Mapping of nutrient-induced biochemical changes in living algal cells using synchrotron infrared microspectroscopy.
High quality Fourier transform infrared (FTIR) spectra were acquired from living Micrasterias hardyi cells maintained in an IR transparent flow-through cell using a FTIR microscope coupled to a synchrotron light source. Spectral maps of living, nutrient-replete cells showed band intensities consistent with the known location of the nucleus and the chloroplasts. These were very similar to maps acquired from fixed, air-dried cells. Bands due to lipids were lowest in absorbance in the region of the nucleus and highest in the chloroplast region and this trend was reversed for the absorbance of bands attributed to protein. Spectra acquired in 10 microm steps across living phosphorus-starved (P-starved) cells, repeated approximately every 30 min, were consistent over time, and bands correlated well with the known position of the nucleus and the observed chloroplasts, corroborating the observations with replete cells. Experiments in which missing nutrients were re-supplied to starved cells showed that cells could be maintained in a functional state in the flow-through cell for up to one day. Nitrogen-starved cells re-supplied with N showed an increase in lipid in all positions measured across the cell over a 23 h period of re-supply, with the largest increases occurring in positions where the chloroplasts were observed. Re-supply of phosphorus to P-starved cells produced no changes in bands attributable to lipid or protein. Due to their thin cell body ( approximately 12 microm) and large diameter ( approximately 300 microm) Micrasterias sp. make an ideal spectroscopic model to study nutrient kinetics in algal cells.
Heraud
Philip
P
Centre for Biospectroscopy and School of Chemistry, Monash University, 3800 Vic., Australia. phil.heraud@sci.monash.edu.au
Wood
Bayden R
BR
Tobin
Mark J
MJ
Beardall
John
J
McNaughton
Don
D
D056890:Eukaryota / Q000254:growth & development*; D050356:Lipid Metabolism; D011506:Proteins / Q000378:metabolism; D013055:Spectrophotometry, Infrared; D017550:Spectroscopy, Fourier Transform Infrared; D017356:Synchrotrons
D011506:Proteins