High-resolution Hadamard transform microscope fluorescence imaging: quantifying the DNA content in single cells.
This study presents a novel miniaturized Hadamard transform fluorescence imaging microscope, by combining a conventional fluorescence microscope with Hadamard transform multiplexing encoding using a one-dimensional movable mask to realize spatial resolution and a linear CCD for multichannel detection. The microscope can provide high-resolution automatically-generated 0-255 gray level HT images for morphological analysis and visualization of a single cell, and normalized HT images for cellular quantitative measurements. The microscope's imaging capability was applied to measure the DNA content in human lymphocyte, chicken erythrocyte and eel erythrocyte, and a comparative study was performed. The results show that the calibrated DNA content in a chicken erythrocyte is 2.32 pg when human lymphocyte is used as the standard, and eel erythrocyte may be a potentially reliable and novel standard for determining DNA contents in other species because it has a stable DNA value of 2.06 pg, with a CV of 4.3% when 20 eel erythrocytes are measured. The results also demonstrate that the HT imaging microscope should be valuable in the fields of medicine and cell biology.
Tang
Hongwu
H
Center of Analysis and Measurements, College of Chemistry and Molecular Science, Wuhan University, Wuhan, 430072, P. R. China. hwtang@whu.edu.cn
Luo
Meina
M
Xiong
Yongpan
Y
Chen
Guanquan
G
D004247:DNA / Q000032:analysis*; D008856:Microscopy, Fluorescence / Q000379:methods*; D015999:Multivariate Analysis
D004247:DNA