Our Unique Retriever solves the problem of staining formalin fixed tissues. This is an affordable solution to all known major problems with immunohistochemistry on paraffin sections. Ease of use combined with high reproducibility of the results will give you the best 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.
Now you can:
- 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 series of slides treated in independent sessions.
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 be cooling 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 labs thatis 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:
Deparaffinized sections on slides are placed into the Processing (Tissue Slide) Chambers. 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.
Place the Chambers into the Rack. Fill the Retriever with 750 ml of deionized/distilled water. Place the Rack into Retriever. Close the lid by a simple twist.
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 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
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.
Examples of Staining :
Ki-67 (Nuclear antigen).
Sections of formalin-fixed, paraffin embedded tissue of human sigmoid were deparaffinized and processed in Retriever to unmask the antigen. We used one cycle in R-Buffer C (pH 6.0) with cooling of the slides overnight. Antibody MIB-1 against proliferation marker Ki-67 (nuclear) was used together with R-Detect HRP detection system for immunostaining of sections.
PCNA (Nuclear antigen).
Sections of formalin-fixed, paraffin embedded tissue of human duodenum were deparaffinized and processed in Retriever to unmask the antigen. We used one cycle in R-Buffer C (pH 6.0) with cooling of the slides overnight. Antibody PC-10 against proliferation marker PCNA nuclear) was used together with R-Detect HRP detection system for immunostaining of sections.
Sections of formalin-fixed, paraffin embedded tissue of human sigmoid were deparaffinized and processed in Retriever to unmask the antigen. We used one cycle in R-Buffer A (pH 8.1) with cooling of the slides overnight. Antibody #4B11 against T-cell marker CD8 (cell membrane) was used together with R-Detect HRP detection system for immunostaining of sections.
E-cadherin (Membranous, extracellular domain).
Sections of formalin-fixed, paraffin embedded tissue of human cervix were deparaffinized and processed in Retriever to unmask the antigen. We used one cycle in R-Buffer C (pH 6.0) with cooling of the slides overnight. Antibody HECD-1 against cell adhesion molecule E-cadherin was used together with R-Detect HRP detection system for immunostaining of sections.
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)
The operation manual for the Retriever is online. Please click here.
Publications: Real-world examples of the retriever in various types of research and clinical studies.
- Cancer Pathology
- Cell Biology
- Immunology, Research & Clinical
- Reproductive Biology
- Virology & Microbiology
- 3 per pack