Abstract
EEG microstates, reflecting discrete topographical organization of the EEG signal power, may have clinical relevance; however, their functional significance and test-retest reliability remain unclear. To investigate the functional significance of the canonical EEG microstate classes and their pairwise transitions, and to establish their within-session test-retest reliability, we recorded 36-channel EEGs in 20 healthy volunteers during three eyes-closed conditions: mind-wandering, verbalization (silently repeating the word 'square' every 2 s), and visualization (visualizing a square every 2 s). Each condition lasted 3 min and the sequence of three conditions was repeated four times (two runs of two sequence repetitions). The participants' alertness and their sense of effort during the experiment were rated using visual-analogue scales. The EEG data were 2-20 Hz bandpass-filtered and analysed into the four canonical microstate classes: A, B, C, and D. EEG microstate classes C and D were persistently more dominant than classes A and B in all conditions. Of the first-order microstate parameters, average microstate duration was most reliable. The duration of class D microstate was longer during mind-wandering (106.8 ms) than verbalization (102.2 ms) or visualization (99.8 ms), with a concomitantly higher coverage (36.4% vs. 34.7% and 35.2%), but otherwise there was no clear association of the four microstate classes to particular mental states. The test-retest reliability was higher at the beginning of each run, together with higher average alpha power and subjective ratings of alertness. Only the transitions between classes C and D (from C to D in particular) were significantly higher than what would be expected from the respective microstates' occurrences. The transition probabilities, however, did not distinguish between conditions, and their test-retest reliability was overall lower than that of the first-order parameters such as duration and coverage. Further studies are needed to establish the functional significance of the canonical EEG microstate classes. This might be more fruitfully achieved by looking at their complex syntax beyond pairwise transitions. To ensure greater test-retest reliability of microstate parameters, study designs should allow for shorter experimental runs with regular breaks, particularly when using EEG microstates as clinical biomarkers.