The unique retriever solves the problem of staining formalin fixed tissues. This is a low-cost solution to all known significant difficulties with immunohistochemistry on paraffin slices. The combination of ease of use and great consistency of findings will provide you with the highest quality immunostaining.
The Retriever is a bench-top model for thermally processing slides of formalin-fixed, paraffin embedded tissues prior to immunostaining. The model has been designed to ensure identical processing of all the samples during a processing cycle, as well as the identical processing of the samples in individual sessions. The retriever preserves processed tissues.
Features:
- Run antigen unmasking in 6 various buffers at once.
- Perform gentle antigen retrieval that does not damage the tissue morphology.
- Get identical results every time.
- Compare a series of slides treated in independent sessions.
Full set contains the processing unit, slide chamber (allows to fit 18 Slides) and Chamber Rack for 6 Slide Chambers (108 slides in total).
How does the Retriever work?
Ok, basically it is a pressure cooker. However, a pressure cooker specially designed to unmask the antigen on tissue sections. The core principle is heating of the chambers with the slides at high temperature (>120°C) and high pressure. Sensors control the heating profile for the temperature and
pressure to be reached at certain pace and over certain time. We did a lot of tests to find the optimal settings.
When the required temperature is reached, it will be kept for several minutes. After that the slides will cool over 2 hours. Specially designed thermal walls of the unit control the speed of cooling of the inner chamber and slides.
Who would benefit from using our Retriever?
- Investigative Pathology, where the high quality of staining (a picture may be published!) is required.
- Any lab that is short on technical personnel: any student or post-doc can process slides for the staining without using much time
- Small routine pathology labs, where a limited number of slides should be processed daily
- Anyone who uses highly valuable samples, such as tissue arrays or unique tissue samples: Simplicity and reliability of the unit ensures the safety of your sample, and a high quality antigen unmasking
How To Use:
Step 1.
Deparaffinized sections on slides are placed into the Processing (Tissue Slide) Chambers (EMS62705-01). Retriever can accommodate simultaneously from 1 to 6 Chambers, which allows you to process a series of slides in up to 6 different antigen unmasking buffers within the same session. Fill the chambers with a buffer of your choice.
Step 2.
Place the Chambers into the Rack. Fill the Retriever with 750 ml of deionized/distilled water. Place the Rack into Retriever. Close the lid with a simple twist.
Step 3.
Push the Start button. The tissues will be processed automatically. In about two hours (depends on the load) you can open the lid and proceed with immunostaining.
Technical Specifications of Retriever:
| Height | 335 mm |
|---|---|
| Capacity | 9 litres |
| Max. Instrument length | 228 mm |
| Width | 340 mm |
| Net Weight | 4.5 kilos |
| Internal chamber Dimensions (d/h) | 210/230 mm |
| Max. Load Weight | 3.0kg |
Other Specifications:
Fuses - Located under the control module, fuses F1 OA, 32 x 6.3mm, ceramic sand filled, Mains plug top fuse (User replaceable), F1 3A to BS1362 UK ONLY.
Rating - Models are rated continuously for intermittent use.
Body- Deep drawn aluminium. Lid - Aluminium.
Heater- Externally surface mounted mechanically fixed electric element.
Temperature Cut Out - Thermal fuse.
Pressure - Calibrated pressure release valve.
Max. Single Fault Temperature - 133.3°C
Environment Conditions - indoor use - temperature 5°C to 40°C - altitude up to 2000m - maximum relative humidity 80% for temperatures up to 31°C decreasing linearly to 50% relative humidity at 40°C. - mains supply voltage fluctuations not to exceed +10% of the nominal voltage
Input Connections - Mains inlet socket 'hot' format conforming to IEC 302.
Safety Shut Down - See "Temperature Cut Out".
Choosing A Buffer:
If you already know the buffer that can be used for Microwave treatments of sections in order to unmask you antigen of interest, the high chances are that the same buffer may be used in Retriever.
To improve the morphology of tissue of the processed sections, use one of our supplied ready-to-use buffers by choosing them according to pH.
We have commercially available the following buffers:
- R-Buffer A pH 6.0
- R-Buffer B pH 8.0
- R-Buffer C pH 4.5
- R-Buffer U pH 6.0
- G variants of the same buffers for a more gentle processing of the tissues.
To successfully retrieve the antigen of interest on fixed sections, please remember that two factors define the choice of buffer:
- The nature of the antigen/epitope and of the antibody used for its detection
- The fixative used and the degree of fixation.
Correction for Fixative & Degree of fixation
The suggested protocol for processing tissues is optimized for routinely formalin-fixed and paraffin embedded material. If the tissue used for sections was insufficiently fixed, overfixed (was left in formalin for too long), or other fixative was used, the protocol may require some modifications. The easiest correction is to use EMS's own specially formulated buffers for a gentle (G) tissue processing. Use them instead of the basic buffers. For overfixed material try using U buffer or run the additional cycle in the same buffer.
