TY - JOUR
T1 - Quantitative Image Restoration in Bright Field Optical Microscopy
AU - Gutiérrez-Medina, Braulio
AU - Sánchez Miranda, Manuel de Jesús
N1 - Publisher Copyright:
© 2017 Biophysical Society
PY - 2017/11/7
Y1 - 2017/11/7
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85030483157&partnerID=8YFLogxK
U2 - 10.1016/j.bpj.2017.09.002
DO - 10.1016/j.bpj.2017.09.002
M3 - Artículo
C2 - 28988026
AN - SCOPUS:85030483157
SN - 0006-3495
VL - 113
SP - 1916
EP - 1919
JO - Biophysical Journal
JF - Biophysical Journal
IS - 9
ER -