Article contents
A Consumers Report on Detection of 10 Nm Gold on Cell Surfaces: Selection of Fesem Type and Choice of Conductive Metal Coating
Published online by Cambridge University Press: 02 July 2020
Extract
Important parameters for the detection of small colloidal immunogold particles (10 nm) on cell surfaces include the use of field emission SEM (FESEM) and a need for a conductive metal coating to reduce charging, so as to facilitate backscatter electron (BSE) imaging for unambiguous identification by atomic number contrast. Recently Heinzmann et al (1) have reported the use of below-lens FESEM for detection of 10 nm colloidal gold at working distances of 6-13 mm, but achieved success only on uncoated cells as coating with a 1 nm layer of platinum appeared to mask the gold signal. To improve specimen conductivity and signal collection, three metals (Cr, Pt, W) have been used to generate thin coatings either by planar magnetron sputtering for cryoSEM or ion-beam sputtering for routine chemically-fixed, critical point dried samples (2,3)
Three below-lens FESEM (Hitachi S-4500, JEOL 630land 63401, and LEO 982) were tested by using a sample of human neutrophils which had been chemically fixed, then immunostained for the surface antigen CD43 with 10 nm colloidal gold, and prepared by critical point drying.
- Type
- Low Voltage SEM Imaging and Analysis for the Biological and Materials Sciences
- Information
- Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 1231 - 1232
- Copyright
- Copyright © Microscopy Society of America 1997
References
- 1
- Cited by