Fixative Used on Tissues Buffer
| Formalin, buffered | R-Buffer A, B, C or U |
|---|---|
| Formalin-Zn | R-Buffer A, B, C or U |
| Mirsky fixative | Under testing |
| Boonfix I | Under testing |
| Boonfix II | Under testing |
| PLP | R-buffer AG, BG, or CG |
Choosing Buffer: Correction for the Nature of the Antigen and Epitope.
The choice of the buffer depends on the nature of the antigen and the location of the epitope of interest. We advise you to first run the test for the most appropriate buffer using the tissues where the expression of the antigen does occur. General guidelines for choosing the buffer:
Most of the nuclear antigens (apoptosis-related, survival-related, proliferation-related) R: Buffer A (or AG).
Cell adhesion molecules, cell membrane antigens (extracellular domain) R: Buffer A (or AG)
Cytoskeleton and cytoskeleton-associated molecules - R:Buffer A (or AG)
Intracellular domain of some adhesion molecules and surface receptors - R:Buffer B (or BG)
Intracellular domain of some adhesion molecules and surface receptors - R:Buffer C (or CG)
Most of antibodies raised against a linear peptide - R: Buffer U (or UG)
Publications involving examples of the retriever in various types of research and clinical studies:
Cancer Pathology
| Urology. 2006 Sep;68(3):578-82. Inactivation of myopodin expression associated with prostate cancer relapse. Yu YP, Tseng GC, Luo JH.
Acta Oncologica. 2008;47(8):1539-45. Aberrant expression of cyclin E in low-risk node negative breast cancer. Ahlin C, Gruhne B, Holmqvist M, Zetterberg A, Fjällskog ML.
Breast Cancer. 2010 Jul;122(2):315-24. Numb protein expression correlates with a basal-like phenotype and cancer stem cell markers in primary breast cancer. Rennstam K, McMichael N, Berglund P, Honeth G, Hegardt C, Rydén L, Luts L, Bendahl PO, Hedenfalk I.
Pancreas. 2009 Apr;38(3):259-66. Neurogenin 3 and neurogenic differentiation 1 are retained in the cytoplasm of multiple endocrine neoplasia type 1 islet and pancreatic endocrine tumor cells. Lejonklou MH, Edfeldt K, Johansson TA, Stålberg P, Skogseid B.
Hormone and Metabolic Research. 2008 May;40(5):354-9. Lack of nuclear expression of hairy and enhancer of split-1 (HES1) in pancreatic endocrine tumors. Johansson T, Lejonklou MH, Ekeblad S, Stålberg P, Skogseid B.
Leukemia and Lymphoma. 2010 Jul;51(7):1269-77. Potential biomarkers of bortezomib activity in mantle cell lymphoma from the phase 2 PINNACLE trial. Goy A, Bernstein SH, McDonald A, Pickard MD, Shi H, Fleming MD, Bryant B, Trepicchio W, Fisher RI, Boral AL, Mulligan G.
Clinical Cancer Research. 2006 Jul 1;12(13):4080-9. PHA-680632, a novel Aurora kinase inhibitor with potent antitumoral activity. Soncini C, Carpinelli P, Gianellini L, Fancelli D, Vianello P, Rusconi L, Storici P, Zugnoni P, Pesenti E, Croci V, Ceruti R, Giorgini ML, Cappella P, Ballinari D, Sola F, Varasi M, Bravo R, Moll J.
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Cell Biology
| Molecular Biology of the Cell Vol. 20, 2572-2581, May 15, 2009. Regulation of Mammary Gland Branching Morphogenesis by EphA2 Receptor Tyrosine Kinase. David Vaught, Jin Chen, and Dana M. Brantley-Sieders.
Molecular Cancer Therapeutics. 2008 Mar;7(3):590-8. Dual inhibition of VEGFR and EGFR signaling reduces the incidence and size of intestinal adenomas in Apc(Min/+) mice. Alferez D, Wilkinson RW, Watkins J, Poulsom R, Mandir N, Wedge SR, Pyrah IT, Smith NR, Jackson L, Ryan AJ, Goodlad RA.
Molecular Cell BiologyMol Cell Biol. 2004 Apr;24(7):2673-81. Inhibition of Mist1 homodimer formation induces pancreatic acinar-to-ductal metaplasia. Zhu L, Tran T, Rukstalis JM, Sun P, Damsz B, Konieczny SF.
PNAS U S A. 2007 Jun 12;104(24):10063-8. Loss of a quiescent niche but not follicle stem cells in the absence of bone morphogenetic protein signaling. Kobielak K, Stokes N, de la Cruz J, Polak L, Fuchs E.
Science. 2010 Jun 4;328(5983):1278-8. Meiotic recombination provokes functional activation of the p53 regulatory network. Lu WJ, Chapo J, Roig I, Abrams JM.
