PAXgene® Tissue Container Product Circular
Contents
1. This is not a complete list of suppliers and does not include many important vendors of biological supplies 10 PAXgene Tissue Container Product Circular 04 2013 Important Notes Storage and archiving of PFPE PAXgene Tissue fixed paraffin embedded tissue samples For short term storage or transport blocks of PFPE tissue can be stored at room temperature 1 5 25 6 or refrigerated temperature 2 25 C However biomolecules within the paraffin blocks will undergo slow chemical degradation For best preservation of morphology and maintenance of biomolecule integrity within the parattin embedded tissue store PFPE blocks at 20 C 15 C to 30 C Labeling of PAXgene Tissue Container To allow calculation of the exact length of fixation time record the date and time that tissue is placed in the PAXgene Tissue Container in the Fixation Date Time field on the Container label Figure 4 After transferring the specimen from PAXgene Tissue FIX into PAXgene Tissue STABILIZER record the date and time of the transfer in the Transfer Date Time field on the cap label T Spe Bu PAXgene Tissue Containet Figure 4 Cap label of a PAXgene Tissue Container PAXgene Tissue Container Product Circular 04 2013 11 Protocol Sample Fixation and Stabilization Starting material Starting material for tissue fixation should be a sample cut to a maximum size of 4 15 15 mm Important points before starting Do not use tiss2. concentration in PFPE tissue to achieve optimal staining intensities For the latest information on IHC protocols and staining conditions see the tissue atlas at www preanalytix com product catalog tissue tissue atlas PAXgene Tissue Container Product Circular 04 2013 19 References QIAGEN maintains a large up to date online database of scientific publications utilizing QIAGEN and PreAnalytiX products Comprehensive search options allow you to find the articles you need either by a simple keyword search or by specifying the application research area title etc For a complete list of references visit the QIAGEN Reference Database online at www qiagen com RefDB search asp or at www preanalytix com or contact QIAGEN Technical Services or your local distributor 20 PAXgene Tissue Container Product Circular 04 2013 Cat no 765112 765312 765512 765134 766134 21 Ordering Information Product Contents PAXgene Tissue For fixation and stabilization of tissue Container 10 specimens 10 Prefilled Reagent Containers containing PAXgene Tissue FIX and PAXgene Tissue STABILIZER Related products PAXgene Tissue FIX Container for fixation and stabilization of tissue specimens PAXgene Tissue FIX 10 Pretilled Reagent Containers Container 50 ml containing 50 ml of PAXgene Tissue FIX PAXgene Tissue STABILIZER for stabilization of tissue samples treated with PAXgene Tissue PAXgene Tissue 9 bottles of PAXg
3. 2 other successfully tested protocols 3 Immediately after the final paraffin incubation step embed the tissue sample into a block of paraffin For embedding use the same low melting paraffin used for infiltration 4 After hardening store paraffin blocks in a dry dark place Note The ideal storage temperature to preserve nucleic acid integrity and tissue morphology for paraffin embedded tissue is 20 C 15 C to 30 C 14 PAXgene Tissue Container Product Circular 04 2013 5 When ready for further analysis cut paraffin blocks with a microtome and transfer the sections onto the surface of a water bath heated to 40 C for 1 min IMPORTANT Do not heat the water bath above 40 C Stretching the sections on water with a temperature above 40 C overstretches morphological structures Do not leave the sections in the water bath for longer than 1 min 6 Dry sections overnight at room temperature 15 25 C PAXgene Tissue Container Product Circular 04 2013 15 Appendix A Generic Processing Protocol for an Automated Tissue Processor That Supports Preservation of Biomolecules in Specimens Treated with the PAXgene Tissue System Table 1 Generic processing protocol Media Alternatives 80 99 ethanol PAXgene Tissue STABILIZER 901 99 ethanol None mandatory 951 99 ethanol None mandatory 9996 ethanol None mandatory 9996 ethanol None mandatory 99 ethanol Isopropanol 99 ethanol Isopropanol Xylene Xylene substitutes
