Hostname: page-component-84b7d79bbc-rnpqb Total loading time: 0 Render date: 2024-07-30T06:12:31.534Z Has data issue: false hasContentIssue false

The Utilization of Cryopreservation in Small Gold Surface Labeling

Published online by Cambridge University Press:  02 July 2020

C.A. Ackerley
Affiliation:
Division of Pathology, Hospital for Sick Children, Toronto, Ont., Canada, M5G1X8
A. Tilups
Affiliation:
Division of Pathology, Hospital for Sick Children, Toronto, Ont., Canada, M5G1X8
T. Kanazawa
Affiliation:
JEOL USA Inc., Peabody, MA01960
L.E. Becker
Affiliation:
Division of Pathology, Hospital for Sick Children, Toronto, Ont., Canada, M5G1X8
Get access

Extract

Cryopreservation has many advantages over routine chemical fixation. Two of these are the almost spontaneous arrest of biological activity and the retention of material that normally would be lost with chemical fixation and dehydration. In the current study 2 systems were evaluated. Hyaluronic acid (HA) was covalently bound to 1.4nm nanogold® and applied to astrocytoma cell cultures and the uptake monitored over time using both cryopreparative methods and conventional chemical fixation. The second system evaluated was the detection of liposomes containing CFTR covalently bound to 1.4nm gold in the airways of mice from a phenotype of an inbred congenic strain of CFTR-knockout mice that spontaneously develop lung disease.

In the HA uptake experiment, astrocytoma cell cultures were treated with HA gold for either 5, 10 or 20 minutes. At the appropriate time point, cells were either plunge frozen in liquid ethane or fixed with 2.5% glutaraldehyde in phosphate buffer.

Type
Low Temperature Methods for Immunolabeling of Cells and Tissues
Copyright
Copyright © Microscopy Society of America

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

1.Kent, G. et al., J. Clin. Invest. 1997 100:30603069CrossRefGoogle Scholar
2.Ramjeesingh, M. et al., Human Gene Therapy 1998 9:521528CrossRefGoogle Scholar
3.Ackerley, C.A. et al., Proc., Microscopy and Microanalysis 1999 484485CrossRefGoogle Scholar