Analytical Cellular Pathology - Volume 36, issue 1-2

Authors: Cohen, Stanley

Article Type: Other

DOI: 10.3233/ACP-2013-0079<

Citation: Analytical Cellular Pathology, vol. 36, no. 1-2, pp. 1-2, 2013

Authors: Fritzky, Luke | Lagunoff, David

Article Type: Review Article

Abstract: It requires a good deal of will power to resist hyperbole in considering the advances that have been achieved in fluorescence microscopy in the last 25 years. Our effort has been to survey the modalities of microscopic fluorescence imaging available to cell biologists and perhaps useful for diagnostic pathologists. The gamut extends from established confocal laser scanning through multiphoton and TIRF to the emerging technologies of super-resolution microscopy that breech the Abbé limit of resolution. Also considered are the recent innovations in structured and light sheet illumination, the use of FRET and molecular beacons that exploit specific characteristics of designer …fluorescent proteins, fluorescence speckles, and second harmonic generation for native anisometric structures like collagen, microtubules and sarcomeres. Show more

Keywords: Fluorescence microscopy, confocal microscopy, multiphoton microscopy, total internal reflectance microscopy (TIRFM), lateral sheet illumination microscopy, deconvolution, stimulated emission depletion microscopy (STED), reversible saturable/switchable optically linear fluorescence transition microscopy (RESOLFT), photoactivation localization microscopy (PALM), stochastic optical restoration microscopy (STORM), structured illumination microscopy (SIM), super-resolution optical fluctuation imaging (SOFI)

DOI: 10.3233/ACP-120071

Citation: Analytical Cellular Pathology, vol. 36, no. 1-2, pp. 5-17, 2013

Authors: Dezhampanah, Hamid | Bordbar, Abdol-Khalegh | Khodadusdt, Yadolahe

Article Type: Research Article

Abstract: The interaction of a water-soluble cationic porphyrin, Cobalt(III) 5, 10, 15, 20-tetrakis (1-methylpyridinium-4-yl) porphyrin [Co(III)TMPyP], with bovine serum albumin (BSA) has been studied in 1 mM phosphate buffer pH 7.0 containing 5 mM NaCl by UV-vis absorption, resonance light scattering (RLS) and fluorescence spectroscopies at 25°C. The results of RLS studies represent no aggregate formation of porphyrin in the surface of BSA and low tendency of this porphyrin for aggregate formation. The binding of porphyrin complex to BSA quenches fluorescence emission of BSA via a dynamic mechanism and the quenching process obeys a linear Stern-Volmer relationship. The values of Stern-Volmer …constants, KSV , was determined nearly 105 M−1 , that depend on BSA concentration. The average aggregation number of BSA calculated from the analysis of fluorescence quenching data indicates that absence of any porphyrin induced aggregation of BSA due to its interaction with porphyrin complex. The binding of Co(III) TMPyP had no obvious effect on the molecular conformation of the protein. Electrostatic force played an important role in the binding due to the opposite charges on porphyrin and the protein. Show more

Keywords: Porphyrin, serum albumin binding, fluorescence quenching, optical absorption

DOI: 10.3233/ACP-130074

Citation: Analytical Cellular Pathology, vol. 36, no. 1-2, pp. 21-26, 2013

Authors: Zhang, Tieqiao | Osborn, Samantha | Brandow, Chloe | Dwyre, Denis | Green, Ralph | Lane, Stephen | Wachsmann-Hogiu, Sebastian

Article Type: Research Article

Abstract: Structured illumination fluorescence microscopy utilizes interfering light and the moiré effect to enhance spatial resolution to about a half of that of conventional light microscopy, i.e. approximately 90 nm. In addition to the enhancement in the x and y directions, it also allows enhancement of resolution in the z- direction by the same factor of two (to approximately 220 nm), making it a powerful tool for 3-D morphology studies of fluorescently labeled cells or thin tissue sections. In this report, we applied this technique to several types of blood cells that are commonly seen in hematopathology. Compared with standard brightfield …and ordinary fluorescence microscopy images, the 3-D morphology results clearly reveal the morphological features of different types of normal blood cells. We have also used this technique to evaluate morphologies of abnormal erythrocytes and compare them with those recorded on normal cells. The results give a very intuitive presentation of morphological structures of erythrocytes with great details. This research illustrates the potential of this technique to be used in hematology and cyto-pathology studies aimed at identifying nanometer-sized features that cannot be distinguished otherwise with conventional optical microscopy. Show more