4. 5 25 60 for a minimum of 2 h but preferably 3 24 h Note Longer fixation periods may lead to degradation of biomolecules Note For biopsies with a thickness of 1 mm or less fixation time can be reduced to 30 60 min PAXgene Tissue Container Product Circular 04 2013 l 12 After fixation unscrew the cap and lift up the screw cap rack assembly with tissue cassette from chamber 1 Position the rack tissue cassette directly above chamber 2 containing PAXgene Tissue STABILIZER and carefully submerge Tighten the screw cap Note Make sure that the cap is correctly seated on the rim of the PAXgene Tissue Container before tightening the cap Store or transport tissue specimen in PAXgene Tissue STABILIZER Note Standard storage conditions are up to 7 days at room temperature or for up to 4 weeks at 2 8 C depending on tissue type see footnote on page 7 Samples can be frozen at 20 C 15 C to 30 C or at 80 C 65 C to 90 C For freezing we recommend transferring the tissue sample from the tissue cassette and placing it into a screw cap cryovial not supplied filled with PAXgene Tissue STABILIZER Note A minimum incubation time of 2 h in PAXgene Tissue Stabilizer is recommended before processing and embedding in paraffin PAXgene Tissue Container Product Circular 04 2013 13 8 9 10 Protocol Sample Processing Paraffin Embedding and Sectioning Important points before starting e Refer to Ap
5. Luxembourg Orders 8002 2076 Fax 8002 2073 Technical 8002 2067 Mexico Orders 01 800 7742 639 Fax 01 800 1122 330 Technical 01 800 7742 639 The Netherlands Orders 0800 0229592 Fax 0800 0229593 Technical 0800 0229602 Norway Orders 800 18859 Fax 800 18817 Technical 800 18712 Singapore Orders 65 67775366 Fax 65 67785177 Technical 65 67775366 Spain Orders 91 630 7050 Fax 91 630 5145 Technical 91 630 7050 Sweden Orders 020 790282 Fax 020 790582 Technical 020 798328 Switzerland Orders 055 254 22 11 Fax 055 254 22 13 Technical 055 254 22 12 UK Orders 01293 422 911 Fax 01293 422 922 Technical 01293 422 999 USA Orders 800 426 8157 Fax 800 718 2056 Technical 800 DNA PREP 800 362 7737 www qgiagen com www PreAnalytiX com Argentina Uruguay and Paraguay Orders 011 4551 7100 Australia Orders 1 800 656 100 Fax 1 800 656 110 Austria Orders 43 1 7063660 Fax 43 1 7063660 11 Belgium Orders 32 53720408 Fax 32 53720558 Brazil Orders 0800 055 5654 Canada Orders 800 268 5430 Fax 800 565 0897 Denmark Orders 45 43 43 45 66 Fax 45 43 43 41 66 East Europe Middle East amp Africa EMA Orders 971 4 3379525 Fax 971 4 03379551 Finland Orders 358 9 88 70 780 Fax 358 9 88 70 7817 France Orders 33 476 683636 Fax 33 476 683693 Germany Orders 49 6221305553 Fax 49 6221305377 Italy Orders 390248204775 Fax 390248204817 Technical 390248240264 The Netherlands O
6. since the performance characteristics of these Containers have not been validated for any specific application The performance characteristics of this product have not been fully established 4 PAXgene Tissue Container Product Circular 04 2013 Safety Information When working with chemicals always wear a suitable lab coat disposable gloves and protective goggles For more information please consult the appropriate satety data sheets SDSs These are available online in convenient and compact PDF format at www qiagen com satety where you can find view and print the SDS for each PreAnalytiX Kit and kit component The following risk and safety phrases apply to components of the PAXgene Tissue Container PAXgene Tissue FIX QA F T Contains methanol acetic acid toxic irritant highly flammable Risk and safety phrases R11 23 24 25 36 38 39 23 24 25 57 16 36 37 39 45 PAXgene Tissue STABILIZER Contains ethanol highly flammable Risk and safety phrases R11 6 24 hour emergency information Emergency medical information in English French and German can be obtained 24 hours a day from Poison Information Center Mainz Germany Tel 49 6131 19240 Quality Control In accordance with QIAGEN s ISO certified Total Quality Management System each lot of the PAXgene Tissue Container is tested against predetermined specifications to ensure consistent product quality R11 Highly flammable R23 24 25 Toxic by i
