• JSTS:Journal of Semiconductor Technology and Science
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• 제5권1호
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• pp.1-10
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• 2005

This paper presents the development of a microsystem for whole blood purification and electrophysiological analysis of the purified cells. Magnetophoresis using continuous diamagnetic capture (DMC) was utilized for whole cell purification and electrical impedance spectroscopy (EIS) was utilized for electrophysiological analysis of the purified cells. The system was developed on silicon and plastic substrates utilizing conventional microfabrication technologies and plastic microfabrication technologies. Using the magnetophoretic microseparator, white blood cells were purified from a sample of whole blood. The experimental results of the DMC microseparator show that 89.7% of the red blood cells (RBCs) and 72.7% of the white blood cells (WBCs) could be continuously separated out from a whole blood using an external magnetic flux of 0.2 T. EIS was used as a downstream whole cell analysis tool to study the electrophysiological characteristics of purified cells. In this work, primary cultured bovine chromaffin cells and human red blood cells were characterized using EIS. Further analysis capabilities of the EIS were demonstrated by successfully obtaining unique impedance signatures for chromaffin cells based on the whole cell ion channel activity.

• 대한기계학회논문집B
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• 제32권11호
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• pp.856-862
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• 2008

This paper presents the characterization of a continuous magnetophoretic microseparator for separating white and red blood cells from peripheral whole blood cells based on their native magnetic properties. The magnetophoretic microseparator separated the blood cells using a high gradient magnetic separation (HGMS) method without the use of additives such as magnetic beads or probing materials. Experimental results show that the paramagnetic capture mode microseparator can continuously separate out 93.5% of red blood cells and 97.4% of white blood cells from diluted whole blood, and the diamagnetic capture mode microseparator can continuously separate out 89.7% of red blood cells and 72.7 % of white blood cells by using applying an external magnetic flux of 0.2 T using a permanent magnet.

• Korea-Australia Rheology Journal
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• 제16권2호
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• pp.85-90
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• 2004

The present study investigated the deformability of red blood cells (RBC) and its effect on whole blood viscosity using a laser-diffraction slit-rheometer (LDSR). The LDSR has been recently developed with significant advances in laser-diffractometry design, operation and data analysis. While shear stress levels in a slit flow are continuously decreasing, both the deformation of red blood cells and the shear stress were simultaneously measured. Additionally, the viscosity of whole blood was measured using the LDSR. The present study found that the whole blood viscosity is strongly dependent on the RBC deformability. The less deformable the RBCs are, the higher the blood viscosity is.

• Asian-Australasian Journal of Animal Sciences
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• 제27권4호
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• pp.471-478
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• 2014

Investigating gene expression of immune cells of whole blood or peripheral blood mononuclear cells (PBMC) under polyinosinic:polycytidylic acid (poly I:C) stimulation is valuable for understanding the immune response of organism to RNA viruses. Quantitative real-time PCR (qRT-PCR) is a standard method for quantification of gene expression studies. However, the reliability of qRT-PCR data critically depends on proper selection of reference genes. In the study, using two different analysis programs, geNorm and NormFinder, we systematically evaluated the gene expression stability of six candidate reference genes (GAPDH, ACTB, B2M, RPL4, TBP, and PPIA) in samples of whole blood and PBMC with or without poly I:C stimulation. Generally, the six candidate genes performed a similar trend of expression stability in the samples of whole blood and PBMC, but more stably expressed in whole blood than in PBMC. geNorm ranked B2M and PPIA as the best combination for gene expression normalization, while according to NormFinder, TBP was ranked as the most stable reference gene, followed by B2M and PPIA. Comprehensively considering the results from the two programs, we recommended using the geometric mean of the three genes, TBP, PPIA and B2M, to normalize the gene expression of whole blood and PBMC with poly I:C stimulation. Our study is the first detailed survey of the gene expression stability in whole blood and PBMC with or without poly I:C stimulation and should be helpful for investigating the molecular mechanism involved in porcine whole blood and PBMC in response to poly I:C stimulation.

