A technique is described, which is new in certain details, for fracturing frozen cells and taking replicas from the fracture surface. This technique has been developed as a cytological preparative technique complementary to those currently used in electron microscopy. The present paper describes results for human red cells, chosen for preliminary work to facilitate recognition of artifacts. The cells are suspended in 20 per cent glycerol, 0.9 per cent NaCl solution, allowed time to equilibrate, and rapidly frozen in a small chamber plunged into propane at -196°C. The chamber is kept at -196°C., and fractured in vacuo, in such a way that the fracture plane runs through the frozen suspension, and through the cells in its path. After fracture, the chamber is brought to -105°C., still in vacuo, to etch the fracture surface, and a platinum-carbon replica is deposited on the surface from a carbon arc. The replica is subsequently cleaned in concentrated alkaline solution and examined in the electron microscope. The outlines of the fractured cells can be recognised. There are indications that in areas of the replica which correspond to the interior of a cell, individual haemoglobin molecules can be seen, and in favourable cases the arrangement of some of the four molecular subunits.
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1 April 1961
Content prior to 1962 was published under the journal name
The Journal of Biophysical and Biochemical Cytology
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April 01 1961
ELECTRON MICROSCOPE REPLICAS FROM THE SURFACE OF A FRACTURE THROUGH FROZEN CELLS
G. H. Haggis
G. H. Haggis
From the Physiology Department, University of Edinburgh, Scotland
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G. H. Haggis
From the Physiology Department, University of Edinburgh, Scotland
Received:
September 07 1960
Copyright, 1961, by The Rockefeller Institute Press
1961
J Biophys and Biochem Cytol (1961) 9 (4): 841–852.
Article history
Received:
September 07 1960
Citation
G. H. Haggis; ELECTRON MICROSCOPE REPLICAS FROM THE SURFACE OF A FRACTURE THROUGH FROZEN CELLS . J Biophys and Biochem Cytol 1 April 1961; 9 (4): 841–852. doi: https://doi.org/10.1083/jcb.9.4.841
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