Cell-surface receptors and proteins on platelet membranes imaged by scanning force microscopy using immunogold contrast enhancement

S. J. Eppell, S. R. Simmons, R. M. Albrecht, R. E. Marchant

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

High resolution scanning force microscope (SFM) images of fibrinogen-exposed platelet membranes are presented. Using ultrasharp carbon tips, we are able to obtain submolecular scale resolution of membrane surface features. Corroboration of SFM results is achieved using low voltage, high resolution scanning electron microscopy (LVHRSEM) to image the same protein molecule that is seen in the SFM. We obtain accurate height dimensions by SFM complemented by accurate lateral dimensions obtained by LVHRSEM. The use of 14- and 5-nm gold labels to identify specific membrane-bound biomolecules and to provide contrast enhancement with the SFM is explored as a useful adjunct to observation of unlabeled material. It is shown that the labels are useful for locating specific protein molecules on platelet membrane surfaces and for assessing the distribution of these molecules using the SFM. Fourteen nm labels are shown to be visible over the membrane corrugation, whereas 5-nm labels appear difficult to resolve using the present SFM instrumental configuration. When using the 5-nm labels, collateral use of LVHRSEM allows one to examine SFM images at submolecular resolution and associate function with the structures imaged after the SFM experiment is completed.

Original languageEnglish
Pages (from-to)671-680
Number of pages10
JournalBiophysical Journal
Volume68
Issue number2
DOIs
StatePublished - 1995
Externally publishedYes

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