Keywords: Super-resolution microscopy, structured illumination, blood cells

DOI: 10.3233/ACP-130075

Citation: Analytical Cellular Pathology, vol. 36, no. 1-2, pp. 27-35, 2013

Authors: Qiu, Yuchen | Chen, Xiaodong | Li, Yuhua | Chen, Wei R. | Zheng, Bin | Li, Shibo | Liu, Hong

Article Type: Research Article

Abstract: BACKGROUND: Auto-focusing is an important operation in high throughput imaging scanning. Although many auto-focusing methods have been developed and tested for a variety of imaging modalities, few investigations have been performed on the selection of an optimal auto-focusing method that is suitable for the pathological metaphase chromosome analysis under a high resolution scanning microscopic system. OBJECTIVE: The purpose of this study is to investigate and identify an optimal auto-focusing method for the pathological metaphase chromosome analysis. METHODS: In this study, five auto-focusing methods were applied and tested using metaphase chromosome images acquired from bone marrow and blood specimens. These methods …were assessed by measuring a number of indices including execution time, accuracy, number of false maxima, and full width at half maximum (FWHM). RESULTS: For the specific condition investigated in this study, the results showed that the Brenner gradient and threshold pixel counting methods were the optimal methods for acquiring high quality metaphase chromosome images from the bone marrow and blood specimens, respectively. CONCLUSIONS: Selecting an optimal auto-focusing method depends on the specific clinical tasks. This study also provides useful information for the design and implementation of the high throughput microscopic image scanning systems in the future digital pathology. Show more

Keywords: Metaphase chromosome, autofocus technique

DOI: 10.3233/ACP-130077

Citation: Analytical Cellular Pathology, vol. 36, no. 1-2, pp. 37-44, 2013

Authors: Ren, Liqiang | Li, Zheng | Li, Yuhua | Zheng, Bin | Li, Shibo | Chen, Xiaodong | Liu, Hong

Article Type: Research Article

Abstract: BACKGROUND: Optimizing operational parameters of the digital microscope system is an important technique to acquire high quality cytogenetic images and facilitate the process of karyotyping so that the efficiency and accuracy of diagnosis can be improved. OBJECTIVE: This study investigated the impact of the condenser on cytogenetic image quality and system working performance using a prototype digital microscope image scanning system. METHODS: Both theoretical analysis and experimental validations through objectively evaluating a resolution test chart and subjectively observing large numbers of specimen were conducted. RESULTS: The results show that the optimal image quality and large depth of field (DOF) are …simultaneously obtained when the numerical aperture of condenser is set as 60%–70% of the corresponding objective. Under this condition, more analyzable chromosomes and diagnostic information are obtained. As a result, the system shows higher working stability and less restriction for the implementation of algorithms such as autofocusing especially when the system is designed to achieve high throughput continuous image scanning. CONCLUSIONS: Although the above quantitative results were obtained using a specific prototype system under the experimental conditions reported in this paper, the presented evaluation methodologies can provide valuable guidelines for optimizing operational parameters in cytogenetic imaging using the high throughput continuous scanning microscopes in clinical practice. Show more

Keywords: Optimizing operational parameters, microscope condenser, image sharpness, resolution, depth of field (DOF), cytogenetic imaging

DOI: 10.3233/ACP-130078

Citation: Analytical Cellular Pathology, vol. 36, no. 1-2, pp. 45-59, 2013

Article Type: Other

DOI: 10.3233/ACP-2013-0080

Citation: Analytical Cellular Pathology, vol. 36, no. 1-2, pp. 61-62, 2013