Analysis of high quality monatomic chromium films used in biological high resolution scanning electron microscopy.

During the recent employment of field emission (FE) in-lens scanning electron microscopes (SEMs), refractory metal deposition technology has co-evolved to provide enhanced contrast of 1-10 nm hydrocarbon based biological structures imaged at high magnifications (> 200,000 times). Pioneer development employing the Penning sputter system in a high vacuum chamber proved that imaging of chromium (Cr) coated biological specimens contained enriched secondary electron (SE)-(I) contrasts. Single nanometer size fibrillar and particulate ectodomains within the context of complex biological membranes were accurately imaged without significant enlargement using the high resolution SE-I mode (HRSEM). This paper reports the transmission electron microscopy (TEM) testing of ultrathin (0.5-2.0 nm) Cr films deposited by planar magnetron sputter coating (PMSC). Essential parameters necessary to reproduce quality sputtered films of refractory metals used in HRSEM studies were described for the vacuum system and target operation conditions (current, voltage, and target distance). HRSEM imaging of biological specimens is presented to assess contrast attained from ultrathin fine grain Cr films deposited by PMSC. High magnification images were recorded to illustrate high quality contrasts attainable by HRSEM at low (1-5 kV) and high (10-30 kV) voltages. Dispersed molecules on formvar coated grids were sputter coated with a 1 nm thick Cr film before employing scanning transmission (STEM)/SEM modes of the FESEM to establish non-decorative image accuracy in the transmitted electron mode.