• Journal of Veterinary Science
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• 제24권1호
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• pp.11.1-11.14
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• 2023

Background: Peripheral blood mononuclear cells (PBMCs) are commonly used to assess in vitro immune responses. However, PBMC isolation is a time-consuming procedure, introduces technical variability, and requires a relatively large volume of blood. By contrast, whole blood assay (WBA) is faster, cheaper, maintains more physiological conditions, and requires less sample volume, laboratory training, and equipment. Objectives: Herein, this study aimed to develop a porcine WBA for in vitro evaluation of immune responses. Methods: Heparinized whole blood (WB) was diluted (non-diluted, 1/2, 1/8, and 1/16) in RPMI-1640 media, followed by phorbol myristate acetate and ionomycin. After 24 h, cells were stained for interferon (IFN)-γ secreting T-cells followed by flow cytometry, and the supernatant was analyzed for tumor necrosis factor (TNF)-α. In addition, diluted WB was stimulated by lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I:C), reference strain KCTC3557 (RS), field isolate (FI), of heat-killed (HK) Streptococcus suis, and porcine reproductive and respiratory syndrome virus (PRRSV). Results: The frequency of IFN-γ⁺CD3⁺ T-cells and concentration of TNF-α in the supernatant of WB increased with increasing dilution factor and were optimal at 1/8. WB TNF-α and interleukin (IL)-10 cytokine levels increased significantly following stimulation with LPS or poly I:C. Further, FI and RS induced IL-10 production in WB. Additionally, PRRSV strains increased the frequency of IFN-γ⁺ CD4^-CD8⁺ cells, and IFN-γ was non-significantly induced in the supernatant of re-stimulated samples. Conclusions: We propose that the WBA is a rapid, reliable, and simple method to evaluate immune responses and WB should be diluted to trigger immune cells.

• ETRI Journal
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• 제37권2호
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• pp.233-240
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• 2015

The novelty of this study resides in a 6"-wafer-level microfabrication protocol for a microdevice with a fluidic control system for the separation of circulating tumor cells (CTCs) from human whole blood cells. The microdevice utilizes a lateral magnetophoresis method based on immunomagnetic nanobeads with anti-epithelial cell adhesive molecule antibodies that selectively bind to epithelial cancer cells. The device consists of a top polydimethylsiloxane substrate for microfluidic control and a bottom substrate for lateral magnetophoretic force generation with embedded v-shaped soft magnetic microwires. The microdevice can isolate about 93% of the spiked cancer cells (MCF-7, a breast cancer cell line) at a flow rate of 40/100 mL/min with respect to a whole human blood/buffer solution. For all isolation, it takes only 10 min to process 400 mL of whole human blood. The fabrication method is sufficiently simple and easy, allowing the microdevice to be a mass-producible clinical tool for cancer diagnosis, prognosis, and personalized medicine.

• 한국자기학회지
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• 제24권1호
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• pp.28-33
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• 2014

최대 0.27 T 세기와 0.102 ms 펄스시간을 가지는 자기장 자극시스템을 이용하여 손에 인가한 후에 생혈액 분석을 통해 말초혈관에 흐르는 적혈구의 형태학적 변화를 조사하였다. 펄스자기장 자극 전 피실험자의 적혈구는 10여개 이상씩 서로 붙어있는 연전형성(連錢形成, rouleau formation)이 일어나서 적혈구의 움직임이 느려졌으나, 손에 10분간 자기장 인가한 후에 연전형성이 되었던 적혈구가 대부분 떨어져서 독립적인 움직임을 보이면서 운동이 활발해졌다. 또한 왼쪽 손에 펄스자기장을 10분 자극한 후에 자극받지 않은 오른쪽 손에서 채혈했을 때에도 연전현상이 똑같이 개선되었다. 이는 혈류흐름에 의해 체내에 모든 적혈구의 연전현상이 개선된 것으로 판단할 수 있다.