7. DNA RNA or miRNA and the PAXgene Tissue supplementary protocols www preanalytix com for protein purification and other applications Fixation rates and stabilization times depend on type and size of tissue Specifications for tissues size fixation and storage conditions in PAXgene Tissue FIX and PAXgene Tissue STABILIZER were determined using animal tissues samples t For the latest results on long term storage see technical notes at www preanalytix com PAXgene Tissue Container Product Circular 04 2013 7 Excise and cut tissue to size place into a standard tissue cassette 4x15x15mm Remove screw cap rack assembly Insert the lower edge of the tissue cassette into the bottom edge of the rack Attach tissue cassette to the rack Submerge the rack holding the tissue cassette into chamber 1 containing PAXgene Tissue FIX Screw the cap into place Allow fixation for 2 to 24 hours Figure 1 Fixation within Chamber 1 of the PAXgene Tissue Container 8 PAXgene Tissue Container Product Circular 04 2013 After fixation remove rack with tissue cassette trom chamber 1 Submerge rack holding tissue cassette into chamber 2 containing PAXgene Tissue STABILIZER Screw the cap into place the container is ready for transport After processing and paraffin embedding the block of PAXgene NAT Tissue fixed paraffin embedded m PFPE tissue is ready for sectioning Figure 2 Stabili
8. PAXgene Tissue Container Product Circular For fixation and stabilization of tissue specimens For research use only Not for use in diagnostic procedures April 2013 Trademarks PAXgene PreAnalytiX PreAnalytiX GmbH QIAGEN MinElute Qproteome QIAGEN Group Paraplast X tra Leica Biosystems Inc Thermo Fisher Scientific Thermo Fisher Scientific Thermo Fisher Scientific Inc Limited License Agreement Use of this product signifies the agreement of any purchaser or user of the PAXgene Tissue Container to the following terms The PAXgene Tissue Container may be used solely in accordance with the PAXgene Tissue Container Product Circular and for use with components contained in the Product only PreAnalytiX grants no license under any of its intellectual property to use or incorporate the enclosed components of this Product with any components not included within this Product except as described in the PAXgene Tissue Container Product Circular and additional protocols available at www preanalytix com Other than expressly stated licenses PreAnalytiX makes no warranty that this Product and or its use s do not infringe the rights of third parties This Product and its components are licensed for one time use and may not be reused refurbished or resold PreAnalytiX specifically disclaims any other licenses expressed or implied other than those expressly stated The purchaser and user of the Product agree not to take or per
9. book or user manual PreAnalytix and QIAGEN kit handbooks and user manuals are available at www giagen com or can be requested from QIAGEN Technical Services or your local distributor 22 PAXgene Tissue Container Product Circular 04 2013 PreAnalytiX Worldwide PreAnalytiX products are distributed by QIAGEN and BD companies Australia Orders 03 9840 9800 Fax 03 9840 9888 Technical 1 800 243 066 Austria Orders 0800 28 10 10 Fax 0800 28 10 19 Technical 0800 28 10 11 Belgium Orders 0800 79612 Fax 0800 79611 Technical 0800 79556 Brazil Orders 0800 557779 Fax 55 11 5079 4001 Technical 0800 557779 Canada Orders 800 572 9613 Fax 800 713 5951 Technical 800 DNA PREP 800 362 7737 China Orders 021 3865 3865 Fax 021 3865 3965 Technical 800 988 0325 Denmark Orders 80 885945 Fax 80 885944 Technical 80 885942 Finland Orders 0800 914416 Fax 0800 914415 Technical 0800 914413 France Orders 01 60 920 926 Fax 01 60 920 925 Technical 01 60 920 930 Offers 01 60 920 928 Germany Orders 02103 29 12000 Fax 02103 29 22000 Technical 02103 29 12400 Hong Kong Orders 800 933 965 Fax 800 930 439 Technical 800 930 425 Ireland Orders 1800 555 049 Fax 1800 555 048 Technical 1800 555 061 Italy Orders 02 33430411 Fax 02 33430426 Technical 800 787980 Japan Telephone 03 5547 0811 Fax 03 5547 0818 Technical 03 5547 0811 Korea South Orders 1544 7145 Fax 1544 7146 Technical 1544 7145
