Semi-automated colony-forming unit counting for biosafety level 3 laboratories.

High Throughput Screening Microbiology

Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
06 Aug 2023
Historique:
received: 20 03 2023
revised: 10 05 2023
accepted: 15 06 2023
medline: 7 8 2023
pubmed: 7 8 2023
entrez: 7 8 2023
Statut: aheadofprint

Résumé

Biosafety level 3 decontamination precautions motivate measuring microbial colonies using consumable photography instead of expensive automated plate counters or smartphones, and assaying drug treatments-with multiple concentrations per treatment, replicates, and controls-produces hundreds of images. Here, we present a protocol for semi-automated image analysis by hand-tuning three parameters. The parameters control for non-uniform colony growth and artifacts such as lid condensation, reflections, and plating streaks. We describe steps to prepare images, tune parameters, and plot dose-response relationships. For complete details on the use and execution of this protocol, please refer to Larkins-Ford et al.

Identifiants

pubmed: 37549035
pii: S2666-1667(23)00409-4
doi: 10.1016/j.xpro.2023.102442
pmc: PMC10424145
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

102442

Subventions

Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : NIAID NIH HHS
ID : R01 AI150684
Pays : United States

Informations de copyright

Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests The authors declare no competing interests.

Références

Nat Methods. 2012 Jun 28;9(7):676-82
pubmed: 22743772
Bioinformatics. 2016 Oct 15;32(20):3210-3212
pubmed: 27354701
Cell Syst. 2021 Nov 17;12(11):1046-1063.e7
pubmed: 34469743
BMC Bioinformatics. 2021 Sep 10;22(1):433
pubmed: 34507520

Auteurs

Soorya Janakiraman (S)

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109, USA.

Sophie Engels (S)

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109, USA.

Pariksheet Nanda (P)

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109, USA.

Maral Budak (M)

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109, USA.

Talia Greenstein (T)

Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA 02111, USA; Department of Biomedical Engineering, Tufts University School of Engineering, Medford, MA 02155, USA.

Mariana Pereira Moraes (MP)

Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA 02111, USA; Department of Biomedical Engineering, Tufts University School of Engineering, Medford, MA 02155, USA.

Bree B Aldridge (BB)

Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA 02111, USA; Department of Biomedical Engineering, Tufts University School of Engineering, Medford, MA 02155, USA.

Denise E Kirschner (DE)

Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109, USA. Electronic address: kirschne@umich.edu.

Classifications MeSH