Phosphotungstate Staining of Vaccinia Virus

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Abstract
Vaccinia virus subjected to various repeated treatments and then negatively stained with potassium phosphotungstate was examined with the electron microscope. To obtain reproducible results it was necessary to standardize the preparation procedure. The standarized method now recommended is that the virus should be mixed with stain before being placed on the electron microscope grids and the grids then immediately examined or dried for 1 hr in a vacuum of at least 1 x 10-3 torr. The staining procedure may be altered to vary the degree of penetration of stain so as to reveal either internal or external detail.

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