgene Tissue RNA Kit co-purify small RNAs?

    Yes, small RNAs are co-purified. However, due to the purification chemistry, RNA molecules smaller than approximately 200 nucleotides are recovered with less efficiency. For purification of total RNA, including miRNA and other small RNA molecules of at least 18 nucleotides, use the PAXgene Tissue miRNA Kit.

    9. Can proteins be extracted from PAXgene Tissue fixed specimens?

    Yes. Supplementary protocols are available for the purification of full-length proteins from PAXgene Tissue fixed (PF) tissue and paraffin blocks using the Qproteome FFPE Tissue Kit (QIAGEN, cat. no. 37623). For more information, see the corresponding supplementary protocols under the Resources tab.

    10. Is it possible to microdissect PAXgene Tissue fixed, paraffin-embedded (PFPE) and PAXgene Tissue fixed, cryo-embedded (PFCE) tissues?

    Yes, supplementary protocols for generating sections from PFPE and PFCE tissue blocks for manual and laser microdissection are available under the Resources tab.

    11. Which kits and protocols can be used for the isolation of nucleic acids from microdissected PAXgene Tissue fixed, paraffin-embedded (PFPE) and PAXgene Tissue fixed, cryo-embedded (PFCE) specimens?

    Regular PAXgene Tissue kits can be used for the isolation of RNA, miRNA and DNA from microdissected PFPE and PFCE tissue. Supplementary protocols developed specifically for the extraction of biomolecules from microdissected PFPE and PFCE samples are available under the Resources tab.

    Molecular analysis of biomolecules purified from PAXgene Tissue treated samples

    1. What is the RT-PCR performance of RNA purified from PAXgene Tissue fixed, paraffin-embedded (PFPE) and PAXgene Tissue fixed, cryo-embedded (PFCE) tissues compared to RNA from snap frozen or formalin-fixed, paraffin-embedded (FFPE) tissues?

    The PAXgene Tissue fixation and stabilization chemistry is free of cross-linking reagents. RNA purified from PFPE and PFCE is free of chemical modifications and performs similarly or identically to RNA isolated from frozen tissue. For examples of the correlation of gene expression levels in snap frozen tissue, FFPE, and PFPE, see Figure 4 in Groelz et al., Exp Mol Pathol. 2013 Feb; 94(1) and Figure 3 in Viertler et al., J Mol Diagn. 2012 Sep; 14(5).

    2. What is the PCR performance of DNA purified from PAXgene Tissue fixed, paraffin-embedded (PFPE) and PAXgene Tissue fixed, cryo-embedded (PFCE) tissues compared to DNA from snap frozen or formalin-fixed, paraffin-embedded (FFPE) tissues?

    In contrast to FFPE, DNA purified from PFPE and PFCE is of high molecular weight and free of chemical modifications. In demanding downstream applications, such as multiplex or long-range PCR, it performs similarly or identically to DNA isolated from frozen tissue. For examples, see, Figure 5 in Viertler et al., J Mol Diagn. 2012 Sep; 14(5).

    3. How is the quality of proteins purified from PAXgene Tissue fixed and stabilized tissues?

    Proteins from PAXgene Tissue fixed, paraffin-embedded (PFPE) and PAXgene Tissue fixed, cryo-embedded (PFCE) tissues are non-degraded, immunoreactive and have been successfully investigated by western blot analysis, reverse-phase protein arrays, two-dimensional gel electrophoresis (2D-PAGE), enzyme-linked immunosorbent assay (ELISA) and MALDI imaging mass spectrometry.

ProductCatalog Nr.Price (USD)

PAXgene Tissue Container (10)

For collection, fixation, and stabilization of 10 tissue samples: 10 Prefilled Reagent Containers, containing PAXgene Tissue Fix and PAXgene Tissue STABILIZER Reagent.
765112156.00