Biosensing the Presence of Metal Nanoparticles by Spectrally- and Time-Resolved Endogenous Fluorescence in Water Moss Fontinalis antipyretica

Abstract

Pollution by heavy metals represents a significant environmental burden. We employed confocal microscopy with spectral detection and fluorescence lifetime imaging (FLIM) to evaluate the effect of nanoparticles (NPs) from various metals (Zinc, Nickel, Cobalt, Copper) on endogenous fluorescence of Fontinalis antipyretica moss. Short term (3–5 day) exposure to NPs, designed and fabricated by direct synthesis using femtosecond laser ablation in water, was studied. The green flavonoid and/or lignin endogenous fluorescence peaking between 500 and 560 nm was found to be increased by Zn and significantly reduced by Cu. The overall red chlorophyll fluorescence intensity with a maximum of 680 nm remained largely unchanged after exposure to Ni and Zn, but was decreased in the presence of Co and completely abolished by Cu. All NPs but Zn induced changes in the fluorescence lifetimes, demonstrating increased sensitivity of this parameter to environmental pollution. Gathered data indicate fast responsiveness of the endogenous fluorescence in the Fontinalis antipyretica moss to the presence of heavy metals that can thus potentially serve as a biosensing tool for monitoring environmental pollution in the moss natural environment.

Sensing the interaction of living organisms with microplastics by microscopy methods

“…Water bryophyte green moss species Fontinalis antipyretica was gathered commercially from aquashop (Aquasymbioza, Bratislava, Slovakia) and grown under ambient light. For the measurements, green moss fyloides were cut and placed on a microscope slide without further modification, as described previously [16].…”

“…Channel 1, recorded with BP 500-550 nm filter, served to monitor the fluorescently-labelled MPs and the endogenous fluorescence of the interstitial space. Channel 2, using BP 650-710 nm filter, was used to record the endogenous red chlorophyll fluorescence in the Fontinalis antipyretica moss, peaking at 680 nm [16]. Spectrally-resolved images of the moss in the presence or in the absence of MPs were taken across 7 spectral channels covering the spectral region from 472 nm to 643 nm with steps of 21.4 nm for the green, or covering the spectral region from 638 and 713 nm taken with 10.7 nm steps for the red chlorophyll fluorescence.…”

“…In this contribution, we evaluate the distribution of the custom-made fluorescently-labelled PS, PET, and PLA MPs and their effect on the endogenous fluorescence in the Fontinalis antipyretia moss. We have previously described the effect of heavy metals on the endogenous fluorescence in Fontinalis antipyretia moss [16], or Chlorella sp. algae [17].…”

Response of Coccomyxa cimbrica sp.nov. to Increasing Doses of Cu(II) as a Function of Time: Comparison between Exposure in a Microfluidic Device or with Standard Protocols

“…It has been extensively used to characterize structural changes and biochemical signaling in in vitro and in vivo experiments on many different types of cells [10][11][12] In plant cells and microorganisms, FLIM experiments have supplied information on the dynamic of the photosynthetic process [13,14]. Few FLIM experiments have also been devoted to the characterization of toxicants activity in microalgae [15,16] Almost all the studies assessing microalgae are based on in vitro experiments in which microalgae are cultivated in flasks or in microwells. In the last decade, microfluidic devices have shown to be an interesting alternative for the growth, manipulation, and characterization of cell cultures.…”

“…It has been extensively used to characterize structural changes and biochemical signaling in in vitro and in vivo experiments on many different types of cells [ 10 , 11 , 12 ] In plant cells and microorganisms, FLIM experiments have supplied information on the dynamic of the photosynthetic process [ 13 , 14 ]. Few FLIM experiments have also been devoted to the characterization of toxicants activity in microalgae [ 15 , 16 ]…”