Phosphorus imaging of the 7-S ribonucleoprotein particle

In this study an unprecedented demonstration of the detection sensitivity of electron spectroscopic imaging (ESI) is reported. This microanalytical technique is capable of forming elemental maps of a specimen with high sensitivity and resolution by forming images with electrons that have lost particular amounts of energy due to interactions with the atoms of the specimen. The 7-S ribonucleoprotein particle, composed of one molecule of 5-S RNA and one molecule of the 40K MW protein, TFIIIa, was used for this demonstration. As few as 120 phosphorus atoms of the 5-S RNA have been detected and their localization in the particle determined. The shape of the 7-S particle is ellipsoidal with a long axis of 15.0 nm and a shorter axis between 8 and 9 nm. Similarly, the 5-S RNA is also an elongated structure located asymmetrically on one side of the particle. The average signal-to-noise ratio over the particle in the net phosphorus images is 14 whereas the ratio measured for the nucleosome containing 2.4-fold more phosphorus is 30.