Automated image analysis of atomic force microscopy images of rotavirus particles

Venkataraman, Sankar; Allison, David P.; Qi, Hairong; Morrell‐Falvey, Jennifer L.; Kallewaard, N.L.; Crowe, James E.; Doktycz, Mitchel J.

doi:10.1016/j.ultramic.2006.01.014

Cited by 33 publications

(16 citation statements)

References 29 publications

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“…Out of many geometric features used by various authors in the literature 4,7 , it is observed that there are five geometric features, namely, circularity, compactness, eccentricity, tortuosity and length-width ratio, which provide better classification results. Hence, we have used these five features, which are defined as given below:…”

Section: Proposed Methodsmentioning

confidence: 99%

Automatic classification of bacterial cells in digital microscopic images

Hiremath

Bannigidad

2010

SPIE Proceedings

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The objective of the present study is to develop an automatic tool to identify and classify the bacterial cells in digital microscopic cell images. Geometric features are used to identify the different types of bacterial cells, namely, bacilli, cocci and spiral. The current methods rely on the subjective reading of profiles by a human expert based on the various manual staining methods. In this paper, we propose a method for bacterial classification by segmenting digital bacterial cell images and extracting geometric features for cell classification. The experimental results are compared with the manual results obtained by the microbiology expert and demonstrate the efficacy of the proposed method.

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Section: Proposed Methodsmentioning

confidence: 99%

Automatic classification of bacterial cells in digital microscopic images

Hiremath

Bannigidad

2010

SPIE Proceedings

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“…Moreover, high-quality AFM topography images of more than a dozen of pathological viruses were published to date [51,144,145,147]. Automated image analysis is very helpful in extracting the structural information, as was shown for rotaviruses [148]. The drawback of this approach may be the necessity of development of good surface adsorption methods working well for distinct pathogens [149].…”

Section: Afm Of Protein Complexes In Nanomedicine and Nanobio Techmentioning

confidence: 99%

AFM of biological complexes: What can we learn?

Gaczyńska

Osmulski

2008

Current Opinion in Colloid & Interface Science

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The term “biological complexes” broadly encompasses particles as diverse as multisubunit enzymes, viral capsids, transport cages, molecular nets, ribosomes, nucleosomes, biological membrane components and amyloids. The complexes represent a broad range of stability and composition. Atomic force microscopy offers a wealth of structural and functional data about such assemblies. For this review, we choose to comment on the significance of AFM to study various aspects of biology of selected nonmembrane protein assemblies. Such particles are large enough to reveal many structural details under the AFM probe. Importantly, the specific advantages of the method allow for gathering dynamic information about their formation, stability or allosteric structural changes critical for their function. Some of them have already found their way to nanomedical or nanotechnological applications. Here we present examples of studies where the AFM provided pioneering information about the biology of complexes, and examples of studies where the simplicity of the method is used toward the development of potential diagnostic applications.

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“…In addition, analytical methods like ELISA, Western blot, and transmission electron microscopy (TEM) are not able to clearly distinguish between DLP and TLP. Thus, novel techniques are required for the characterization of VLPs; multi-angle laser light scattering (MALLS) with liquid chromatography (Pattenden et al, 2005), analytical ultracentrifugation (Mach et al, 2006), field-flow fractionation (FFF) (Chuan et al, 2008), dynamic light scattering (DLS) (Pattenden et al, 2005), capillary electrophoresis (Mellado et al, 2008a), and atomic force microscopy (AFM) (Venkataraman et al, 2006) have been used for the characterization of RLPs or other VLPs. Mellado et al (2008a) developed a SDS-capillary gel electrophoresis (SDS-CGE) method for the analysis of purified DLP and TLP, which proved to be useful for apparent molecular weight estimation of each VP, degree of glycosylation in TLP and stoichiometric ratio determination.…”

Section: Introductionmentioning

confidence: 99%

Impact of physicochemical parameters on in vitro assembly and disassembly kinetics of recombinant triple‐layered rotavirus‐like particles

Mellado¹,

Mena²,

Lopes³

et al. 2009

Biotech & Bioengineering

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Virus-like particles constitute potentially relevant vaccine candidates. Nevertheless, their behavior in vitro and assembly process needs to be understood in order to improve their yield and quality. In this study we aimed at addressing these issues and for that purpose triple- and double-layered rotavirus-like particles (TLP 2/6/7 and DLP 2/6, respectively) size and zeta potential were measured using dynamic light scattering at different physicochemical conditions, namely pH, ionic strength, and temperature. Both TLP and DLP were stable within a pH range of 3-7 and at 5-25 degrees C. Aggregation occurred at 35-45 degrees C and their disassembly became evident at 65 degrees C. The isoelectric points of TLP and DLP were 3.0 and 3.8, respectively. In vitro kinetics of TLP disassembly was monitored. Ionic strength, temperature, and the chelating agent employed determined disassembly kinetics. Glycerol (10%) stabilized TLP by preventing its disassembly. Disassembled TLP was able to reassemble by dialysis at high calcium conditions. VP7 monomers were added to DLP in the presence of calcium to follow in vitro TLP assembly kinetics; its assembly rate being mostly affected by pH. Finally, DLP and TLP were found to coexist under certain conditions as determined from all reaction products analyzed by capillary electrophoresis. Overall, these results contribute to the design of new strategies for the improvement of TLP yield and quality by reducing the VP7 detachment from TLP.

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Automated image analysis of atomic force microscopy images of rotavirus particles

Cited by 33 publications

References 29 publications

Automatic classification of bacterial cells in digital microscopic images

Automatic classification of bacterial cells in digital microscopic images

AFM of biological complexes: What can we learn?

Impact of physicochemical parameters on in vitro assembly and disassembly kinetics of recombinant triple‐layered rotavirus‐like particles

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