PAXgene Tissue FIX Container (50 ml)
Contents
1. Sample collection and stabilization with the PAXgene Tissue FIX Container PAXgene Tissue FIX Containers are single chamber containers prefilled with 50 ml of PAXgene Tissue FIX PAXgene Tissue FIX Containers can accommodate four 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 PAXgene Tissue FIX is removed and replaced by PAXgene Tissue STABILIZER within the same container When fixed tissue is stored in PAXgene Tissue STABILIZER the nucleic acids proteins and morphology of the tissue sample are stable for up to 7 days at room temperature 15 25 C or for up to 4 weeks at 2 8 C depending on tissue type 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 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 See the PAXgene Tissue Container Product Circulars for information about DNA RNA or miRNA isolation and the PAXgene Tissue supplementary protocols2. 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 800 789 544 Fax 02 334304 826 Technical 800 787980 Japan Telephone 03 6890 7300 Fax 03 5547 0818 Technical 03 6890 7300 Korea South Orders 080 000 7146 Fax 02 2626 5703 Technical 080 000 7145 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 436 The Netherlands Orders 0800 0229592 Fax 0800 0229593 Technical 0800 0229602 Norway Orders 800 18859 Fax 800 18817 Technical 800 18712 Singapore Orders 1800 742 4362 Fax 65 6854 8184 Technical 1800 742 4368 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 qiagen 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 Fox 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 4
3. Embedding and Sectioning Important points before starting e Samples for processing must first be incubated in PAXgene Tissue FIX for an appropriate length of time see Step 4 on page 13 and then incubated in PAXgene Tissue STABILIZER for a minimum of 2 h e Refer to Appendices A and B for detailed processing protocols pages 16 18 e Do not begin tissue processing either with 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 99 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 Things to do before starting e Cut larger samples to a size that will fit in a standard tissue cassette e Dilute PAXgene Tissue STABILIZER concentrate with ethanol as indicated on the bottle Procedure 1 Transfer the tissue cassette into the first station of a tissue processor filled with 80 99 ethanol to start dehydration 2 Follow a protoco
4. 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 C 15 C to 30 C or 80 C 65 C to 90 C cryogenic vial with screw closure e g Thermo Fisher Scientific cat no 5005 0015 e Scalpel e Heated water bath This is not a complete list of suppliers and does not include many important vendors of biological supplies 10 PAXgene Tissue FIX Container 50 ml Product Circular 02 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 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 paraffin embedded tissue store PFPE blocks at 20 C 15 C to 30 C Labeling of PAXgene Tissue FIX Container In order to allow calculation of the exact length of fixation time record the date and time that tissue is placed in the PAXgene Tissue FIX Container in the Fixation Date Time field on the Container label Figure 4 After replacing PAXgene Tissue FIX with PAXgene Tissue
5. www preanalytix com for protein purification and other applications See Figure 1 on page 8 for an illustration of the steps in the fixation process See Figure 2 on page 9 for an illustration of the steps in the stabilization process Fixation rates and stabilization times depend on type and size of tissue Specifications for tissue size fixation and storage conditions using PAXgene Tissue FIX and PAXgene Tissue STABILIZER were determined using animal tissues samples For the latest results on long term storage see Technical Notes at www preanalytix com PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 7 Multiple smaller tissue samples o M lt 4x15x15 mm ot 0 6 3 Single larger tissue sample C 20 x 20 x 20 mm Resect and cut the tissue into max 4 x 15x 15 mm sections Place in up to four 4 tissue cassettes Place tissue cassettes in PAXgene Tissue FIX Container containing PAXgene Tissue FIX reagent Fixation for 2 to 24 hours depending on tissue sample and size Tissue sample can have max dimensions 20 x 20 x 20 mm Place tissue directly into PAXgene Tissue FIX Container containing PAXgene Tissue FIX reagent Fixation for 6 to 48 hours depending on tissue type and size Figure 1 Fixation Resection and fixation of tissue samples using the PAXgene Tissue FIX Container is a straightforward two step process It is suitable for multiple smaller tissue samples FN or a
6. 0 60 min 18 22 C 0 5 bar 10 Xylene 30 60 min 18 22 C 0 5 bar 11 Paraplast X tra 30 60 min 56 C 0 5 bar 12 Paraplast X tra 90 min 56 C 0 5 bar Optimal incubation times depend on the thickness of the tissue specimen PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 17 Table 3 Processing protocol B Step Media Time Temperature Vacuum 80 Ethanol 15 min 12h 15 25 C 2 90 Ethanol 30 min 15 25 C 0 5 bar 3 95 Ethanol 30 60 min 15 25 C 0 5 bar 4 99 Ethanol 30 60 min 15 25 C 0 5 bar 5 99 Ethanol 30 60 min 15 25 C 0 5 bar 6 Isopropanol 30 60 min 15 25 C 0 5 bar 7 Isopropanol 30 60 min 15 25 C 0 5 bar 8 Xylene 30 60 min 15 25 C 0 5 bar 9 Xylene 30 60 min 15 25 C 0 5 bar 10 1 1 mixture of 30 60 min 50 C 0 5 bar Paraplast X tra and xylene 1 Paraplast X tra 30 60 min 56 C 0 5 bar 12 Paraplast X tra 60 min 56 C 0 5 bar Optimal incubation times depend on the thickness of the tissue specimen 18 PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 Appendix C 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 by heating or proteolytic digestion However it is important to note t
