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Instrument for Hadamard transform three-dimensional fluorescence microscope image analysis 总被引:2,自引:0,他引:2
Guanquan Chen Erwen Mei Wenfang Gu Xiaobin Zeng Yun'e Zeng 《Analytica chimica acta》1995,300(1-3):261-267
An instrument combining fluorescence microscopy with Hadamard transform multiplexed imaging was designed by which a three-dimensional Hadamard transform fluorescence microscopic cell image was obtained. The image can provide useful information including, simultaneously, the apparent dimensions and the shape of the analytical sample, the content and the distribution of some species in it. 相似文献
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With a 511-slit one-dimensional (1D) Hadamard mask and a highly sensitive linear charge-coupled device (CCD), spatial multiplexing is performed and a programmable Hadamard transform (HT) microscopic fluorescence imaging system was developed. The system can generate 511×512 pixel format images for small samples. Sensitivity, signal to noise ratio, imaging speed and spatial resolution of this system were discussed. The results show that the system can be applied for single-cell imaging sensitively in a short time. Spatial resolution up to 0.24 μm/pixel, which is close to the resolution limit of the conventional optical microscope, has been obtained under oil lens. The weak native fluorescence imaging for pollen cells can be realized within 1 min. The system has been applied for multi-parameter evaluation of tumor malignancy based on nuclear DNA ploidy measurements for one breast tumor specimen. The result indicates that the system has good application prospect in cell biology and medicine. 相似文献
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使用一维机械模板的高分辨阿达玛变换显微图像探测技术研究 总被引:2,自引:1,他引:1
阿达玛变换(HT)是一种类似于傅里叶变换(FT)的光谱调制技术,具有多通道同时检测和多通道成像能力等优点,但两者的数学模型、对光信号的调制方法和调制手段都不一样。由于HT仅涉及四则运算,而FT涉及较为复杂的三角函数和复数运算,所以HT的解码速度快于FT。在成像技术方面,HT具有直接成像的能力,而FT只能对通过其它方式获取的图像进行加工处理。 相似文献
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This paper presents a novel Hadamard transform (HT) fluorescence imaging microscope by combining multiplexed imaging technique with a conventional upright fluorescence microscope for single-cell imaging and quantitative cellular analysis. The HT imaging microscope can provide 511 x 512-pixel single-cell image with high sensitivity within 21 s. In this study, the high potential value of the microscope in biomedical analysis has been demonstrated by using it to evaluate the malignancy degree of thirty cases of human breast tumors based on the measurements of cellular DNA contents, with conclusions highly accordant with pathological diagnosis. The results show that the HT microscope has the ability to analyze very small specimens and the capability of detecting very high ploidy cells, which are advantages over flow cytometry. The microscope was also successfully applied to cellular morphological analysis, and it was demonstrated that a significant linear relationship exists between tumor nuclear DNA contents and the nuclear area, and malignant and benign tumors are significantly different in both DNA contents and nuclear area. The reliability of the HT microscope in cellular DNA measurements was also investigated. 相似文献
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提出了一种改进的阿达玛变换(HT)显微荧光图像分析系统,以单细胞试样分析为基础,分别对系统的分辨率和解码后的图像恢复过程进行了讨论.结果表明,该系统可应用于单细胞形态分析和定量分析.图像在x和y方向的像素分辨率相同,并达到了同一成像物镜下的空间分辨率水平,因此在获取微米级单细胞试样的微弱荧光信号的二维图像时,系统的成像能力较好,可用于单细胞形态分析.对花粉细胞的荧光衰退过程的定量分析结果表明,对不同HT图像提供的同一系列试样的定量数据进行比较时,必须对所有该系列试样的图像恢复过程进行归一化处理. 相似文献
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阿达玛变换(Hadamard transform, HT)是一种类似于傅里叶变换的光谱调制技术, 具有多通道同时检测和多通道成像能力. 实现高分辨HT成像的关键在于阿达玛模板的制作, 阿达玛模板有两种, 即移动式机械编码模板(Movable mechanical mask)和固定式光电模板(Stationary electro-optic mask). 在实际成像方面, 移动模板和固定模板各有优缺点: 前者一般用石英玻璃制作, 对光信号不会因模板吸收而导致信号损失, 因此数据很可靠, 而且模板的制作也较为容易, 但由于采用步进电机驱动而容易导致机械故障, 难以实现快速编码; 后者无移动部件, 无机械故障, 因此系统比较紧凑, 但由于它是由液晶材料制成的(可导致信号损失), 从而限制了其在某些光谱区域的使用. 此外, 它对系统的软件设计要求比前者高, 实现高分辨成像更加困难. 正是由于上述原因, 实现快速、高分辨HT成像具有一定难度, 最近有关HT成像技术的报道极少. 相似文献
