CEEPC/IPM/CMSC - Abstrakt prezentace

(5. konference České společnosti pro hmotnostní spektrometrii - WeP-030)
High resolution imaging of Aspergillus fumigatus infection

Tomáš Pluháček 1,2 *, Miloš Petřík 3, Dominika Luptáková 1, Oldřich Benada 1, Andrea Palyzová 1, Karel Lemr 1,2, Vladimír Havlíček 1,2

  1. Institute of Microbiology ASCR, Prague
  2. RCPTM, Department of Analytical chemistry, Faculty of Science, Palacký University, Olomouc
  3. IMTM, Faculty of Medicine and Dentistry, Palacký University, Olomouc

Abstrakt

Aspergillus fumigatus, an airborne saprophytic fungus, is being a leading cause of different opportunistic pulmonary infections, comprising aspergilloma, invasive pulmonary aspergillosis (IPA), allergic bronchopulmonary aspergillosis, chronic pulmonary aspergillosis, chronic necrotizing aspergillosis and allergic fungal rhinositis. In immune-compromised patients the IPA is rapidly progressive disease with the short survival time, which accounts for more than 200,000 infections per year, with an associated mortality rate of 30 - 90 % [1,2].

In this sense, presented work has been focused on monitoring of disseminated aspergillosis in rat lungs via the gold and silver LA-ICP-MS imaging employing modified Grocott’s methenamine silver staining and eosin counterstaining. Analysis of the gold and silver was carried out by the excimer laser ablation system Analyte G2 (Photon Machines, USA) coupled to a quadrupole ICP-MS spectrometer (ORS-ICP-MS Agilent 7700x, Agilent, Japan). The silver and gold distribution maps collected with the 5 μm laser focus provided reliable images of fungal hyphae deposition in tissues, so both can be used as excellent Aspergillus infection tracers. The limit of detection was determined for both 107Ag and 197Au as 0.03 μg.g−1. The selective incorporation of Ag and Au into microbial hyphae and their low background noise were confirmed by energy dispersive X-ray scattering utilizing the submicron lateral resolving power [3].

In conclusion, the gold and silver LA-ICP-MS imaging with the lateral resolution getting close to the fungal hyphae diameter has a substantial potential to become a more sensitive alternative to current standard visualization tools used in clinical mycology.

* Korespondující autor: pluhacektomas20@seznam.cz

Literatura

  1. Pluhacek T. et al: Mass Spectrom. Rev. 35, 35-47 (2016).
  2. Thornton C. R.: Expert. Rev. Immunol. 10, 771-780 (2014).
  3. Pluhacek T. et al: submitted

Poděkování:

The authors gratefully acknowledge the support from the Ministry of Education, Youth and Sport of the Czech Republic (LO1305, LO1509) and Czech Science Foundation (P206/12/1150).


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