7. 5 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 Orders 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 FIX Container 50 ml Product Circular 02 2013 23 PreAnalytiX A QIAGEN BD Company 02 2013
8. None mandatory 5 99 Ethanol None mandatory 6 99 Ethanol Isopropanol 7 99 Ethanol Isopropanol 8 Xylene Xylene substitutes may be used but do not use clearing agents based on D limonene 9 Xylene Xylene substitutes may be used but do not use clearing agents based on D limonene 10 Paraffin with melting 1 1 mixture of paraffin and xylene point 54 58 C 11 Paraffin with melting None mandatory point 54 58 C 12 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 Contaminations in the alcohol used for sample processing can lead to significant reductions in DNA and RNA yield Use filtered or deionized water for preparation of 80 90 or 9596 ethanol PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 Appendix B Other Processing Protocols Successfully Tested for Use with Specimens Treated with the PAXgene Tissue System Table 2 Processing protocol A Step Media Time Temperature Vacuum PAXgene Tissue up to 7 days 18 22 C STABILIZER 2 99 Ethanol 30 60 min 18 22 C 0 5 bar 3 99 Ethanol 30 60 min 18 22 C 0 5 bar 4 99 Ethanol 30 60 min 18 22 C 0 5 bar 5 99 Ethanol 30 60 min 18 22 C 0 5 bar 6 9996 Ethanol 30 60 min 18 22 C 0 5 bar 7 Isopropanol 30 60 min 18 22 C 0 5 bar 8 Isopropanol 30 60 min 18 22 C 0 5 bar 9 Xylene 3
9. PAXgene Tissue FIX Container 50 ml Product Circular For fixation and stabilization of tissue specimens Important To be used in conjunction with PAXgene Tissue STABILIZER For research use only Not for use in diagnostic procedures e PreAnalytiX L A QIAGEN BD Company February 2013 Trademarks PAXgene PreAnalytiX PreAnalytiX GmbH QIAGEN MinElute Qproteome QIAGEN Group Paraplast Paraplast X tra Thermo Fisher Scientific Thermo Fisher Scientific or its subsidiaries Limited License Agreement for the PAXgene Tissue FIX Container Use of this product signifies the agreement of any purchaser or user of the product to the following terms 1 The product may be used solely in accordance with the protocols provided with the product and this product circular and for use with components contained in the kit only PreAnalytiX grants no license under any of its intellectual property to use or incorporate the enclosed components of this kit with any components not included within this Kit except as described in the protocols provided with the product this product circular and additional protocols available at www preanalytix com Other than expressly stated licenses PreAnalytiX makes no warranty that this kit and or its use s do not infringe the rights of third parties This kit and its components are licensed for one time use and may not be reused refurbished or resold PreAnalytiX specifically disclai
10. STABILIZER record the date and time of the exchange in the Exchange with Tissue STABILIZER Date Time field and mark the checkbox beside the field as shown in Figure 4 PAXgene Tissue FIX Container 50 ml e agg cm KM pUE Tum MEC MENOR For Research Use Only Not for Use in Diagnostic Procedures Identification Number sss Patient DOB a Person Collecting Physician Fixation DateTime 25 C A WO s Se ee ee ee ee wil PreAnalytiX GmbH 2c Exchange with Tissue STABILIZER Date Time 8634 Hombrechtikon CH Figure 4 Label of PAXgene Tissue FIX Container PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 11 Protocol Sample Fixation and Stabilization Starting material Starting material can be up to four tissue samples with a maximum size of 4 x 15 x15 mm placed into standard tissue cassettes or a single tissue sample with a maximum size of 20 x 20 x 20 mm Important points before starting e Do not use tissue samples larger than 20 x 20 x 20 mm as this will significantly reduce the quality of tissue morphology and the integrity of nucleic acids e Do not reuse PAXgene Tissue FIX Containers as this will significantly reduce the quality of tissue morphology and the integrity of nucleic acids e PAXgene Tissue STABILIZER is not supplied with the PAXgene Tissue FIX Container and must be ordered separately cat no 765512 e Ensure that the container boxes are intact and undamaged and that rea
11. ance of the PAXgene Tissue FIX Container for any particular use since the performance characteristics of these Containers have not been validated for any specific organism The performance characteristics of this product have not been fully established 4 PAXgene Tissue FIX Container 50 ml Product Circular 02 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 safety data sheets SDSs These are available online in convenient and compact PDF format at www giagen com Safety where you can find view and print the SDS for each PreAnalytiX kit and kit component PAXgene Tissue FIX Container 50 ml Contain methanol and acetic acid Highly Flammable Toxic Risk and safety phrases R11 23 24 25 36 38 39 23 24 25 S7 16 36 37 39 45 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 PAXgene Tissue FIX Container is tested against predetermined specifications to ensure consistent product quality R11 Highly flammable R23 24 25 Toxic by inhalation in contact with skin and if swallowed R36 38 Irritating to eyes and skin R39 23 24 25 Toxic danger of very ser