10. cation of RNA DNA or total RNA including miRNA In addition supplementary protocols tor protein purification and other applications are available at www preanalytix com Principle and procedure PAXgene Tissue FIX rapidly penetrates and fixes tissue with a fixation rate of approximately 1 mm in 30 minutes In research applications the reagent preserves morphology and biomolecules without the destructive cross linking and degradation tound with formalin tixed tissues The fixation process is stopped by transferring the tissue specimen into PAXgene Tissue STABILIZER After fixation tissues can be stored directly in PAXgene Tissue STABILIZER for the short or long term used for extraction of nucleic acids or proteins or processed and embedded in paraffin for further analysis Sections of PAXgene Tissue fixed paraffin embedded PFPE tissue can be used for histological studies or extraction of nucleic acids or proteins Purification of RNA total RNA including miRNA or DNA from PAXgene Tissue fixed and stabilized tissue samples requires the use of one of the PAXgene Tissue Kits for RNA miRNA or DNA Purification of protein requires the Qproteome FFPE kit QIAGEN PAXgene Tissue reagents in prefilled containers and PAXgene Tissue kits provide a complete preanalytical solution for collection fixation and stabilization of tissue and purification of high quality nucleic acids for molecular research analysis Tissue penetration and fi
11. ene Tissue STABILIZER STABILIZER Concentrate concentrate for 4 liters of PAXgene 150 ml Tissue STABILIZER PAXgene Tissue RNA Kit for purification of total RNA from tissues fixed and stabilized using the PAXgene Tissue System PAXgene Tissue RNA Kit For 50 RNA preps PAXgene RNA 50 MinElute Spin Columns PAXgene Shredder Spin Columns Processing Tubes Microcentrituge Tubes Carrier RNA RNase Free DNase and RNase Free Buffers to be used in conjunction with PAXgene Tissue Containers PAXgene Tissue miRNA Kit for purification of total RNA including miRNA from tissue samples fixed and stabilized using the PAXgene Tissue System PAXgene Tissue miRNA For 50 RNA preps PAXgene RNA Kit 50 MinElute Spin Columns PAXgene Shredder Spin Columns Processing Tubes Microcentrifuge Tubes Carrier RNA RNase Free DNase and RNase Free Buffers to be used in conjunction with PAXgene Tissue Containers PAXgene Tissue Container Product Circular 04 2013 Product Contents Cat no PAXgene Tissue DNA Kit for purification of DNA from tissues samples fixed and stabilized using the PAXgene Tissue System PAXgene Tissue DNA For 50 DNA preps PAXgene DNA Mini 767134 Kit 50 Spin Columns Processing Tubes Microcentrituge Tubes Carrier RNA and Butters to be used in conjunction with PAXgene Tissue Containers For up to date licensing information and product specitic disclaimers see the respective PreAnalytiX or QIAGEN kit hand
12. ess of the tissue specimen PAXgene Tissue Container Product Circular 04 2013 17 Vacuum 0 5 bar 0 5 bar 0 5 bar 0 5 bar 0 5 bar 0 5 bar 0 5 bar 0 5 bar 0 5 bar Temperature 15 25 C 15 25 C 15 25 C 15 25 C 15 25 C 15 25 C 15 25 C 15 25 C 15 25 C 50 C Table 3 Processing protocol B Time 15 min 12h 30 min 30 60 min 30 60 min 30 60 min 30 60 min 30 60 min 30 60 min 30 60 min 30 60 min 30 60 min 60 min Media 8096 ethanol 90 ethanol 9596 ethanol 9996 ethanol 9996 ethanol Isopropanol Isopropanol Xylene Xylene 1 1 mixture of Paraplast X tra and Xylene Paraplast X tra Paraplast X tra 2 3 4 6 8 9 Optimal incubation times depend on the thickness of the tissue specimen PAXgene Tissue Container Product Circular 04 2013 18 Appendix 6 Optimizing Immunohistochemistry IHC Assays with Sections of PFPE PAXgene Tissue Fixed Paraffin Embedded Tissue In contrast to formalin or other fixation reagents containing aldehydes the PAXgene Tissue System does not cause cross linking of biomolecules Therefore it may not be necessary to unmask epitopes for immunohistochemistry assays either by heating or proteolytic digestion It is important to note however to note that since many antibodies used in immunohistochemical assays were developed for use with formalin fixed tissue is often necessary to optimize antigen retrieval steps and or to adjust antibody
