Transient freezing behavior in photophobic responses of Euglena gracilis investigated in a microfluidic device.

Abstract

We found that the transient freezing behavior in photophobic responses of Euglena gracilis is a good indicator of the metabolic status of the cells. The transient blue light photophobic responses of E. gracilis cells were investigated on-chip using a new measurement, 'trace momentum' (TM), to evaluate their swimming activity quantitatively in real time. When blue light of intensity >30 mW cm(-2) was repeatedly switched on and off, a large negative spike in the TM was observed at the onset of the 'blue-light-off' phase. Single-cell trace analysis at a blue light intensity of 40 mW cm(-2) showed that 48% (on average, n = 15) of tumbling Euglena cells ceased activity ('freezing') for 2-30 s at the onset of blue-light-off before commencing forward motion in a straight line (termed 'straightforward swimming'), while 45% smoothly commenced straightforward swimming without delay. The proportion of freezing Euglena cells depended on the blue light intensity (only 20% at 20 mW cm(-2)). When the cells were stimulated by four blue light pulses at the higher intensity, without pre-exposure, the transient freezing behavior was more prominent but, on repeating the stimuli after an 80 min interval in red light, the same cells did not freeze. This shows that the metabolism of the cells had changed to anti-freezing during the interval. The relationship between the interval time with/without light irradiation and the blue light adaptation was elucidated experimentally. The origin of the freezing behavior is considered to be a shortage of a metabolic substance that promotes smooth switching of flagellum movement from in situ rotation mode to a straightforward swimming mode.