• 한국의료질향상학회지
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• 제3권2호
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• pp.70-85
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• 1997

Background : 6.1% of red blood cells and whole blood issued to the operating room was not transfused to the patients and discarded in Seoul National University Hospital in 1994. Objectives : We planned to set up an effective management program of blood in the operating room and we investigated whether this program could reduce the disposal rate of blood. Methods : We made a guideline of blood management in the operating room through a workshop. The guideline was revised after a preliminary application. The revised guideline was applied for 5 months from May to September in 1996. The disposal rate was compared before and after the installation of the new program. Results : 5,336 units of blood were issued to the operating room for 5 months. Disposal rate of red blood cells and whole blood was markedly reduced from 6.2% in May to 2.1% in September(p<0.05). The average disposal rate was 3.7% during the five months. Conclusion : We were able to reduce the disposal of unused blood in the operating room through the development and the application of a new blood management program.

• IMMUNE NETWORK
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• 제8권3호
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• pp.98-105
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• 2008

Background: Allergic inflammation was induced by activated Th2 lymphocytes, leading to IgE production and eosinophil activation. A Th2 disproportion was shown in atopic children soon after birth. During specific allergen stimulation, an increase of Th2 cells was observed in most cases. In this study, we prepared new screening "whole blood" system for searching the anti-atopic materials. Cytokine production and IgE secretion from whole blood system were assessed and we confirmed the results by using animal system. Methods: Pathological features in NC/Nga mice are similar to those observed in human atopic dermatitis. Whole blood from NC/Nga mouse was stimulated by using TNCB (Th2 activator) or candidate materials of anti-atopic dermatitis, and the production of cytokines (IL-4, IL-12, and IFN-${\gamma}$) were measured by ELISA. In order to confirm the results of whole blood system, in vivo test was done by using NC/Nga mice. Results: In whole blood system, LPS and extracts of green tea, hardy orange and onion induced the production of IL-12 and IFN-${\gamma}$ while they reduced the production of IL-4. Also, LPS and extracts of onion reduced IgE production. Though atopic dermatitis was observed from a mouse stimulated with TNCB, it was not when a mouse was co-stimulated in LPS or extracts of onion. The results are same as those observed in whole blood system. Conclusion: Whole blood system was simple and speedy methods for searching a materials compared with the conventional high-cost animal system. And the results using whole blood system was proved to be reliable in our experiments for screening anti-atopic material. We expect that the system can be applied to other experiments for searching similar materials.

• 한국방사선학회논문지
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• 제17권2호
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• pp.223-230
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• 2023

본 연구는 전신 방사선 조사 후 적혈구의 형태적 변화를 고찰하고자 하였다. 혈액 시료는 방사선 조사한 랫드(Rat)와 마우스의 적혈구를 사용하였다. 투과전자현미경 관찰결과, 5 Gy 조사 후 20일 지난 적혈구에서 대소부동증이 관찰되었다. 작은 적혈구에서는 삼각형, 테트라포드형의 형태가 관찰되었다. 일반적 크기의 적혈구에는 변형적혈구, 낫 모양의 겸상적혈구, 유극적혈구 등이 관찰되었다. 7 Gy 조사 후 20일 지난 적혈구는 분열적혈구가 관찰되었다. 주사전자현미경 관찰결과, 작은 적혈구들과 함께 누적적혈구가 관찰되었다. 또한 적혈구들이 서로 엉킨 상태의 군집된 형태도 확인되었다. 그 외 다각형의 모양, 반달 모양의 변형된 형태도 관찰되었다. 결론적으로 방사선 피폭에 의한 적혈구 연구는 수적 변화보다 변형된 형태의 병적 연구가 더 중요한 것으로 판단된다.