13. may be used but do not use clearing agents based on D limonene Xylene Xylene substitutes may be used but do not use clearing agents based on D limonene Parattin with melting 1 1 mixture of paraffin and xylene point 54 58 C Paraffin with melting None mandatory point 54 58 C Paraffin with melting If only 2 stations that can be heated are point 54 58 C available omit this step When processing specimens fixed in PAXgene Tissue Containers do not use reagents contaminated with formalin Formalin contamination in the alcohol used for sample processing can lead to significant reduction of nucleic acid and protein yield t Use filtered or deionized water for preparation of 80 90 or 95 ethanol 16 PAXgene Tissue Container Product Circular 04 2013 Appendix 5 Examples of Automated Processing Protocols Successfully Used for Specimens Treated with the PAXgene Tissue System Table 2 Processing protocol A Media Time Temperature Vacuum PAXgene Tissue Up to 7 days 18 22 C STABILIZER 99 ethanol 30 60 min 18 22 C 99 ethanol 30 60 min 18 22 C 99 ethanol 30 60 min 18 22 C 99 ethanol 30 60 min 18 22 C 99 ethanol 30 60 min 18 22 C lsopropanol 30 60 min 18 22 C lsopropanol 30 60 min 18 22 C Xylene 30 60 min 18 22 C Xylene 30 60 min 18 22 C Paraplast X tra 30 60 min 56 C Paraplast X tra 90 min 56 C SO CON WB Oo d W N BEA N Oo Optimal incubation times depend on the thickn
14. mit anyone else to take any steps that could lead to or facilitate any acts prohibited above PreAnalytiX may enforce the prohibitions of this Limited License Agreement in any Court and shall recover all its investigative and Court costs including attorney fees in any action to enforce this Limited License Agreement or any of its intellectual property rights relating to the Product and or its components 1 OO N For updated license terms see www preanalytix com 2013 PreAnalytiX GmbH all rights reserved PreAnalytiX Company PreAnalytiX GmbH Feldbachstrasse CH 8634 Hombrechtikon Switzerland www preanalytix com PreAnalytiX Distributors PreAnalytiX products are manufactured for PreAnalytiX by QIAGEN or BD and are distributed for PreAnalytiX by QIAGEN or BD Products cannot be ordered from PreAnalytiX GmbH Please see the last page for contact information for your local PreAnalytiX distributor Contents Kit Contents 4 Symbols 4 Shipping and Storage 4 Intended Use 4 Safety Information 5 Quality Control 5 Introduction 6 Principle and procedure 6 Description of protocols 7 Sample collection and stabilization with the PAXgene Tissue Container 7 Equipment and Reagents to Be Supplied by User 10 Important Notes 11 Storage and archiving of PFPE PAXgene Tissue fixed paraftin embedded tissue samples 11 Labeling of PAXgene Tissue Container 11 Protocol Sample Fixation and Stabilization 12 Protocol Sample P
15. nhalation in contact with skin and if swallowed R36 38 Irritating to eyes and skin R39 23 24 25 Toxic danger of very serious irreversible effects through inhalation in contact with skin and if swallowed S7 Keep container tightly closed 16 Keep away from sources of ignition No smoking S36 37 39 Wear suitable protective clothing gloves and eye face protection 45 In case of accident or If you feel unwell seek medical advice immediately show the label where possible PAXgene Tissue Container Product Circular 04 2013 5 Introduction The methods for tissue fixation currently used in traditional histology are of limited use for molecular analysis Fixatives that contain