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阿达玛变换显微图象分析系统在乳腺肿瘤细胞DNA倍性分析中的应用研究 总被引:2,自引:0,他引:2
以正常人外周静脉血淋巴细胞为标准二倍体细胞,以吖啶橙为细胞DNA荧光探针,用阿达玛变换显微图象分析仪测定了六例乳腺肿瘤的细胞DNA含量(倍性),分析结果与病例学诊断结论吻合,表明该仪器可望用于乳腺癌的诊断和预后研究.研究结果还表明:乳腺肿瘤细胞DNA倍性与核面积(象素)呈显著正相关. 相似文献
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Recent applications of scanning electrochemical microscopy (SECM) to studies of single biological cells are reviewed. This
scanning probe microscopic technique allows the imaging of an individual cell on the basis of not only its surface topography
but also such cellular activities as photosynthesis, respiration, electron transfer, single vesicular exocytosis and membrane
transport. The operational principles of SECM are also introduced in the context of these biological applications. Recent
progress in techniques for high-resolution SECM imaging are also reviewed. Future directions, such as single-channel detection
by SECM, high-resolution imaging with nanometer-sized probes, and combined SECM techniques for multidimensional imaging are
also discussed. 相似文献
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Johnson RD Navratil M Poe BG Xiong G Olson KJ Ahmadzadeh H Andreyev D Duffy CF Arriaga EA 《Analytical and bioanalytical chemistry》2007,387(1):107-118
Bulk studies are not suitable to describe and study cell-to-cell variation, which is of high importance in biological processes
such as embryogenesis, tissue differentiation, and disease. Previously, capillary electrophoresis with laser-induced fluorescence
detection (CE-LIF) was used to measure the properties of organelles isolated from millions of cells. As such, these bulk measurements
reported average properties for the organelles of cell populations. Similar measurements for organelles released from single
cells would be highly relevant to describe the subcellular variations among cells. Toward this goal, here we introduce an
approach to analyze the mitochondria released from single mammalian cells. Osteosarcoma 143B cells are labeled with either
the fluorescent mitochondrion-specific 10-N-nonyl acridine orange (NAO) or via expression of the fluorescent protein DsRed2. Subsequently, a single cell is introduced
into the CE-LIF capillary where the organelles are released by a combined treatment of digitonin and trypsin. After this treatment,
an electric field is applied and the released organelles electromigrate toward the LIF detector. From an electropherogram,
the number of detected events per cell, their individual electrophoretic mobilities, and their individual fluorescence intensities
are calculated. The results obtained from DsRed2 labeling, which is retained in intact mitochondria, and NAO labeling, which
labels all mitochondria, are the basis for discussion of the strengths and limitations of this single-cell approach.
Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users 相似文献
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Fourier transform infrared (FTIR) spectroscopic imaging is a relatively new method that has received great attention as a
new field of analytical chemistry. The greatest benefit of this technique lies in the high molecular sensitivity combined
with a spatial resolution down to a few micrometers. Another advantage is the ability to probe samples under native conditions,
which allows new insights into samples without the need for fixation, stains, or an additional marker. Advances in instrumentation
have made FTIR spectroscopic imaging the tool of choice for an increasing number of applications. The main applications are
in the bioanalytical chemistry of cells and tissue, polymers, and recently as well as in homeland security. This report gives
a short overview of current developments and recent applications.
