A low-cost DIY device for high resolution, continuous measurement of microbial growth dynamics

High-resolution data on microbial growth dynamics allow characterisation of microbial physiology, as well as optimisation of genetic alterations thereof. Such data are routinely collected using bench-top spectrophotometers or so-called plate readers. These equipments present several drawbacks: (i) measurements from different devices cannot be compared directly, (ii) proprietary nature of devices makes it difficult for standardisation methods to be developed across devices, and (iii) high costs limit access to devices, which can become a bottleneck for researchers, especially for those working with anaerobic organisms or at higher containment level laboratories. These limitations could be lifted, and data reproducibility improved, if the scientific community could adopt standardised, low-cost and open-source devices that can be built in-house. Here, we present such a device, MicrobeMeter, which is a do-it-yourself (DIY), simple, yet robust photometer with continuous data-logging capability. It is built using 3D-printing and open-source Arduino platform, combined with purpose-built electronic circuits. We show that MicrobeMeter displays linear relation between culture density and turbidity measurement for microbes from different phylogenetic domains. In addition, culture density estimated from MicrobeMeter measurements produced less variance compared against three commercial bench-top spectrophotometers, indicating that its measurements are less affected by the differences in cell types. We show the utility of MicrobeMeter, as a programmable wireless continuous measurement device, by collecting long-term growth dynamics up to 458 hours from aerobic and anaerobic cultures. We provide a full open-source description of MicrobeMeter and its implementation for faster adaptation and future development by the scientific community. The blueprints of the device, as well as ready-to-assemble kit versions are also made available through www.humanetechnologies.co.uk.

[1]  A. L. Koch,et al.  Turbidity measurements of bacterial cultures in some available commercial instruments. , 1970, Analytical biochemistry.

[2]  Dean Calloway,et al.  Beer-Lambert Law , 1997 .

[3]  J. Monod The Growth of Bacterial Cultures , 1949 .

[4]  Joshua M. Pearce,et al.  Building Research Equipment with Free, Open-Source Hardware , 2012, Science.

[5]  Walt Kester,et al.  SECTION 7-4 – Op Amp Protection , 2005 .

[6]  R. Cross,et al.  Curvature-induced expulsion of actomyosin bundles during cytokinetic ring contraction , 2016, eLife.

[7]  M. Meyer Domesticating and democratizing science: A geography of do-it-yourself biology , 2013 .

[8]  R. Maier Bacterial Growth , 2008 .

[9]  J. Myers,et al.  Improving accuracy of cell and chromophore concentration measurements using optical density , 2013, BMC biophysics.

[10]  S. Leibler,et al.  Contingency and Statistical Laws in Replicate Microbial Closed Ecosystems , 2012, Cell.

[11]  G. Bertani,et al.  STUDIES ON LYSOGENESIS I , 1951, Journal of bacteriology.

[12]  Orkun S. Soyer,et al.  A stable genetic polymorphism underpinning microbial syntrophy , 2016, The ISME Journal.

[13]  J. Lawrence,et al.  Correction for the inherent error in optical density readings , 1977, Applied and environmental microbiology.

[14]  Joseph McFARLAND,et al.  THE NEPHELOMETER:AN INSTRUMENT FOR ESTIMATING THE NUMBER OF BACTERIA IN SUSPENSIONS USED FOR CALCULATING THE OPSONIC INDEX AND FOR VACCINES. , 1907 .

[15]  Sara Marques,et al.  Simple and Versatile Turbidimetric Monitoring of Bacterial Growth in Liquid Cultures Using a Customized 3D Printed Culture Tube Holder and a Miniaturized Spectrophotometer: Application to Facultative and Strictly Anaerobic Bacteria , 2016, Front. Microbiol..

[16]  A. L. Koch,et al.  Th size and shape of bacteria by light scattering measurements. , 1968, Biochimica et biophysica acta.

[17]  Peter S. Swain,et al.  General calibration of microbial growth in microplate readers , 2016, Scientific Reports.

[18]  Effects of cellular fine structure on scattered light pattern , 2006, IEEE Transactions on NanoBioscience.