EnzMet:
for in situ hybridization, immunohistochemistry,
electron microscopy, correlative microscopy, and biochips

See what you've been missing
in immunohistochemistry (IHC)
and in situ hybridization (ISH)

• What is EnzMet?
  
• Features of EnzMet
• Applications
  • In situ hybridization (ISH)
  • Immunohistochemistry (IHC)
  • Electron Microscopy
  • Light and Electron Correlative Microscopy
  • Conductive Array Biochip Fabrication
  • Enzmet is saving lives: Her2 breast cancer test
• Ordering and Product Information
• Instructions, Methods and Protocols
• Tips and Tricks
• References
• Other Nanoprobes in situ hybridization technologies

What is EnzMet?
Clearer, crisp staining for
in situ hybridization (ISH)
and immunohistochemistry (IHC)

EnzMet (Enzyme Metallography) is a new biological labeling and staining method developed at Nanoprobes. It uses a targeted enzymatic probe with a novel metallographic substrate to provide a quantum leap in staining clarity over conventional chromogenic and fluorescent substrates.

EnzMet™ 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.

 

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• 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.

EnzMet™'s 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 microscopy method.

Human breast cancer biopsy tissue section where single copies (2 per normal cell) of the Her 2/neu gene were detected by EnzMet (black spots).

Better than FISH:
EnzMet enables bright field detection, a permanent signal, and use of full-strength H&E staining for simultaneous visualization of underlying tissue morphology.

Original magnification x 400; courtesy of Dr. Raymond R. Tubbs, Cleveland Clinic Foundation.

• In situ hybridization (ISH)
  [Feature article]
  Highly sensitive in situ hybridization, with single-copy sensitivity and copy counting.^1,2
  

• Immunohistochemistry (IHC)
  [Feature article]
  Precise, sensitive and clear immunohistochemical staining and detection.³
  

• HER2 staining
  [Feature article]
  Simultaneous ISH/IHC assay for HER2 gene amplification and concomitant oncoprotein overexpression (using Fast Red K IHC with EnzMet ISH).⁴
  

• Biochip Fabrication
  [Feature article]
  Highly specific electrical detection using conductive array biochips: enzyme-labeled DNA probes were developed with EnzMet to form conductive bridges.⁵
  

• Correlative microscopy
  [Feature article]
  Correlative light and electron microscopic localization of microsporidial spore proteins.⁶
  

• Complete references for EnzMet
• EnzMet references, sorted by application

• Immonodot and Protein Blots
  Highly sensitive visualization on immunodot blots and other protein blots with negligible background.