MAESTRO 10 NCN

Microfluidic methods for quantitative antibiotic susceptibility assays with single cell resolution (2018/30/A/ST4/00036)

Funding: National Science Centre, Poland

Grant period: 01.10.2019-30.09.2025

Project description:

Antibiotic resistant infections continue to be one of the grimmest threats to human health. The problem can only be effectively solved by better understanding of the response of bacteria to antibiotics. It will help to avoid emergence of resistance and to guide effective treatment.

That is why the objective of our project is to develop a new set of analytical tools that will allow to characterize the complexity of how bacteria respond to antibiotics used in clinical practice. To this end we use droplet microfluidics – a set of techniques that enable generation of thousands or millions of tiny, identical droplets. These droplets may be formed of a suspension of bacteria, resulting in encapsulation of single (individual) bacterial cells, or multiple cells in the droplets. By adding antibiotics at a range of concentrations, we are able to prepare sets (libraries) of droplets, each library containing droplets with a well-defined concentration of the antibiotic and number of bacterial cells.

Team members

  • Principal Investigator: prof. Piotr Garstecki
  • Postdocs: dr Ilona Foik, dr Marcin Równicki, dr Shahab Shahryari, dr Chirantan Das
  • Co-investigator: dr Paweł Jankowski
  • PhD students: Shakeel Ahmad, Kinga Głuchowska

Project dissemination

21.11. 2025

The doctoral defense of Shakeel Ahmad took place for the thesis “Deciphering the role of antibiotic exposure at the sub-minimum inhibitory concentration on bacterial heterogeneity at the single-cell level.” During the defense, the results obtained during the grant period were presented. The committee recommended to the Scientific Council of IChF that the dissertation be awarded a distinction.

07.04.2025

As part of the international Advancements in Microbiology conference held in Warsaw, Shakeel Ahmad delivered a 15-minute lecture titled “Sub-mic antibiotic pre-exposure increased the single-cell MIC and promoted hetero-resistance in bacteria” during the AMR & Pathogenesis symposium.
Ewelina Jałonicka gave a 5-minute flash talk, “Repositioning Cytostatics for Treating Opportunistic Bacterial Infections: The Microfluidic Approach to Using Cisplatin against Pseudomonas aeruginosa PA14.”
A post from the event tagged with NCN was published on LinkedIn and achieved a high reach.

16-18.05.2025

At the Warsaw University of Technology, the Symbiosis 2025 symposium took place, where Shakeel Ahmad delivered a 15-minute presentation titled “Antibiotic pre-exposure at sub-MIC level promotes heteroresistance between single cells inside microfluidic droplets.”

25-26.01.2022

Kinga Głuchowska, PhD student involved in the Maestro project, presented her research results at IChF PAN Microsympozjum. Her poster was entitled: „Quantitative, label-free and real-time monitoring of bacterial growth in nanoliter droplets”.

Publications

Shakeel Ahmad, Ilona P. Foik, Paweł Jankowski, Adam Samborski, Shreyas K. Vasantham, Piotr Garstecki. Ciprofloxacin pre-exposure influences individual cell MIC and heteroresistance of bacteria inside microfluidic droplets. Scientific Reports, 2025, 15, DOI: 10.1038/s41598-025-17157-0

Artur Ruszczak, Paweł Jankowski, Shreyas Vasantham, Ott Scheler, Piotr Garstecki. Physicochemical properties predict retention of antibiotics in water in oil droplets. Analytical Chemistry, 2023  https://doi.org/10.1021/acs.analchem.2c04644

Natalia Pacocha, Jakub Boguslawski, Karol Makuch, Marta Zapotoczna, Piotr Garstecki. You will know by its tail: a method for quantification of heterogeneity of bacterial populations using single-cell MIC profiling. Lab on a Chip, 2022 DOI: 10.1039/D2LC00234E

Witold Postek, Natalia Pacocha, Piotr Garstecki. Microfluidics for antibiotic susceptibility testing. Lab on a Chip 2022, Advance Article, published 07 Sep 2022.

Artur Ruszczak, Simona Bartkova, Marta Zapotoczna, Ott Scheler, Piotr Garstecki. Droplet-based methods for tackling antimicrobial resistance. Current Opinion in Biotechnology, Volume 76, August 2022.

Yu-Ting Kao, Silvia Calabrese, Nadine Borst, Michael Lehnert, Yu-Kai Lai, Franziska Schlenker, Peter Juelg, Roland Zengerle, Piotr Garstecki, Felix von Stetten. Microfluidic one-pot digital droplet FISH using LNA/DNA molecular beacons for bacteria detection and absolute quantification. Biosensors 2022, 12(4), 237.

Witold Postek, Piotr Garstecki. Droplet microfluidics for high-throughput analysis of antibiotic susceptibility in bacterial cells and populations. Accounts of Chemical Research, 2022, 55, 5, 605–615.

Dominika Ogończyk, Paweł Jankowski, Piotr Garstecki. A method for simultaneous polishing and hydrophobization of polycarbonate for microfluidic applications. Polymers, 2020, 12(11), 2490.