494 Microsc. Microanal. 27 (Suppl 1), 2021
doi:10.1017/S1431927621002257 © Microscopy Society of America 2021
Application of the scanning ion-conductance microscopy (SICM) in study of
voriconazole impact on Candida parapsilosis surface structure.
Nikita Savin
1
, Vasilii Kolmogorov
2
, Roman Timoshenko
3
, Alexander Vaneev
4
, Aleksei Iakovlev
2
, Oleg
Suchalko
2
, Natalia Grammatikova
5
, Igor Levshin
5
, Natalia Klyachko
6
, Yurii Parkhomenko
7
, Sergei Salikhov
7
,
Alexander Majouga
8
, Alexander Erofeev
9
, Peter Gorelkin
9
and Yuri Korchev
1
1
National University of Science and Technology "MISIS", Moscow, Russia,
2
National University of Science
and Technology «MISiS», Moscow, Russia, Moskva, Russia,
3
National University of Science and Technology
"MISIS", Moscow, Moskva, Russia,
4
National University of Science and Technology «MISiS», Russia,
5
Gauze Research Institute for New Antibiotics, Moscow, Russian Federation, Russia,
6
Lomonosov Moscow
State University, Moscow, Russia, Russia,
7
NUST MISiS, Moscow, Russian Federation, Russia,
8
D.
Mendeleev University of Chemical Technology, Russia,
9
National University of Science and Technology
«MISiS», Moscow, Russia, Russia
The studying of antifungal drugs effect on the surface structure by traditional microbiological methods
is unobtainable. SICM method allows to obtain the topography of the surface structure and its mechanical
properties of biological samples in non-contact mode (Clarke et al., 2016). Achievable to obtain a data of cells
stiffness by applying pressure between of the nanopipette tip a sample surface. Azole drugs inhibit the synthesis
of ergosterol (a component of the yeast membrane) (Ghannoum & Rice, 1999), which lead to destruction and
softening of candida cell membrane. There are reports of the antifungal effect of caspofungin on elasticity of
cell wall (Quilès et al., 2017); impact of azole drugs on Candida surface structure (Madhavan et al., 2018;
Behbehani et al., 2019). However, the dynamic effect of azole drugs on the surface structure and mechanical
properties of Candida yeast in physiological conditions has not been studied. In this work, an impact of
fluconazole on Candida parapsilosis. To perform an experiment, yeast immobilized on a glutaraldehyde layer
was treated with voriconazole at 40 μg/ml for 6 hours. Cell morphology alteration of the Candida cell have
been observed by SICM (figure 1). In addition, displacement map indicates increase at 2.5 times (figure 2 (A,
B)). The average value of the displacement of the control cell was 17 nm, the value after cell treatment was 41
nm, which is indicated that the cell has softened after voriconazole treatment. Figure 2 (C) presents a graph of
the linear dependence of displacement value along the cell surface from the time of exposure with the drug.
The curve of the graph does not reach a plateau due to the short exposure time of cells with drug (less than 24
hours). An increase in displacement value indicates a decrease in stiffness at the cell surface and, consequently,
destruction of the Candida membrane. The data are consistent with theoretical concepts of the antifungal effect
of the azole drugs on Candida yeast. However, this characteristic is indirect in determining the mechanical
properties. Due to the relatively low pressure exerted on rigid yeast cells. Therefore, further refinement of the
method for determining cell elasticity is relevant.
This work was supported by the Russian Science Foundation grant No. 19-19-00626.
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