The analysis of DNA and protein in a single cell by Hadamard transform microscope image

Abstract

The Hadamard transform microscope image analysis was applied to analyze the DNA and protein in a single cell. By this method, the fluorescence emission spectrum, the fluorescence image and the fluorescence intensity of a single cell can be acquired which was tagged by a fluorescence probe. The potential application value of this system is shown for cytobiological aspects.Theory of the instrument. As shown in Fig. 1, the light signal captured by the objective can directly enter and be focussed on the focus-plane of the CCD camera when the mirror (M) is moved away from the optical path, then the image of analyzed sample can be Fresenius Fresenius' Journal of Fig. 1. Schematic diagram of the instrument of Hadamard transform microscope fluorescence image. P: dichroic beamsplitter, L 1 , L 2 , L 3 , L 4 , L 5 , L 6 : lens, M: mirror

Analysis of DNA in Single Pollen Cell by Hadamard Transform Microscopic Fluorescence Imaging

“…The theory and structure of the instrument were described in detail in our previous studies (13,14). The three-dimensional image of a cell sample can be obtained as an image of 15 ϫ 17 pixels (two spatial dimensions, one spectrum dimension).…”

“…It has three advantages: (1) Fellgett's advantage, by which an improvement in the signal-to-noise ratio can be obtained; (2) the power distribution advantage; and (3) the source-encoded and signal-encoded image (10)(11)(12). In order to develop a new image analysis method for cellular quantitative analysis, in our previous studies, the technique of Hadamard transform multiplexed imaging was coupled with microscopy and a novel method for microscopic analysis was developed (13,14).…”

Measurements of the DNA Content in a Breast Tumor Cell Based on the Hadamard Transform Microscopic Fluorescence Image

“…[19][20][21] Our previous studies have demonstrated the principles of Hadamard transform microscopic fluorescence imaging. [22][23][24] In this work, a new instrument, Hadamard transform (HT) microscopic fluorescence image analysis system, which was developed in our laboratory, was applied to analyze the nuclear DNA content in breast tumor cells with Acridine Orange (AO) being used as a fluorescence probe for cellular DNA. The results show a high concordance with a pathological diagnosis, which indicates that the technique is potentially applicable to diagnostic and prognostic studies on tumors.…”

Quantitative DNA Imaging in Breast Tumor Cells by a Hadamard Transform Fluorescence Imaging Microscope

“…13 The application studies of our previous system to single-cell quantitative analysis indicate that 2D fluorescence microscopic imaging by Hadamard multiplexing is suitable for biological samples. [14][15][16] Little has been reported recently on HT spectral imaging for fluorescence microscopy, to our knowledge, except our previous reports for 15 × 17 pixel imaging. [13][14][15][16] Liquid crystal spatial light modulator (SLM) has been applied for Hadamard 2D spectral imaging with a charge-coupled-device (CCD) camera in a fluorescence microscope, 17,18 but the mathematical operation was prolix because a total of 511 768 × 512 pixel images were collected and the total acquisition time was quite long.…”

“…[14][15][16] Little has been reported recently on HT spectral imaging for fluorescence microscopy, to our knowledge, except our previous reports for 15 × 17 pixel imaging. [13][14][15][16] Liquid crystal spatial light modulator (SLM) has been applied for Hadamard 2D spectral imaging with a charge-coupled-device (CCD) camera in a fluorescence microscope, 17,18 but the mathematical operation was prolix because a total of 511 768 × 512 pixel images were collected and the total acquisition time was quite long. Recently in our group, a HT microscopic imaging system which could be used to generate 511 × 512 pixel format images for small samples was developed by using a one-dimensional movable mask and a monochromator with a simplified optical path.…”