12. cation 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 min The reagent preserves morphology and biomolecules without the destructive cross linking and degradation found with formalin fixed tissues The 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 total RNA RNA including miRNA or DNA from PAXgene Tissue fixed and stabilized tissue samples require 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 pre filled 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 analysis Tissue penetration and fixation rates may vary depending upon tissue type and size 6 PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 Description of protocols
13. e Tissue DNA For 50 DNA preps PAXgene DNA Mini 767134 Kit 50 Spin Columns Processing Tubes Microcentrifuge Tubes Carrier RNA and Buffers to be used in conjunction with PAXgene Tissue Containers For up to date licensing information and product specific disclaimers see the respective PreAnalytiX or QIAGEN kit handbook 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 FIX Container 50 ml Product Circular 02 2013 PreAnalytiX Worldwide PreAnalytiX products are distributed by QIAGEN and BD companies Australia Orders 1 800 243 800 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 86 21 3865 3865 Fax 86 21 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
14. gents have not leaked Do not use a container that has been damaged e To avoid transferring samples to the wrong container ensure that the containers and the tissue cassettes are properly labeled Things to do before starting e Dilute PAXgene Tissue STABILIZER concentrate with ethanol as indicated on the bottle Procedure 1 Resect tissue in preparation for fixation Note After tissue resection sample should be fixed in PAXgene Tissue FIX within 30 min 2 Cut tissue into desired sample size based on guidelines under Starting material above Important 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 permeability of the fixative reagent 12 PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 3 If using smaller samples place in standard tissue cassettes and place these into the prefilled PAXgene Tissue FIX Container such that each cassette is completely submerged If using a larger sample place it directly into the prefilled PAXgene Tissue FIX Container such that it is completely submerged Note If liquid overflows the edge of the container sample submersion this is an indication that the tissue specimen s were too large Multiple containers should be used instead Incubate tissue specimen s at room temperature 15 25 C for a minimum of 2 hours but preferab
15. hat since many antibodies used in immunohistochemical assays were developed for use with formalin fixed tissue it is offen necessary to optimize antigen retrieval steps and or to adjust antibody concentration in PFPE tissue in order to achieve optimal staining intensities For the latest information on IHC protocols and staining conditions see the tissue atlas at http www preanalytix com product catalog tissue tissue atlas PAXgene Tissue FIX Container 50 ml Product Circular 02 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 FIX Container 50 ml Product Circular 02 2013 Ordering Information Product Contents PAXgene Tissue FIX For fixation and stabilization of tissue Container 50 ml specimens 10 Prefilled Reagent Containers containing 50 ml of PAXgene Tissue FIX Related products PAXgene Tissue STABILIZER for stabilization of tissue samples treated with PAXgene Tissue PAXgene Tissue 8 bottles of PAXgene Tissue STABILIZER STABILIZER Concentrate co
16. ious irreversible effects through inhalation in contact with skin and if swallowed S7 Keep container tightly closed S16 Keep away from sources of ignition No smoking S36 37 39 Wear suitable protective clothing gloves and eye face protection S45 In case of accident or if you feel unwell seek medical advice immediately show the label where possible PAXgene Tissue FIX Container 50 ml Product Circular 02 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 In order 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 pre filled containers for tissue collection storage and transportation and kits for purification of RNA DNA or total RNA including miRNA In addition supplementary protocols for protein purifi
17. l for paraffin embedding See Appendix A on page 16 for a generic protocol or Appendix B on page 17 for examples of two 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 14 PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 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 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 FIX Container 50 ml Product Circular 02 2013 15 Appendix A Generic Processing Protocol That Supports Preservation of Biomolecules in Specimens Treated with the PAXgene Tissue System Table 1 Generic processing protocol Step Media Alternatives 80 99 Ethanol PAXgene Tissue STABILZER 2 90 99 Ethanol None mandatory 3 95 99 Ethanol None mandatory 4 99 Ethanol