Figure FTIR image of a polymer blend reveals the chemical composition. Online Abstract Figure (365 KB). 相似文献
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Subhadeep Banerjee Apeksha Ashok Phadte Anjan Chattopadhyay Joseph R.D. Fernandes Akanksha Vinod Savardekar Arnab Banerjee Keisham Sarjit Singh 《Tetrahedron letters》2018,59(42):3787-3791
Aza- and oxa-dihydrotetracenes were prepared in one step from 1,4-dicyanodibenzodioxins in good yields. These molecules represent the first examples of heterocyclic dihydrotetracenes with push-pull character. Aza-dihydrotetracene 18H showed a strong red-shifted absorbance maxima at 472?nm, nearly equal in intensity to the parent band at 260?nm, in polar DMSO medium. The fluorescence emission spectrum of aza-dihydrotetracene 18H had an emission maximum at 525?nm with a broad band stretched out as far as 650?nm. The fluorescence emission quantum yield was almost as strong as the known fluorescent standard 1,3-diphenylisobenzofuran. The aza-dihydrotetracenes exhibited in vitro cytotoxicity against the HeLa cell line and were non-toxic against the normal HaCaT cell line. The fluorescence emerging from HeLa cells incubated with aza-dihydrotetracene 18H was very strong and helped detect the cells microscopically using appropriate filters. 相似文献
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Short oxidized multi-walled carbon nanotubes (CNT) were derivatized with fluorescein isothiocyanate (FITC). Capillary electrophoresis
coupled with laser-induced fluorescence (CE–LIF) was then used to separate and detect the fluorescently labeled carbon-nanotube
probes (CNTP) in multidrug-resistant cells (K562A) and the parent cells (K562S). Greater expression of P-glycoprotein in K562A
cells than in K562S cells was confirmed by use of anti-P-glycoprotein antibody and flow-cytometric analysis. Analyses of CNTP
in both cell lines using both CE–LIF and flow cytometry showed that CNTP could traverse the cellular membrane without being
pumped out by P-glycoprotein. The CNTP distributed in both cell lines was analyzed at the single cell level and the results
were compared with those from analysis of ten cells and of the lysate from bulk cells. The results revealed the CE–LIF method
could be used for quantitative analysis of CNT in single cells in studies of drug delivery and multidrug resistance.
相似文献
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Phosphatase plays a vital important role in many biological functions due to the dephosphorylation serves varied roles in cellular regulation and signaling.Among the family of phosphatase,alkaline phosphatase(ALP) could act as crucial prognostic indicators for many diseases such as bone diseases and cancer.However,the detection of ALP is mainly limited to in vitro colorimetric method in clinic.Therefore,huge efforts have been paid on the fluorescence imaging that provides a reliable method to detect the real-time and in vivo changes of the level of ALP.In this review,we summarize recent advances in fluorescence imaging of phosphatase,mainly focused on ALP.The imaging probes of phosphatase are mainly classified according to their luminescence mechanisms.In the end,we assessed the challenges and future prospects of phosphatase probes. 相似文献
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We developed a simple fluorescence microscopy for acquisition of high-resolution images of single quantum dots (QDs) labeled to biomolecules on apical plasma membrane, in cell interior and on basal plasma membrane of living cells. The method was a combination of total internal reflection fluorescence microscopy (TIRFM) at apical cell surface and intracellular microscopy coupled with focusing objective. Insulin conjugated to single QD (insulin-QD) was chosen as the model system. In order to bind insulin-QDs to insulin receptors on the plasma membrane through the interaction between insulin and its receptor, as well as internalize them, the cells attached on a coverslip were incubated with biotinylated insulin and QD-streptavidin conjugate at 37 °C. Next, fluorescent molecules in the cells were photobleached by illuminating the cells using a 100-W mercury lamp with the wavelengths from 460 to 490 nm. Then, the incident angle of a laser beam was adjusted to produce total internal reflection at the apical surface of a single cell. In this case, the insulin-QDs in the whole cell were excited, and the fluorescent molecules outside the cell were not illuminated. Finally, the images of single insulin-QDs on the apical plasma membrane, in the cell interior and on the basal plasma membrane of the cell were taken by focusing the objective to different positions, respectively. The resolution and contrast of the fluorescent spots in the images were much higher than those obtained by using epi-fluorescence microscopy and comparable to those obtained by using the conventional TIRFM. The method improved the image acquisition speed for the images on the apical and basal plasma membrane using the conventional TIRFM, and could acquire the high-resolution images in the cell interior quickly. 相似文献
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基于1,8-萘酰亚胺衍生物,构建了一种检测半胱氨酸(Cys)的新型荧光探针TPFC-Acryloyl。光谱研究表明该探针能有效识别Cys且能够在1min内实现快速响应。探针对Cys的检测表现出高选择性,检测限为2.13μmol/L。经荧光光谱和质谱实验确证其检测机理为:Cys与TPFC-Acryloyl分子中的丙烯酸酯发生共轭加成-环化反应,进而羟基裸露的同时释放出黄色荧光。细胞毒性测试表明探针TPFC-Acryloyl的细胞毒性低。此外,该探针还被成功应用于活细胞和秀丽隐杆线虫中Cys的荧光成像。 相似文献