鼻咽癌细胞双光子显微图像处理

鼻咽细胞的双光子显微图像中含有着丰富信息,借助计算机和图像处理算法可进行分析处理。图象分割是双光子显微图象处理中的一项重要技术,至今为止尚未形成一个最佳通用方法,也没有定义出双光子显微图象分割的统一标准。本文首先采用噪声干扰法进行去噪,采用低帽的变换等的数学形态学来增强鼻咽癌细胞图像,使细胞更加容易分辨,接着对几种经典边缘检测算法进行讨论比较,紧接着根据鼻咽双光子显微图像的实际特征,采取腐蚀算法求出鼻咽癌细胞边缘。然后进行区域生长定位细胞,并采用一些改进的判别分析算法和区域面积算法对鼻咽癌细胞进行阈值分割,获得较好结果。     

激光共聚焦显微技术在植物学中的应用

激光扫描共聚焦显微镜（Ｌａ－ｓｅｒ　Ｓｃａｎｎｉｎｇ　Ｃｏｎｆｏｃａｌ　Ｍｉｃｒｏｓｃｏｐｅ,ＬＳＣＭ）是近年来发展起来的一种新型高精度显微镜,它在荧光显微镜成像基础上加装了激光扫描装置,使用可激发的荧光探针对样品进行标记,利用计算机进行图像采集处理,从而可得到样品内部细微结构的荧光图像。目前此种显微技术不仅用于观察经固定的各种细胞和组织结构,而且还可对活细胞的形态、结构,离子实时动态等进行观察和定量荧光测定,以及定量图像分析。另外,该仪器还具备样品断层扫描,三维图像重建的独特功能。因此,共聚焦…     

活体细胞三维图像科学可视化方法的研究

科学可视化是指运用计算机图形学和图像处理技术,将科学计算过程中或者是计算结果的数据转换为图形或图像,在屏幕上显示出来并进行交互式处理的理论技术或方法。介绍了用反卷积荧光显微成像技术获得活体大鼠胰腺B细胞三维图像及对其进行科学可视化的主要过程和两种常用可视化算法,并运用这两种方法对所得到的三维图像进行处理以分析和研究细胞内分泌囊泡的空间分布。结果显示,当仅观察细胞三维图像的二维切片时,三维图像中的某些重要信息会被忽略,而使用科学可视化方法则可以从三维角度直观观察活体细胞内分泌囊泡的空间分布,并且可以观察到分泌囊泡的释放趋势和整体分布,从而为细胞生物学研究提供重要的信息。     

小波变换及其在医学图像处理中的应用

医学图像的好坏直接影响着医生对病情的诊断和治疗,因此利用数字图像处理等技术对医学图像进行有效的处理,已成为医学图像处理研究和开发的一大热点。小波变换是对傅里叶变换的继承和发展,在医学影像领域有着广泛的应用前景。本文介绍了二维离散小渡变换的一般形式,在图像分解与重构的基础上．系统地阐述了利用小小组变换的时频域特性与多分辨分析对医学图像进行去噪、增强以及边缘提取等深层次的处理,有效的改善图像质量。     

EBV感染及TPA处理前后人CR2转染细胞的基因差异表达谱

采用Mouse Atlas^TM cDNA Expression Arrays对EBV感染前后及TPA处理前后的人CR2转染小鼠细胞基因表达进行分析,通过Eagle EyeⅡ图像分析系统进行密度扫描以寻找差异表达基因。结果表明已初步建立EBV和TPA的转染细胞基因差异表达谱,为进一步研究二者对转染小鼠 细胞的影响奠定了良好基础,也为进一步发现转染小鼠细胞中EBV和TPA相关的信号转导通路提供线索。     

基于小波分析的医学图像的处理

医学图像的好坏直接影响着医生对病情的诊断和治疗,因此利用数字图像处理等技术对医学图像进行有效的处理,已成为医学图像处理研究和开发的一大热点.小波分析是对傅立叶变换的继承和发展,在医学影像领域有着广阔的应用前景.介绍了二维离散小波变换的一般形式,在图像分解的基础上,利用小波分析对医学图像进行去噪和增强处理,能够有效的改善图像质量,有利于医生对病情的诊断和治疗.     

