Utilizing atomic number contrast for FESEM imaging of colloidal nanotopography underlying biological cells

Authors: Wood, M.A.; Meredith, D.O.; Owen, G Rh; Richards, R.G.; Riehle, M.O.

Source: Nanotechnology, Volume 16, Number 9, September 2005 , pp. 1433-1439(7)

Publisher: Institute of Physics Publishing

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Abstract:

Developments within the electronics industry have led to the fabrication of surfaces with in-plane nanofeatures for use in biological investigations. An inexpensive, high throughput and accessible method has evolved, where a natural lithography technique utilizes off-the-shelf colloidal particles within the nanometric range to produce a variety of patterned substrates. Imaging of cell interactions with respect to underlying nanotopography (sub-100 nm) requires high resolution microscopy techniques. When utilizing field emission scanning electron microscopy (FESEM), operation is restricted to low voltage conditions in an attempt to image fine surface detail while limiting radiation damage, charging and carbon deposition with respect to biological material. The maximum number of backscattered electrons (BSEs) produced from biological specimens is achieved using low accelerating voltages while the number of BSEs emitted from high atomic number materials increases over the accelerating voltage range of 2–16 keV. By increasing the emission current, the number of electrons present in the primary beam is elevated, increasing BSE production at the low voltages utilized. Through the careful selection of high atomic number materials composing the colloids or similar topographical features, secondary electron (SE) and BSE detection can be utilized as complementary imaging techniques. This allows for the surface morphology of cell membranes to be imaged by SE detection in relation to the underlying high contrast nanotopography where BSEs are collected. Furthermore, cytoskeletal features of varying density can also be identified in relation to cells and colloidal nanofeatures when detecting BSEs.

Document Type: Research article

DOI: 10.1088/0957-4484/16/9/002

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