Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Silicon photon-counting detector for full-field CT using an ASIC with adjustable shaping time

Purpose: Photon-counting silicon strip detectors are attracting interest for use in next-generation CT scanners. For CT detectors in a clinical environment, it is desirable to have a low power consumption. However, decreasing the power consumption leads to higher noise. This is particularly detrimental for silicon detectors, which require a low noise floor to obtain a good dose efficiency. The increase in noise can be mitigated using a longer shaping time in the readout electronics. This also results in longer pulses, which requires an increased deadtime, thereby degrading the count-rate performance. However, as the photon flux varies greatly during a typical CT scan, not all projection lines require a high count-rate capability. We propose adjusting the shaping time to counteract the increased noise that results from decreasing the power consumption. Approach: To show the potential of increasing the shaping time to decrease the noise level, synchrotron measurements were performed using a detector prototype with two shaping time settings. From the measurements, a simulation model was developed and used to predict the performance of a future channel design. Results: Based on the synchrotron measurements, we show that increasing the shaping time from 28.1 to 39.4 ns decreases the noise and increases the signal-to-noise ratio with 6.5% at low count rates. With the developed simulation model, we predict that a 50% decrease in power can be attained in a proposed future detector design by increasing the shaping time with a factor of 1.875. Conclusion: Our results show that the shaping time can be an important tool to adapt the pulse length and noise level to the photon flux and thereby optimize the dose efficiency of photon-counting silicon detectors. (C) The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License.Funding Agencies|Erling-Persson Family Foundation; MedTechLabs; European Unions Horizon 2020 Research and Innovation Programme [830294]; project CALIPSO plus from the EU Framework Programme for Research and Innovation HORIZON 2020 [730872]</p

Author Correction: Robust estimation of bacterial cell count from optical density

An amendment to this paper has been published and can be accessed via a link at the top of the paper.</jats:p

Robust estimation of bacterial cell count from optical density

AbstractOptical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals  &lt;1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data.</jats:p