Single Cell Level Microalgal green synthesis of silver nanoparticles: Confocal Microscopy and Digital Image Analysis

Year 2019, Volume: 2 Issue: 2, 56 - 60, 06.12.2019

Betül Yılmaz Öztürk Derviş Öztürk

Abstract

Nanoparticles are attracting
increasing attention due to their unusual and fascinating properties, which are
strongly influenced by their size, morphology and structure. Among the
developed nanoparticles, silver (Ag) nanoparticles are pertaining to have a
wide range of application in the fields of physical, chemical and biological
science. Physical
and chemical methods are used to synthesize such nanomaterials, among the
various known synthesis methods, biosynthesis of silver nanoparticles is
preferred as it is environmentally safe, low cost and less toxic. In
particular, the synthesis of nanoparticles in the cell can be achieved in a
standard size and shape. In the present work, the coccoid green algae Chodatodesmus mucronulatus was used as a reducing agent for the
synthesis of intracellular nanostructure silver particles (Ag-NPs). Algae are
with autofluorescence characteristics. These properties are known to be due to
chlorophyll pigments. In this context, a
confocal laser scanning microscopy (CLSM) based method to assess to show that
the amount of chlorophyll decreases at microalgae is reported. ). During this process, changes in
the amount of chlorophyll a, b and
carotenoid of the Chodatodesmus mucronulatus were examined at 24
hours using UV-Vis spectrophotometer for 3 days. As a result, the amount of
carotenoid, especially with the onset of the reaction, decreased markedly.
After 72 hours of reaction, the amount of carotenoid decreased from 6,54 μg ml^-1
to 0,00 μg / ml, chlorophyll a
decreased 24,46 µg ml^-1 to 0,06 µg ml^-1,chlorophyll b decreased from 11,33 µg ml^-1
to 4,15 µg /ml. This change (pigment amount in cells) was also observed with a
confocal microscope every 24 hours. Using this technique, the effect of in-use
concentrations of chlorophyll autofluorescence was defined. Determination of
mean fluorescence intensity (MFI) per cell by collecting auto-fluorescence from
single cells in x, y and z dimensions permitted evaluation at single-cell
level. According to the results, there is a decrease in the amount of pigment
in the cell. This suggests that the pigments may be capping agents and trigger
nanoparticle synthesis.

Keywords

Microalgae, Chlorophyll, Confocal microscopy, Nanoparticle, Silver

Thanks

This work was studied at Eskisehir Osmangazi University Central Research Laboratory Application and Research Center (ARUM).

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