高灵敏度荧光显微镜量化技术及其在光生物学研究中的应用

介绍一种用像增器接收荧光图像的高灵敏度荧光显微镜,相对于普通荧光显微镜的灵敏度提高了４×１０４倍,并用宽量程微光光亮度计对仪器的微弱成像性能进行了实验标定,得到了图像采集数据和图像发光强度的线性数量关系。高灵敏度荧光显微镜在给出细胞荧光图像的同时,可以给出图像上每一像元的发光强度和细胞平均发光强度,仪器对图像细微变化的判断能力远大于人眼直接观察图像。高灵敏度荧光显微镜可应用于研究细胞中荧光物质在细胞生理过程中的分布变化和发光强度变化。使用此仪器已得到了光敏竹红菌甲素（ＨＡ）在Ｈｅｌａ细胞（人体子宫癌细胞）中的分布图像和更为直观的三维显示图形,以及加入ＨＡ后Ｈｅｌａ细胞受到强先照射后的细胞损伤图像。     

基于Visual C#语言的DICOM标准医学图像的显示和实现

介绍DICOM3.0医学图像文件的格式和C#语言的特点,首次利用Visual C#语言对该标准的图像进行显示和处理,能够直接读取DICOM格式原始图像数据,并可批量转换成BMP等格式进行处理,此项工作可为医学图像处理研究及相关医学图像软件开发奠定基础。     

基于MIP图像拼接系统的大型古生物化石数字化应用研究北大核心CSCD

在大型古生物化石数字化过程中,为了充分展示化石的细节信息,往往需要拍摄大量的图像。为了实现大型古生物化石数字化数据的完整性,需要对这些大量的图像进行精密的图像拼接处理。基于这种应用需求的前提下,本文在自主研发的Mosaic of Image Program(MIP)图像拼接系统的基础上,对高精度的相机检校、畸变检校及改正和拼接缝的保真处理等方面进行研究,形成系统的古生物化石彩色合成影像数字化流程。在宜州化石馆的实际处理中,完成了杨氏锦州龙、蜥脚类恐龙、孔子鸟等大型古生物化石的数字化,几何失真小于0.36mm(畸变矫正精度优于1像元,拼接精度优于2像元,像片分辨率0.12mm)。同时采用基于SIFT的自动辐射归一化处理算法对拼接影像进行辐射均衡处理,矫正拼接影像辐射亮度的不均衡。     

激光扫描共聚焦显微术在生物医学中的研究

激光扫描共聚焦显微技术的发展,为形态学、分子细胞生物学、神经科学、肿瘤医学、药理学、遗传学等领域的研究提供了新的先进研究方法和手段。本文对激光扫描共聚焦显微术在粘附细胞分选,动态荧光分析测定,显微细胞外科和光陷阱技术,荧光光漂白恢复技术以及三维图像重建功能等方面的研究进展进行了综述。     

Image analysis method for the rapid counting of Saccharomyces cerevisiae cells

An image analysis system which incorporates a microscope, video camera, monitor, and Apple computer and which uses image area to count Saccharomyces cerevisiae cells is described and evaluated. Yeast cell suspensions of densities of up to 100 X 10(6) cells per ml can be counted when viewed in the counting chamber of a hemacytometer. The yeast image area measured depends upon the light intensity used to illuminate the yeast cells, the sharpness of the image focused on the monitor, and the grey level selected when scanning the digitized image on the monitor of the Apple computer, all of which can be controlled. The image area also depends upon the yeast strain and medium in which the culture is grown, but it is not affected by the concentration of sugar or ethanol in which the yeast cells are suspended. Yeast growth measured by image analysis can be calibrated to give results similar to those obtained with hemacytometer counting. Yeast cells can be counted in the presence of high cell densities of bacteria by adjusting the grey level at which the digitized image is scanned.     

Image analysis method for the rapid counting of Saccharomyces cerevisiae cells.