formaldehyde cross link biomolecules and modify nucleic acids and proteins Such cross links lead to nucleic acid degradation during tissue fixation storage and processing Since they cannot be removed completely the resulting chemical modifications can cause inhibition in downstream applications such as quantitative PCR or RT PCR To enable both molecular and traditional pathology testing from the same specimen a method is needed to stabilize molecular content and preserve morphology PreAnalytiX has developed the PAXgene Tissue System to meet this need The system consists of a fixation reagent PAXgene Tissue FIX a stabilization reagent PAXgene Tissue STABILIZER prefilled containers for tissue collection storage and transportation and kits for purifi
16. pendices A and B for detailed processing protocols pages 16 18 e Do not begin tissue processing with either water or with ethanol at dilutions less than 8096 e Denatured ethanol can be used for processing e Do not reuse processing reagents previously contaminated with formalin even in trace amounts as this can lead to significant reductions in DNA and RNA yield and quality e Tissue samples fixed and stabilized in PAXgene Tissue reagents can remain in the first station of the tissue processor in 80 9996 ethanol for up to 12 h e For paraffin infiltration the liquid paraffin should be held at 2 C above its melting point Do not let the incubation temperature of the paraffin exceed 60 C and do not incubate in liquid paraffin for longer than 3 h Extensive incubation times above 60 C lead to degradation of RNA Procedure 1 Transfer the tissue cassette into the first station of a tissue processor filled with 80 99 ethanol to start dehydration Note Reagents used for processing must be free of formalin contamination Note In case a tissue processor with formalin free reagents is not available specimens treated with the PAXgene Tissue System can be processed manually A supplementary protocol for manual processing is available at the PAXgene Tissue Resources section of www preanalytix com 2 Follow a protocol for paraffin embedding See Appendix A on page 16 for a generic protocol or Appendix B on page 17 for examples of
17. rders 31 20 6545 716 Fax 31 20 5829 421 New Zealand Orders 0800 572 468 Fax 0800 572 469 Spain Orders 34 91 848 8115 Fax 34 91 848 8104 Sweden Orders 46 8 775 51 00 Fax 46 8 775 51 95 Switzerland Orders 41 61 4852222 Fax 41 61 4852200 UK Orders 44 1 865 781 666 Fax 44 1 865 781 528 USA Orders 888 237 2762 Fax 800 847 2220 Technical 800 631 0174 www bd com www PreAnalytiX com PAXgene Tissue Container Product Circular 04 2013 23 69 PreAnalytiX 1075949 04 2013
18. rocessing Paraffin Embedding and Sectioning 14 Appendix A Generic Processing Protocol for an Automated Tissue Processor That Supports Preservation of Biomolecules in Specimens Treated with the PAXgene Tissue System 16 Appendix B Examples of Automated Processing Protocols Successfully Used for Specimens Treated with the PAXgene Tissue System 17 Appendix C Optimizing Immunohistochemistry IHC Assays with Sections of PFPE PAXgene Tissue Fixed Paraffin Embedded Tissue 19 References 20 Ordering Information 21 PreAnalytiX Worldwide 23 PAXgene Tissue Container Product Circular 04 2013 3 Kit Contents PAXgene Tissue Container 10 Catalog no 765112 PAXgene Tissue Container 10 Product circular Contains methanol See page 5 for safety information Symbols Use by Lot number e ra MAT Material number Upper limit of temperature Legal manufacturer E gt Shipping and Storage The PAXgene Tissue Container is shipped at ambient temperature PAXgene Tissue Container can be stored upon receipt at ambient or refrigerated temperatures 2 25 C Do not use containers after their expiration date Intended Use 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 It is the user s responsibility to validate the performance of the PAXgene Tissue Container for any particular use
