EnzMet™: See what you’ve been missing

Updated: March 13, 2008 Contents What is EnzMet™? Clearer, crisp staining for IHC and ISH Features of EnzMet™ Applications Product and Catalog Information References What is EnzMet™? Clearer, crisp staining for IHC and ISH EnzMet™ (Enzyme Metallography) is a new biological labeling and staining method developed at Nanoprobes, which uses a targeted enzymatic probe with a novel metallographic substrate to provide a quantum leap in staining clarity over conventional chromogenic and fluorescent substrates. It has proven highly sensitive both for in situ hybridization (ISH), where it readily visualizes endogenous copies of single genes, and immunohistochemistry (IHC) detection. It has also been used as an electrical detection method for biochips.

EnzMet™ immunohistochemistry. (left) Mechanism of the enzyme metallographic process showing enzyme-catalyzed deposition of metal from solution. (right) Paraffin-embedded Human bladder adenocarcinoma immunostained for cytokeratin with immunoperoxidase with DAB vs. EnzMet.

Features of EnzMet™

Patented EnzMet™ technology uses HRP to deposit metallic silver with extraordinary selectivity. High sensitivity: detect single copies of target genes, or low-abundance proteins with virtually no background. Black, sharply defined, non-diffusing stain gives higher resolution localization Compatible with all counterstains: lets you clearly see surrounding tissue morphology (as opposed to fluorescent signals where surrounding tissue is not visible) Minimal diffusion: super-high resolution compared with DAB. Near zero background. Does not fade or bleach.

The sharply resolved black signal is readily distinguished from other stains: it has been combined with fast red K immunohistochemistry to provide a concomitant brightfield gene and protein assay. EnzMet™ has many advantages both over fluorescent labels and enzyme chromogens. Because it is used in the conventional brightfield microscope, it does not require fluorescent optics, or dark adaptation on the part of the user. The signal is permanent, and does not have the photobleaching problems associated with fluorescent stains. The high density enables visualization at even low magnification. It produces a sharp metal deposit that does not diffuse like DAB, giving higher resolution localizations. Because the deposited metal is electron-dense, it provides high contrast for electron microscopy, making enzyme metallography a potential correlative light and electron microscopic method.

Nanoprobes recently teamed with Ventana Medical Systems, Incorporated, for the commercial development and use of this reagent in automated slide staining instruments and applications. Introduction of SISH (Silver In Situ Hybridization) for detecting Her 2 breast cancer based on EnzMet™ has now been released in Europe and scheduled for U.S. use is pending FDA approval. Please contact Ventana Medical Systems for all automated slide staining applications.

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Applications

Complete references for EnzMet™
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Catalog and Product Information

6001	EnzMet HRP Detection Kit for IHC/ISH * Enzyme Metallographic substrate for immunohistochemical and in situ hybridization applications. 30 mL total, enough for 150 slides. Solution A / Solution B / Solution C 18 mL / 6 mL / 6 mL Product information and instructions (HTML)   Product information (PDF)	$93
6002	EnzMet Western Blot HRP Detection Kit Enzyme Metallographic substrate for blot applications. 100 mL, sufficient for 15 blots. Solution A / Solution B / Solution C 60 mL / 20 mL / 20 mL Product information and instructions (HTML)   Product information (PDF)	$248
6002	EnzMet HRP Detection Kit for Research Applications Enzyme Metallographic substrate for general research applications. 45 mL total. Solution A / Solution B / Solution C 15 mL / 15 mL / 15 mL Product information and instructions (HTML)   Product information (PDF)	$230
	* Not approved for clinical use. Contact Ventana Medical Systems for clinical applications.

To place an order, you can Order directly with our online order form, or contact us:

References

1. Powell, R. D.; Pettay, J. D.; Powell, W. C.; Roche, P. C.; Grogan, T. M.; Hainfeld, J. F., and Tubbs, R. R.: Metallographic in situ hybridization. Hum. Pathol., 38, 1145-1159 (2007).
   • Abstract (courtesy of Science Direct).

2. Tubbs, R.; Pettay, J.; Hicks, D.; Skacel, M.; Powell, R.; Grogan, T., and Hainfeld, J.: Novel bright field molecular morphology methods for detection of HER2 gene amplification. J. Mol. Histol., 35, 589–594 (2004).
   • Abstract (courtesy of the SpringerLink).

3. Tubbs R.; Pettay J.; Powell R.; Hicks D. G.; Roche P.; Powell W.; Grogan T., and Hainfeld, J. F.: High-resolution immunophenotyping of subcellular compartments in tissue microarrays by enzyme metallography. Appl. Immunohistochem. Mol. Morphol., 13, 371-375 (2005).
   • Abstract (courtesy of the PubMed (Medline)).

4. Downs-Kelly, E.; Pettay, J.; Hicks, D.; Skacel, M.; Yoder, B.; Rybicki, L.; Myles, J.; Sreenan, J.; Roche, P.; Powell, R.; Hainfeld, J.; Grogan, T., and Tubbs, R.: Analytical Validation and Interobserver Reproducibility of EnzMet GenePro: A Second-Generation Bright-Field Metallography Assay for Concomitant Detection of HER2 Gene Status and Protein Expression in Invasive Carcinoma of the Breast. Am. J. Surg. Pathol., 29, 1505-1511 (2005).
   • Abstract (courtesy of SpringerLink).

5. Moller, R.; Powell, R. D.; Hainfeld, J. F., and Fritzsche, W.: Enzymatic control of metal deposition as key step for a low-background electrical detection for DNA chips. Nano Lett., 5, 1475-1482 (2005).
   • Abstract (courtesy of ACS Publications).

6. Powell, R.; Joshi, V.; Thelian, A.; Liu, W.; Takvorian, P.; Cali, A., and Hainfeld, J.: Light and Electron Microscopy of Microsporida using Enzyme Metallography. Microsc. Microanal., 12, (Suppl. 2: Proceedings); Kotula, P.; Marko, M.; Scott, J.-H.; Gauvin, R.; Beniac, D.; Lucas, G.; McKernan, S., and Shields, J. (Eds.), Cambridge University Press, New York, NY, 424CD (2006).
   • Complete paper from Microscopy & Microanalysis

Complete references for EnzMet™
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