Gutiérrez-Medina Braulio, Sánchez Miranda Manuel de Jesús
Division of Advanced Materials, Instituto Potosino de Investigación Científica y Tecnológica (IPICYT), San Luis Potosí, Mexico; Division of Molecular Biology, Instituto Potosino de Investigación Científica y Tecnológica (IPICYT), San Luis Potosí, Mexico.
Division of Advanced Materials, Instituto Potosino de Investigación Científica y Tecnológica (IPICYT), San Luis Potosí, Mexico.
Biophys J. 2017 Nov 7;113(9):1916-1919. doi: 10.1016/j.bpj.2017.09.002. Epub 2017 Oct 4.
Bright field (BF) optical microscopy is regarded as a poor method to observe unstained biological samples due to intrinsic low image contrast. We introduce quantitative image restoration in bright field (QRBF), a digital image processing method that restores out-of-focus BF images of unstained cells. Our procedure is based on deconvolution, using a point spread function modeled from theory. By comparing with reference images of bacteria observed in fluorescence, we show that QRBF faithfully recovers shape and enables quantify size of individual cells, even from a single input image. We applied QRBF in a high-throughput image cytometer to assess shape changes in Escherichia coli during hyperosmotic shock, finding size heterogeneity. We demonstrate that QRBF is also applicable to eukaryotic cells (yeast). Altogether, digital restoration emerges as a straightforward alternative to methods designed to generate contrast in BF imaging for quantitative analysis.
明场（BF）光学显微镜由于其固有的低图像对比度，被认为是观察未染色生物样本的较差方法。我们引入了明场定量图像恢复（QRBF），这是一种数字图像处理方法，可恢复未染色细胞的离焦明场图像。我们的程序基于反卷积，使用从理论建模的点扩散函数。通过与荧光观察到的细菌参考图像进行比较，我们表明QRBF能够忠实地恢复形状，并能够量化单个细胞的大小，即使是从单个输入图像中也能做到。我们将QRBF应用于高通量图像细胞仪，以评估大肠杆菌在高渗休克期间的形状变化，发现了大小异质性。我们证明QRBF也适用于真核细胞（酵母）。总之，数字恢复成为一种直接的替代方法，可替代旨在在明场成像中产生对比度以进行定量分析的方法。
