PMID- 35558922 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20220519 IS - 2046-2069 (Electronic) IS - 2046-2069 (Linking) VI - 8 IP - 64 DP - 2018 Oct 26 TI - Identification of pathogenic bacteria in complex samples using a smartphone based fluorescence microscope. PG - 36493-36502 LID - 10.1039/c8ra06473c [doi] AB - Diagnostics based on fluorescence imaging of biomolecules is typically performed in well-equipped laboratories and is in general not suitable for remote and resource limited settings. Here we demonstrate the development of a compact, lightweight and cost-effective smartphone-based fluorescence microscope, capable of detecting signals from fluorescently labeled bacteria. By optimizing a peptide nucleic acid (PNA) based fluorescence in situ hybridization (FISH) assay, we demonstrate the use of the smartphone-based microscope for rapid identification of pathogenic bacteria. We evaluated the use of both a general nucleic acid stain as well as species-specific PNA probes and demonstrated that the mobile platform can detect bacteria with a sensitivity comparable to that of a conventional fluorescence microscope. The PNA-based FISH assay, in combination with the smartphone-based fluorescence microscope, allowed us to qualitatively analyze pathogenic bacteria in contaminated powdered infant formula (PIF) at initial concentrations prior to cultivation as low as 10 CFU per 30 g of PIF. Importantly, the detection can be done directly on the smartphone screen, without the need for additional image analysis. The assay should be straightforward to adapt for bacterial identification also in clinical samples. The cost-effectiveness, field-portability and simplicity of this platform will create various opportunities for its use in resource limited settings and point-of-care offices, opening up a myriad of additional applications based on other fluorescence-based diagnostic assays. CI - This journal is (c) The Royal Society of Chemistry. FAU - Muller, Vilhelm AU - Muller V AUID- ORCID: 0000-0002-4554-2124 AD - Department of Biology and Biological Engineering, Chalmers University of Technology Gothenburg Sweden fredrikw@chalmers.se. FAU - Sousa, Jose M AU - Sousa JM AD - Biomode 2, S.A., Edificio GNRation Praca Conde Agrolongo no. 123 4700-312 Braga Portugal. FAU - Ceylan Koydemir, Hatice AU - Ceylan Koydemir H AD - Electrical and Computer Engineering Department, University of California Los Angeles 90095 CA USA ozcan@ucla.edu. AD - Bioengineering Department, University of California Los Angeles 90095 CA USA. AD - California NanoSystems Institute (CNSI), University of California Los Angeles 90095 CA USA. FAU - Veli, Muhammed AU - Veli M AD - Electrical and Computer Engineering Department, University of California Los Angeles 90095 CA USA ozcan@ucla.edu. AD - Bioengineering Department, University of California Los Angeles 90095 CA USA. AD - California NanoSystems Institute (CNSI), University of California Los Angeles 90095 CA USA. FAU - Tseng, Derek AU - Tseng D AD - Electrical and Computer Engineering Department, University of California Los Angeles 90095 CA USA ozcan@ucla.edu. AD - Bioengineering Department, University of California Los Angeles 90095 CA USA. AD - California NanoSystems Institute (CNSI), University of California Los Angeles 90095 CA USA. FAU - Cerqueira, Laura AU - Cerqueira L AD - Biomode 2, S.A., Edificio GNRation Praca Conde Agrolongo no. 123 4700-312 Braga Portugal. AD - LEPABE, Department of Chemical Engineering, Faculty of Engineering of the University of Porto Rua Dr Roberto Frias 4200-465 Porto Portugal nazevedo@fe.up.pt. FAU - Ozcan, Aydogan AU - Ozcan A AUID- ORCID: 0000-0002-0717-683X AD - Electrical and Computer Engineering Department, University of California Los Angeles 90095 CA USA ozcan@ucla.edu. AD - Bioengineering Department, University of California Los Angeles 90095 CA USA. AD - California NanoSystems Institute (CNSI), University of California Los Angeles 90095 CA USA. FAU - Azevedo, Nuno F AU - Azevedo NF AD - LEPABE, Department of Chemical Engineering, Faculty of Engineering of the University of Porto Rua Dr Roberto Frias 4200-465 Porto Portugal nazevedo@fe.up.pt. FAU - Westerlund, Fredrik AU - Westerlund F AUID- ORCID: 0000-0002-4767-4868 AD - Department of Biology and Biological Engineering, Chalmers University of Technology Gothenburg Sweden fredrikw@chalmers.se. LA - eng PT - Journal Article DEP - 20181029 PL - England TA - RSC Adv JT - RSC advances JID - 101581657 PMC - PMC9088845 COIS- N. F. A. and L. C. are co-founders and shareholders of Biomode, a company that specializes in the development of PNA-FISH methods for the detection of microorganisms. A. O. has pending patent applications on mobile phone-based microscopy technologies. EDAT- 2018/10/29 00:00 MHDA- 2018/10/29 00:01 PMCR- 2018/10/29 CRDT- 2022/05/13 18:21 PHST- 2018/08/01 00:00 [received] PHST- 2018/10/21 00:00 [accepted] PHST- 2022/05/13 18:21 [entrez] PHST- 2018/10/29 00:00 [pubmed] PHST- 2018/10/29 00:01 [medline] PHST- 2018/10/29 00:00 [pmc-release] AID - c8ra06473c [pii] AID - 10.1039/c8ra06473c [doi] PST - epublish SO - RSC Adv. 2018 Oct 29;8(64):36493-36502. doi: 10.1039/c8ra06473c. eCollection 2018 Oct 26.