• Journal of Life Science
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• v.17 no.2 s.82
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• pp.198-203
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• 2007

DNA transfection is a powerful tool for studying gene functions. The $Ca^{2+}$-phosphate precipitation remains one of the most popular and cost-effective transfection techniques. Mature neurons are more resistant to transfection than young ones and most other cell types, and easy to die if microenvironment changes. Here, we report a transfection protocol for mature neurons. The critical modifications are inclusion of glial cells in culture and careful control of $Ca^{2+}$-phosphate precipitation under microscope. Cerebral glial cells were grown until ${\sim}70-80%$ confluence in DMEM/10% horse serum, which was thereafter replaced with serum-free Neurobasal/Ara-C, and 319 hippocampal neurons were plated onto the glial layer Formation of fine $DNA/Ca^{2+}$-phosphate precipitates was induced using Clontech $CalPhos^{TM}$ Mammalian Transfection Kit, and the size ($0.5-1\;{\mu}m$ in diameter) and density(about 10 particles/$100\;{\mu}m^2$) were carefully controlled by the time of incubation in the medium. This modified protocol can be reliably applied for transfection of mature neurons that are maintained longer than two weeks in vitro, resulting in 10-15 healthy transfected neurons per a well of 24-well plates. The efficacy of the protocol was verified by punctate expression of $pEGFP-CaMKII{\alpha}$, a synaptic protein, and diffuse expression of pDsRed2. Our protocol provides a reliable method for transfection of mature neurons in vitro.

• Progress in Medical Physics
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• v.14 no.4
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• pp.211-217
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• 2003

The Principal component analysis (PCA) is a well-known data analysis method that is useful in linear feature extraction and data compression. The PCA is a linear transformation that applies an orthogonal rotation to the original data, so as to maximize the retained variance. PCA is a classical technique for obtaining an optimal overall mapping of linearly dependent patterns of correlation between variables (e.g. neurons). PCA provides, in the mean-squared error sense, an optimal linear mapping of the signals which are spread across a group of variables. These signals are concentrated into the first few components, while the noise, i.e. variance which is uncorrelated across variables, is sequestered in the remaining components. PCA has been used extensively to resolve temporal patterns in neurophysiological recordings. Because the retinal signal is stochastic process, PCA can be used to identify the retinal spikes. With excised rabbit eye, retina was isolated. A piece of retina was attached with the ganglion cell side to the surface of the microelectrode array (MEA). The MEA consisted of glass plate with 60 substrate integrated and insulated golden connection lanes terminating in an 8${\times}$8 array (spacing 200 $\mu$m, electrode diameter 30 $\mu$m) in the center of the plate. The MEA 60 system was used for the recording of retinal ganglion cell activity. The action potentials of each channel were sorted by off­line analysis tool. Spikes were detected with a threshold criterion and sorted according to their principal component composition. The first (PC1) and second principal component values (PC2) were calculated using all the waveforms of the each channel and all n time points in the waveform, where several clusters could be separated clearly in two dimension. We verified that PCA-based waveform detection was effective as an initial approach for spike sorting method.

• Journal of Intelligence and Information Systems
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• v.22 no.2
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• pp.57-79
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• 2016

It is a rising trend that the number of users using one of the social media channels, the Social Network Service, so called the SNS, is getting increased. As per to this social trend, more companies have interest in this networking platform and start to invest their funds in it. It has received much attention as a tool spreading and expanding the message that a company wants to deliver to its customers and has been recognized as an important channel in terms of the relationship marketing with them. The environment of media that is radically changing these days makes possible for companies to approach their customers in various ways. Particularly, the social network service, which has been developed rapidly, provides the environment that customers can freely talk about products. For companies, it also works as a channel that gives customized information to customers. To succeed in the online environment, companies need to not only build the relationship between companies and customers but focus on the relationship between customers as well. In response to the online environment with the continuous development of technology, companies have tirelessly made the novel marketing strategy. Especially, as the one-to-one marketing to customers become available, it is more important for companies to maintain the relationship marketing with their customers. Among many SNS, Facebook, which many companies use as a communication channel, provides a fan page service for each company that supports its business. Facebook fan page is the platform that the event, information and announcement can be shared with customers using texts, videos, and pictures. Companies open their own fan pages in order to inform their companies and businesses. Such page functions as the websites of companies and has a characteristic of their brand communities such as blogs as well. As Facebook has become the major communication medium with customers, companies recognize its importance as the effective marketing channel, but they still need to investigate their business performances by using Facebook. Although there are infinite potentials in Facebook fan page that even has a function as a community between users, which other platforms do not, it is incomplete to regard companies' Facebook fan pages as communities and analyze them. In this study, it explores the relationship among customers through the network of the Facebook fan page users. The previous studies on a company's Facebook fan page were focused on finding out the effective operational direction by analyzing the use state of the company. However, in this study, it draws out the structural variable of the network, which customer committment can be measured by applying the social network analysis methodology and investigates the influence of the structural characteristics of network on the business performance of companies in an empirical way. Through each company's Facebook fan page, the network of users who engaged in the communication with each company is exploited and it is the one-mode undirected binary network that respectively regards users and the relationship of them in terms of their marketing activities as the node and link. In this network, it draws out the structural variable of network that can explain the customer commitment, who pressed "like," made comments and shared the Facebook marketing message, of each company by calculating density, global clustering coefficient, mean geodesic distance, diameter. By exploiting companies' historical performance such as net income and Tobin's Q indicator as the result variables, this study investigates influence on companies' business performances. For this purpose, it collects the network data on the subjects of 54 companies among KOSPI-listed companies, which have posted more than 100 articles on their Facebook fan pages during the data collection period. Then it draws out the network indicator of each company. The indicator related to companies' performances is calculated, based on the posted value on DART website of the Financial Supervisory Service. From the academic perspective, this study suggests a new approach through the social network analysis methodology to researchers who attempt to study the business-purpose utilization of the social media channel. From the practical perspective, this study proposes the more substantive marketing performance measurements to companies performing marketing activities through the social media and it is expected that it will bring a foundation of establishing smart business strategies by using the network indicators.