19. ue samples larger than 4 x 15 x 15 mm as this will significantly reduce the quality of tissue morphology as well as the integrity of nucleic acids Ensure that the container boxes are intact and undamaged and that reagents have not leaked Do not use a container that has been damaged To avoid transterring samples to the wrong container ensure that the containers as well as the tissue cassettes are properly labeled Procedure Excise tissue in preparation for fixation Note Within 30 min of tissue excision sample should be fixed in PAXgene Tissue FIX of chamber 1 If necessary cut tissue to a maximum size of 4 15 15 mm Note Smaller tissue samples or biopsies can be fixed without cutting Place the tissue specimen into a standard tissue cassette not provided Unscrew the cap of the PAXgene Tissue Container and lift up the screw cap rack assembly Attach the tissue cassette to the rack by inserting the lower edge of the tissue cassette into the bottom edge of the rack grasp the cassette so that the slanted angled end faces upwards and toward you and press to secure it under the rack upper release tab see flowchart page 8 Carefully submerge the rack holding the tissue cassette into chamber 1 containing PAXgene Tissue FIX Screw the cap into place Note Make sure that the cap is correctly seated on the rim of the PAXgene Tissue Container before tightening the cap Incubate tissue specimen at room temperature 1
20. xation rates may vary depending upon type and size of tissue 6 PAXgene Tissue Container Product Circular 04 2013 Description of protocols Sample collection and stabilization with the PAXgene Tissue Container PAXgene Tissue Containers are dual chamber containers prefilled with 2 reagents Chamber 1 contains PAXgene Tissue FIX and chamber 2 PAXgene Tissue STABILIZER For fixation tissue samples with a maximum size of 4 15 15 mm must be placed into a standard tissue cassette Fixation is performed in chamber 1 Figure 1 PAXgene Tissue FIX rapidly penetrates and fixes the tissue After fixation the tissue cassette is removed from chamber 1 and transferred to PAXgene Tissue STABILIZER in the second chamber of the same container Figure 2 When the tissue is stored in PAXgene Tissue Stabilizer nucleic acids proteins and morphology of the tissue sample are stable for a minimum of 7 days at room temperature 15 25 C or for a minimum of 4 weeks at 2 8 C depending on the type of tissue Tissue samples can be stored in the PAXgene Tissue STABILIZER for longer periods at 20 to 80 C without negative effects on the morphology of the tissue or the integrity of the nucleic acids 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 Figure 3 See the PAXgene Tissue Kit Handbooks for information about
21. zation within Chamber 2 of the PAXgene Tissue Container Figure 3 Preservation of nucleic acids and morphology Subsequent analysis of fixed and stabilized material shows stable preservation of DNA FN and RNA Microscope examination shows stable preservation of morphology 8 For further information about applications using the PAXgene Tissue System visit www preanalytix com PAXgene Tissue Container Product Circular 04 2013 Equipment and Reagents to Be Supplied by User When working with chemicals always wear a suitable lab coat disposable gloves and protective goggles For more information consult the appropriate satety data sheets SDSs available trom the product supplier e Standard tissue cassette available through EMS VWR Thermo Scientitic and others e Ethanol purity grade a d 96 100 or denatured with methanol isopropanol e g histological grade alcohol composed of 90 parts ethyl alcohol 5 parts methyl alcohol and 5 parts isopropyl alcohol or methyl ethyl ketone i e 99 parts ethanol and 1 part methyl ethyl ketone e Xylene or xylene substitute e Paraffin with a melting point of 54 58 C e g Paraplast X tra Thermo Fisher Scientific cat no 503002 or VWR US cat no 100504 164 e Optional for storage at 20 5 15 C to 30 C or 80 C 65 5 to 90 C cryogenic vial with screw closure e g Thermo Fisher Scientific cat no 5005 0015 e Scalpel e Heated water bath
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