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Autofluorescence Imaging to Evaluate Red Algae Physiology

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Time-resolved endogenous chlorophyll fluorescence sensitivity to pH: study on Chlorella sp. algae.

A Marcek Chorvatova¹, M Uherek, A Mateasik

¹Department of Biophotonics, International Laser Centre, Ilkovicova 3, 84104 Bratislava, Slovakia. Department of Biophysics, Faculty of Natural Sciences, University of Ss. Cyril and Methodius, nam. J Herdu 2, 91701 Trnava, Slovakia.

Methods and Applications in Fluorescence

|February 20, 2020

Summary

Algae

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Area of Science:

Algal biology
Spectroscopy
Microscopy

Background:

Endogenous fluorescence in algae is influenced by environmental factors.
Chlorophylls are key fluorescent compounds in algae.
Understanding pH effects on algal fluorescence is crucial for ecological monitoring.

Purpose of the Study:

To investigate the pH-dependence of endogenous fluorescence in green algae (Chlorella sp.).
To explore the potential of algal fluorescence as a pH biosensing tool.

Main Methods:

Utilized spectroscopy and advanced time-resolved fluorescence imaging microscopy (FLIM).
Employed picosecond pulsed laser excitation at 450 nm and 475 nm.
Analyzed absorption and emission spectra, fluorescence intensity, and fluorescence lifetimes (tau1, tau2).

Main Results:

Acidification decreased red fluorescence (680 nm) intensity and shortened tau1 lifetime.
Severe acidification increased green fluorescence (520-540 nm) and shifted red fluorescence (690-700 nm).
Observed prolongation of tau2 fluorescence lifetime under severe acidification.

Conclusions:

Algal fluorescence, particularly from chlorophylls, shows significant pH-dependence.
Changes in fluorescence intensity and lifetime can indicate environmental pH shifts.
Algal endogenous fluorescence is a promising candidate for developing novel pH biosensors.