Cancer Research. 2006 Jan 1;66(1):242-7. Mist1-KrasG12D knock-in mice develop mixed differentiation metastatic exocrine pancreatic carcinoma and hepatocellular carcinoma. Tuveson DA, Zhu L, Gopinathan A, Willis NA, Kachatrian L, Grochow R, Pin CL, Mitin NY, Taparowsky EJ, Gimotty PA, Hruban RH, Jacks T, Konieczny SF.
JCI. 2008 Jan;118(1):89-99. Glypican-1 modulates the angiogenic and metastatic potential of human and mouse cancer cells. Aikawa T, Whipple CA, Lopez ME, Gunn J, Young A, Lander AD, Korc M.
Cancer ResearchCancer Res. 2007 Jan 1;67(1):276-80. Transgenic overexpression of RasGRP1 in mouse epidermis results in spontaneous tumors of the skin. Oki-Idouchi CE, Lorenzo PS.
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Dermatology
| Journal of Investigative Dermatology. (2005) 124, 615-621 Mutations in Gasdermin 3 Cause Aberrant Differentiation of the Hair Follicle and Sebaceous Gland. Declan P Lunny, Erica Weed, Patrick M Nolan, Andreas Marquardt, Martin Augustin and Rebecca M Porter |
Immunology, Research & Clinical
| Journal of Experimental Medicine. 2009 Aug 3;206(8):1661-71. Loss of the lupus autoantigen Ro52/Trim21 induces tissue inflammation and systemic autoimmunity by disregulating the IL-23-Th17 pathway. Espinosa A, Dardalhon V, Brauner S, Ambrosi A, Higgs R, Quintana FJ, Sjöstrand M, Eloranta ML, Ní Gabhann J, Winqvist O, Sundelin B, Jefferies CA, Rozell B, Kuchroo VK, Wahren-Herlenius M.
Bone Marrow Transplantation. 2008 Dec;42(12):807-18.GVHD after chemotherapy conditioning in allogeneic transplanted mice. Sadeghi B, Aghdami N, Hassan Z, Forouzanfar M, Rozell B, Abedi-Valugerdi M, Hassan M.
Journal of Reproductive Immunology. 2010 Mar;84(2):176-85. Pro-inflammatory and anti-inflammatory cytokines in human preterm and term cervical ripening. Dubicke A, Fransson E, Centini G, Andersson E, Byström B, Malmström A, Petraglia F, Sverremark-Ekström E, Ekman-Ordeberg G.
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Neuroscience
| Journal of Comparitive Neurology. 2008 Sep 1;510(1):79-99. Distinct molecular pathways for development of telencephalic interneuron subtypes revealed through analysis of Lhx6 mutants. Zhao Y, Flandin P, Long JE, Cuesta MD, Westphal H, Rubenstein JL. |
Reproductive Biology
| Fertility & Sterility. Vol 91, Issue 4, Supplement, Pages 1420-1423 (April 2009) Expressions of steroid receptors and Ki67 in first-trimester decidua and chorionic villi exposed to levonorgestrel used for emergency contraception Chun-Xia Meng, M.D.ab, Li-Nan Cheng, M.D.b, Parameswaran Grace Luther Lalitkumar, Ph.D.a, Lin Zhang, M.D.b, Hui-Juan Zhang, M.D.b, Kristina Gemzell-Danielsson, M.D., Ph.D.
Human Reproduction. 2010 Apr;25(4):874-83. Effects of oral and vaginal administration of levonorgestrel emergency contraception on markers of endometrial receptivity. Meng CX, Marions L, Byström B, Gemzell-Danielsson K.
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Virology. 2009 Oct 25;393(2):338-45. An early 'classical' swine H1N1 influenza virus shows similar pathogenicity to the 1918 pandemic virus in ferrets and mice. Memoli MJ, Tumpey TM, Jagger BW, Dugan VG, Sheng ZM, Qi L, Kash JC, Taubenberger JK.
mBio 2010 1(1):e00067-10. doi:10.1128 The PB2-E627K mutation attenuates viruses containing the 2009 H1N1 influenza pandemic polymerase. Jagger, B. W., M. J. Memoli, Z.-M. Sheng, L. Qi, R. J. Hrabal, et al.
OnePLoS One. 2009 Jun 16;4(6):e5916. Infectious prions in pre-clinical deer and transmission of chronic wasting disease solely by environmental exposure. Mathiason CK, Hays SA, Powers J, Hayes-Klug J, Langenberg J, Dahmes SJ, Osborn DA, Miller KV, Warren RJ, Mason GL, Hoover EA.
Journal of Virology. 2009 Apr;83(8):3754-61. Role of sialic acid binding specificity of the 1918 influenza virus hemagglutinin protein in virulence and pathogenesis for mice. Qi L, Kash JC, Dugan VG, Wang R, Jin G, Cunningham RE, Taubenberger JK.
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