18. ly 3 24 hours for samples up to 4x15x 15 mm or 6 48 hours but preferably 8 24 hours for samples up to 20 x 20 x 20 mm Note Longer fixation periods than indicated for the sample sizes 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 Discard PAXgene Tissue FIX and replace with PAXgene Tissue STABILIZER Note A minimum incubation time of 2 h in PAXgene Tissue STABILIZER is recommended before processing and embedding in paraffin Note Instead of exchanging PAXgene Tissue FIX with PAXgene Tissue STABILIZER tissue can be removed from the container and transferred to a tissue processor filled with PAXgene Tissue STABILIZER at position one See Table 2 Appendix B on page 16 for an example of a processing protocol using PAXgene Tissue STABILIZER Process the samples further or store or transport tissue sample in PAXgene Tissue STABILIZER Note Standard storage conditions are up to 7 days at room temperature 15 25 C 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 80 C 65 C to 90 C For freezing we recommend transferring the tissue sample from the PAXgene Tissue FIX Container into a screw cap cryogenic vial filled with PAXgene Tissue STABILIZER PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 13 Protocol Sample Processing Paraffin
19. ms any other licenses expressed or implied other than those expressly stated Uy es 400r fh The purchaser and user of the kit agree not to take or permit 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 Kit and or its components For updated license terms see www preanalytix com 2013 PreAnalytiX GmbH all rights reserved PreAnalytiX 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 at PreAnalytiX GmbH Please see the last page for contact information for your local PreAnalytiX distributor Contents Kit Contents Symbols Shipping and Storage Intended Use Safety Information Quality Control Introduction Principle and procedure Description of protocols Equipment and Reagents to Be Supplied by User ON 0o O0 UuUu n n A Important Notes Storage and archiving of PFPE PAXgene Tissue fixed paraffin embedded tissue samples Labeling of PAXgene Tiss
20. ncentrate for 4 liters of PAXgene 150 ml Tissue STABILIZER PAXgene Tissue Containers for collection fixation and nucleic acid stabilization of human tissues PAXgene Tissue For collection fixation and stabilization Containers 10 of 10 samples 10 Prefilled Reagent Containers containing PAXgene Tissue Fix and PAXgene 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 Microcentrifuge Tubes Carrier RNA RNase Free DNase and RNase Free Buffers to be used in conjunction with PAXgene Tissue Containers PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 Cat no 765312 765512 765112 765134 21 Product Contents Cat no 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 766134 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 DNA Kit for purification of DNA from tissues samples fixed and stabilized using the PAXgene Tissue System PAXgen
21. single larger tissue sample f 8 PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 Pour off PAXgene Tissue FIX Fill with PAXgene Tissue STABILIZER After processing and paraffin embedding block of PFPE PAXgene Tissue fixed paraffin embedded tissue ready for sectioning Figure 2 Stabilization Tissue stabilization involves a single reagent change Stabilized tissues can be stored immediately used for nucleic acid and protein analysis or as illustrated processed and paraffin embedded for sectioning A Figure 3 Preservation of nucleic acids and morphology Later analysis of fixed and stabilized material shows stable preservation of DNA M and RNA Microscope examination shows stable preservation of morphology PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 9 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 safety data sheets SDSs available from the product supplier e PAXgene Tissue STABILIZER Concentrate 150 ml PreAnalytiX cat no 765512 e Standard tissue cassette available from EMS VWR Thermo Scientific and others e Ethanol purity grade a d 96 10096 or denatured with methanol isopropanol e g histological grade alcohol composed of 90 parts ethyl alcohol 5 parts methyl alcohol and 5 parts isopropyl
22. ue FIX Container Protocol Sample Fixation and Stabilization 12 Protocol Sample Processing Paraffin Embedding and Sectioning 14 Appendix A Generic Processing Protocol That Supports Preservation of Biomolecules in Specimens Treated with the PAXgene Tissue System16 Appendix B Other Processing Protocols Successfully Tested for Use with 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 Ordering Information 21 PreAnalytiX Worldwide 23 PAXgene Tissue FIX Container 50 ml Product Circular 02 2013 3 Kit Contents PAXgene Tissue FIX Container 50 ml 10 Catalog no 765312 PAXgene Tissue FIX Container 50 ml 10 Product Circular Contains methanol See page 5 for safety information Symbols Use by Lot number Material number Upper limit of temperature Legal manufacturer Bs g Shipping and Storage The PAXgene Tissue FIX Container is shipped at ambient temperature The PAXgene Tissue FIX Container can be stored at room temperature or refrigerated temperature 2 25 C An expiration date has not been determined 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 perform
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