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The Utilization of Cryopreservation in Small Gold Surface Labeling

Published online by Cambridge University Press:  02 July 2020

C.A. Ackerley ,
A. Tilups ,
T. Kanazawa  and
L.E. Becker
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
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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
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 304 - 305
Copyright
Copyright © Microscopy Society of America

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References

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