An image analysis system which incorporates a microscope, video camera, monitor, and Apple computer and which uses image area to count Saccharomyces cerevisiae cells is described and evaluated. Yeast cell suspensions of densities of up to 100 X 10(6) cells per ml can be counted when viewed in the counting chamber of a hemacytometer. The yeast image area measured depends upon the light intensity used to illuminate the yeast cells, the sharpness of the image focused on the monitor, and the grey level selected when scanning the digitized image on the monitor of the Apple computer, all of which can be controlled. The image area also depends upon the yeast strain and medium in which the culture is grown, but it is not affected by the concentration of sugar or ethanol in which the yeast cells are suspended. Yeast growth measured by image analysis can be calibrated to give results similar to those obtained with hemacytometer counting. Yeast cells can be counted in the presence of high cell densities of bacteria by adjusting the grey level at which the digitized image is scanned.     

Population estimation of human embryonic stem cell cultures

Traditionally, the population of human embryonic stem cell (hESC) culture is estimated through haemacytometer counts, which include harvesting the cells and manually analyzing a fraction of an entire population. Obviously, through this highly invasive method, it is not possible to preserve any spatial information on the cell population. The goal of this study is to identify a fast and consistent method for in situ automated hESC population estimation to quantitatively estimate the cell growth. Therefore, cell cultures were fixed, stained, and their nuclei imaged through high‐resolution microscopy, and the images were processed with different image analysis techniques. The proposed method first identifies signal and background by computing an image specific threshold for image segmentation. By applying a morphological operator (watershed), we split most physically overlapping nuclei, leading to a pixel area distribution of isolated signal areas on the image. On the basis of this distribution, we derive a nucleus area model, describing the distribution of the area of cell debris, single nuclei, and small groups of connected nuclei. Through the model, we can give a quantitative estimation of the population. The focus of this study is on low‐density human embryonic stem cell populations; hence cultures were measured at days 2–3 after seeding. Compared with manual cell counts, the automatic method achieved higher accuracy with <6% error. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Application of image analysis on monolayer cultures of rat hepatocytes: assessment of a chemically inducible G0-G1 cell cycle phase shift.

The phenobarbital induced shift from G0 to G1 cell cycle phases was analyzed in freshly isolated cultured rat hepatocytes by image analysis. Nuclei in situ in monolayers or in an isolated state were stained with quinacrine dihydrochloride. Fluorescence intensity and fluorescence area were recorded in controls and after treatment with phenobarbital (1.5 or 3 mM, 48 h). Reproducible measurements were obtained with the aid of an elaborate background correction and image enhancement procedure and by the construction of individual measuring masks for each nucleus. A complete statistical analysis revealed that in both preparations (isolated nuclei and monolayer cultures, treated and untreated), individual ploidy classes were distinguishable by fluorescence area measurements. Within each ploidy class, the area is modified by the cell density: with increasing cell density the area occupied by a single cell decreases. After phenobarbital treatment, a decrease in size, due to the higher cell density after the mitotic stimulus of the test compound and a decrease in total fluorescence, due to the G0-G1 cell cycle phase shift was recorded. In monolayer cultures, but not in isolated nuclei, two populations of nuclei were discernible suggesting two cell populations, one responding to treatment and one refractive.     

Quantitation of cell area on glass and fibronectin-coated surfaces by digital image analysis.

By using digital image processing and analysis, two procedures were developed to rapidly measure the projected area of a field of adherent 3T3 fibroblasts without staining of cell borders. The cell area of newly attached and rounded cells with well-resolved borders was obtained by a gray value thresholding procedure. For cells that had undergone an appreciable degree of spreading, cell boundaries were less distinct and a nonlinear spatial Sobel filter was used, followed by thresholding. For both procedures, linear relations were observed between cell areas obtained from image analysis and cell areas obtained by tracing. The areas of a population of traced cells were not statistically different from the area distribution obtained by using the standard curves for the processed images. Uncertainty in the estimated mean area depended only upon the number of cells examined. Approximate numbers of cells required to obtain estimates of the mean are calculated. As an application of these procedures, cell areas were measured for 3T3 cells attached to glass and fibronectin-coated surfaces and were found to be significantly larger for cells spreading on fibronectin-coated glass than on glass alone. Increased cell area during spreading on fibronectin-coated surfaces was proportional to increased cell adhesivity after exposure to a shear stress of 58 dyn/cm2.     

