Preparation, use, and enlargement of ultrasmall gold particles in immunoelectron microscopy.
The introduction of ultrasmall (approximately 1-3 nm) colloidal gold markers in immunoelectron microscopy (IEM) in 1989 has considerably improved the sensitivity of this marker system. Ultrasmall gold markers have opened the field of pre-embedding labeling studies to gold markers without the need of harsh permeabilizing steps. They are recommended for the detection of scarce antigens in ultrathin cryosections which may otherwise escape immunodetection. However, reports concerning the preparation of ultrasmall gold colloids, their conjugation to proteins, and their use in high-resolution studies (without an additional enlargement step) are very limited. Also, the available enlargement techniques necessary for the use of this marker in conventional electron microscopy require detailed discussion to clarify the large number of contradictory observations. The present review summarizes and discusses the findings accumulated within the last 10 years on the application of ultrasmall gold markers in IEM with regard to their merits, limitations, detection sensitivity, and suitability for different labeling techniques. It should provide practical hints for the use of ultrasmall gold colloids and discusses problems arising with enlargement techniques such as silver enhancement and gold toning procedures.
Maurice E. Mueller Institute for Structural Biology at the Biozentrum, University of Basel, Switzerland.
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Baschong
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Stierhof
D007089:Image Enhancement; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron / Q000379:methods*