Label-free metabolic imaging and energy costs in Chlamydomonas.

Abstract

We developed a label-free optical microscopy method to study movements of different frequencies and amplitudes within a cell. We use optical transmission tomography (OTT) that operates in transmission, and we record the changes of signal values of all the pixels of movies taken for a few seconds (dynamic signal). This signal is a metabolic signal in algae as it decreased in the presence of photosystem II inhibitors or when samples were illuminated at wavelengths where the photoreceptors are poorly operative. We used as model organism Chlamydomonas for which mutants are available. We used a mutant deleted of the chloroplastic gene encoding the large subunit of the Rubisco, ΔrbcL. This mutant is unable to fix atmospheric CO₂ and is devoid of pyrenoid. We compared the dynamic signal between wild-type strain and ΔrbcL mutant of Chlamydomonas grown in dark condition and found it to be 5 to 10 times higher. This mutant overproduced starch, and we tempted to associate the metabolic signal to the cost in ATP_eq consumption for building starch. The method is easy to implement and could be very valuable for studies of phytoplankton in situ or virus-infected cells.