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

Thicker Retinal Nerve Fiber Layer with Age among Schoolchildren: The Hong Kong Children Eye Study

This study aims to investigate the effect of age on the peripapillary retinal nerve fiber layer (p-RNFL) thickness among schoolchildren. A total of 4034 children aged 6–8 years old received comprehensive ophthalmological examinations. p-RNFL thickness was measured from a circular scan (⌀ = 3.4 mm) captured using spectral-domain optical coherence tomography (SD-OCT). Associations between p-RNFL thickness with ocular and systemic factors were determined by multivariate linear regression after adjusting potential confounders using generalized estimating equations (GEE). The mean global p-RNFL thickness was 106.60 ± 9.41 μm (range: 72 to 171 μm) in the right eyes, 105.99 ± 9.30 μm (range: 76 to 163 μm) in the left eyes, and 106.29 ± 9.36 μm (range: 72 to 171 μm) across both eyes. Age was positively correlated with p-RNFL after adjusting for axial length (AL) and confounding factors (β = 0.509; p = 0.001). Upon multivariable analysis, AL was positively associated with temporal p-RNFL thickness (β = 3.186, p < 0.001) but negatively with non-temporal p-RNFL thickness (β = (10.003, −2.294), p < 0.001). Sectoral p-RNFL was the thickest in the inferior temporal region (155.12 ± 19.42 μm, range 68 to 271 μm), followed by the superior temporal region (154.67 ± 19.99 μm, range 32 to 177 μm). To conclude, p-RNFL increased significantly with older age among children 6 to 8 years old in a converse trend compared to adults. Our results provide a reference for interpreting OCT information in children and suggest that stable p-RNFL thickness may not indicate a stable disease status in pediatric patients due to the age effects

Erratum to: Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition) (Autophagy, 12, 1, 1-222, 10.1080/15548627.2015.1100356

non present