A new class of electron microscopy labels is being developed that has 5-10 times better resolution than those currently available. This class promises to be the next generation of high resolution electron microscopic labels and should have an extensive impact on molecular biology, since it should make possible direct domainal mapping of single biomolecules and macromolecular assemblies. Previously we were able to introduce undecagold (Au11) as a specifically bound protein label. We also functionalized it in several ways to demonstrate its utility in labeling sulfhydryls, amino groups and carbohydrate residues in glycoproteins. Pioneering work in this new field will continue along 3 lines: 1) Synthetic chemistry to produce a wider range of labels with varying properties and reactive groups. 2) Basic physical studies to properly characterize these labels covering their visibility, detection limits and sensitivity to the electron beam. 3) Biological applications to demonstrate the variety of labeling available, its reliability, as well as answering some important biological structural questions not easily possible with any other techniques. Chemical synthesis and characterization will focus on development of a monofunctional undecatungsten (W11) label that has just recently been successfully made for the first time. This W11 label (with organic side chain) has several advantages over the Au11 label and will be derivatized to react with various protein, nucleic acid and carbohydrate moieties. Biological applications will include labeling of: t-RNA and ribosome components, mutienzyme complexes (pyruvate dehydrogenase complex), actin, myosin, intermediate filaments, human fibrinogen, acetytcholine receptors and membranes. Fab fragments will also be labeled to provide an antibody label 5 times smaller than those currently available. A new electron gun allowing low dose/high dose data to be collected with no optical changes will enable minimally damaged specimens to be used. A stable cold stage will permit frozen hydrated cluster-labeled samples to be observed. Image processing to optimally recover high resolution data will also be employed especially in radial and helical reconstructions. We will draw upon our large user program of the STEM NIH Biotechnology Resource to provide interesting samples and biochemical expertise since many users are very anxious to try these new labels.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
Project #
Application #
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Associated University-Brookhaven National Lab
United States
Zip Code
Hainfeld, J F; Furuya, F R (1992) A 1.4-nm gold cluster covalently attached to antibodies improves immunolabeling. J Histochem Cytochem 40:177-84
Watts, N R; Hainfeld, J; Coombs, D H (1990) Localization of the proteins gp7, gp8 and gp10 in the bacteriophage T4 baseplate with colloidal gold: F(ab)2 and undecagold: Fab' conjugates. J Mol Biol 216:315-25
Hainfeld, J F; Foley, C J; Maelia, L E et al. (1990) Eleven tungsten atom cluster labels: high-resolution, site-specific probes for electron microscopy. J Histochem Cytochem 38:1787-93
Hainfeld, J F; Lipka, J J; Quaite, F E (1990) A high-resolution tungstate membrane label. J Histochem Cytochem 38:1795-803
Hainfeld, J F; Foley, C J; Srivastava, S C et al. (1990) Radioactive gold cluster immunoconjugates: potential agents for cancer therapy. Int J Rad Appl Instrum B 17:287-94
Hainfeld, J F (1987) A small gold-conjugated antibody label: improved resolution for electron microscopy. Science 236:450-3
Monson, K L; Wall, J S; Hainfeld, J F (1987) Visibility and stability of a 12-tungsten atom complex in the scanning transmission electron microscope. Ultramicroscopy 21:147-56
Wall, J S; Hainfeld, J F (1986) Mass mapping with the scanning transmission electron microscope. Annu Rev Biophys Biophys Chem 15:355-76
Hainfeld, J F; Wall, J S (1986) Mapping the domains of molecules and complexes by mass and heavy atom loading. Ann N Y Acad Sci 483:181-7