Effect of Pumping Pulse Duration on Echo Signal Amplitude in Four-Pulse PELDOR

The impact of pumping pulse duration on four-pulse pulsed electron–electron double resonance (PELDOR) data was experimentally studied. For biradicals with known distances between two spin labels, it is shown that refocused echo amplitude decreases with increasing the pumping pulse duration and decreasing the distance between spin labels. The effect becomes substantial when the pumping pulse duration is comparable or exceeds the inverse value of the dipole–dipole interaction between spin labels. This effect is essential for determination of distance distribution between labels in double-labeled molecules and for determination of the number of labels in clusters of spin-labeled molecules. PELDOR signal distortion was observed when the pumping pulse position in the time scale coincided with those of the detecting pulses. An approach of signal correction to eliminate this distortion is proposed.