Eight modifications of hemagglutination technique were applied to the sensitization of sheep erythrocytes with Au antigen; these included tannic acid (TA), bis-diazotized benzidine (BDB), tolylene-2, 4-diisocyanate (TDIC), difluorodinitrobenzene, tetraazotized O-dianisidine (TOD), cyanulic chloride5), glutaraldehyde and chromic chloride (CrCl₃) method.
The sensitized cells could be obtained only using TA, BDB, TDIC or TOD in the glutaraldehyde-treated cells and CrCl₃ in the native cells, if the concentration of Au antigen in the reaction solution was limited to about 25γ/ml. Among these reagents the best result was obtained with BDB.
The coupling of Au antigen to erythrocytes with these reagents seems not significantly to inactivate the antigenic groups of Au antigen.
The cells coated with the aid of BDB, TDIC or TA could be used again without any appreciable loss of hemagglutination titer for 4 months when preserved at 4°C.

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It is well known that physical and chemical changes occur during storage of preserved blood. Those changes, , however, depend on storage conditions. For example, there are differences depending on whether bloom is stored in glass bottles or in plastic bags. One of the main reasons for differences under these conditions seems to be oxygenation effect in the plastic bags. Preserved blood stored in plastic bags takes up oxygen through the bags. We have studied this oxygenation effect on preserved blood before and after airation in glass bottles and in plastic bags.
The changes in the following properties were measured:
(1) Osmotic fragility of red blood cells.
(2) Hemoglobin content in plasma.
(3) Viscosity of whole blood.
(4) pH in plasma.
(5) ATP and 2, 3-DPG contents in red blood cells.
There are no significant changes in the properties from (1) to (4) after 42 days in blood stored in plastic bags, while big changes have been seen in blood stored in glass bottles after 14 days.
ATP and 2, 3-DPG have been consumed in both cases, but consumption was much higher in blood stored in glass bottles. Blood stored in glass bottles after airation has shown a pattern close to that of stored in plastic bags.
In conclusion, plastic bags cause less changes in the properties mentioned above. It is clear that oxygen strongly affects preserved blood as expected.

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