On the experimental use of light metal salts for negative staining.
All common negative stains are salts of heavy metals. To remedy several technical defects inherent in the use of heavy metal compounds, this study investigates whether salts of the light metals sodium, magnesium, and aluminum can function as negative stains. Screening criteria require aqueous solubility at pH 7.0, formation of a smooth amorphous layer upon drying, and transmission electron microscope imaging of the 87-A (8.7-nm) lattice periodicity in thin catalase crystals. Six of 23 salts evaluated pass all three screens; detection of the protein shell in ferritin macromolecules indicates that light metal salts also provide negative staining of single particle specimens. Appositional contrast is less than that given by heavy metal negative stains; image density can be raised by increasing electron phase contrast and by selecting salts with phosphate or sulfate anions, thereby adding strong scattering from P or S atoms. Low-dose electron diffraction of catalase crystals negatively stained with 200 mM magnesium sulfate shows Bragg spots extending out to 4.4 A. Future experimental use of sodium phosphate buffer and magnesium sulfate for negative staining is anticipated, particularly in designing new cocktail (multicomponent) negative stains able to support and protect protein structure to higher resolution levels than are currently achieved.
Massover
William H
WH
Department of Biological Sciences, Rutgers University-Newark, Newark, NJ 07102, USA. massover@andromeda.rutgers.edu
D017607:Aluminum Compounds / Q000737:chemistry; D000818:Animals; D002374:Catalase / Q000737:chemistry*; D002417:Cattle; D003460:Crystallization; D005293:Ferritins / Q000737:chemistry; D007773:Lactates / Q000737:chemistry; D008278:Magnesium Sulfate / Q000737:chemistry; D019565:Metals, Light / Q000737:chemistry*; D046529:Microscopy, Electron, Transmission / Q000379:methods; D016624:Negative Staining / Q000379:methods*; D012492:Salts / Q000737:chemistry*
D017607:Aluminum Compounds; D007773:Lactates; D019565:Metals, Light; D012492:Salts; D008278:Magnesium Sulfate; D005293:Ferritins; D002374:Catalase; C104223:aluminum lactate