Simple method for quantification of fast plasma membrane movements.

We present a method for the quantification of the fast plasma membrane movements that are involved in ruffling, blebbing, fast shape change, and fast translocation. The method is based on the Kontron Vidas image analysis computer program. Video images from cells viewed through an inverted microscope were transmitted to the computer. The procedure was as follows: 4 consecutive video images were averaged (image 1); 28 s later a second set of 4 video images was averaged (image 2); image 2 was subtracted from image 1 and the grey level of each pixel of the resulting image was increased with 128 grey level units, resulting in the subtraction image, showing a uniform grey background speckled with brighter and darker spots corresponding to areas of movement. These spots were discriminated and turned into white objects against a black background. Interactive editing was used to delete artefacts that resulted from floating debris. The total area of the discriminated objects was measured, and the parameter motile area in micron2 per cell was calculated. We have applied our method to the study of motility induced in epithelial cell lines by the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate and by epidermal growth factor.     

Quantitative oral exfoliative cytology. Effect of alcohol on normal buccal mucosa

OBJECTIVE: To assess the effect of chronic alcohol intake on the DNA distribution and cell area of normal oral mucosal cells. STUDY DESIGN: Smears were taken from clinically normal buccal mucosa of 50 patients attending an alcohol-problem service (i.e., chronic alcohol use) and average alcohol units per week recorded. DNA distribution histograms and total cell area values were then compared to those obtained from smears taken from a control group (which included social drinkers) of patients attending for routine dental treatment. Nuclear DNA content was assessed on 100 randomly selected, Feulgen-stained nuclei using a Seescan TV image analysis system, and total cell area was assessed on 50 Papanicolaou-stained cells using the Vids V image analysis system. RESULTS: The DNA distribution histograms were essentially diploid in appearance for the alcohol group, although there was an increase in nuclear DNA content in the occasional nucleus. A highly significant reduction in total cell area was found for the alcohol group when compared to the controls. CONCLUSION: The chronic ingestion of alcohol is associated with a reduction in total cell area but appears to have little effect on nuclear DNA content. Our previous research using the same technique showed that oral cancers are frequently nondiploid. Thus, a nondiploid DNA distribution histogram for smears taken from a clinically suspicious lesion in someone who consumes excessive amounts of alcohol is unlikely to be due to alcohol use alone and should indicate biopsy.     

A "two-objective, one-area" procedure in absorption microphotometry and its application using an inverted microscope

A 'two-objective, one-area' method and related equations are suggested to measure absorbance of microscopic stained objects. In such work, the measuring field invariably includes an image of the object and some clear area surrounding the image. The total intensity in the two areas is measured photometrically, using two different objectives, and substituted in the equation for absorbance. The equation is independent of the term representing intensity from the clear area and hence the error in the measurement of absorbance is reduced. The limitations of the 'two-objective, one-area' method are discussed and its pragmatic operation described with an experimental setup involving an inverted microscope. The method permits measurement of intensity in a part of a stained cell while the rest of the cell remains in the field of view. The method is applied to measure absorbance in Giemsa stained ascites cells and Feulgen stained liver and Human Amnion cells.     

Image-based high-throughput quantification of cellular fat accumulation

A number of biochemical methods are available for measuring fat accumulation in cell culture. The authors report a simple image-based method for measuring fat accumulation in adipocytes using a combination of high-throughput brightfield microscopy and image analysis, which was validated biochemically using Oil-Red-O. The quickest and most accurate method of analysis was one based on thresholding brightfield images and determining the area of fat droplets per image. Thus, the authors have developed a simple high-throughput, label-free method for measuring fat accumulation that is applicable to any cell or tissue type where fat droplets are visible under light microscopy.