• Title/Summary/Keyword: Micropipette

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Electrochemical Detection of Single Nucleotide Polymorphism (SNP) Using Microelectrode Array on a DNA Chip (미소전극어레이형 DNA칩을 이용한 유전자다형의 전기화학적 검출)

  • 최용성;권영수;박대희
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.53 no.5
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    • pp.286-292
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    • 2004
  • In this study, an integrated microelectrode array was fabricated on glass slide using microfabrication technology. Probe DNAs consisting of mercaptohexyl moiety at their 5-end were spotted on the gold electrode using micropipette or DNA arrayer utilizing the affinity between gold and sulfur. Cyclic voltammetry in 5mM ferricyanide/ferrocyanide solution at 100 ㎷/s confirmed the immobilization of probe DNA on the gold electrodes. When several DNAs were detected electrochemically, there was a difference between target DNA and control DNA in the anodic peak current values. It was derived from specific binding of Hoechst 33258 to the double stranded DNA due to hybridization of target DNA. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic System.

A Study on Electrical Properties of Dendrimer (미소전극형 DNA칩 어레이를 이용한 유전자의 검출)

  • Choi, Yong-Sung;Lee, Kyung-Sup
    • Proceedings of the KIEE Conference
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    • 2006.07c
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    • pp.1324-1326
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    • 2006
  • In this study, an integrated microelectrode array was fabricated on glass slide using microfabrication technology. Probe DNAs consisting of mercaptohexyl moiety at their 5-end were spotted on the gold electrode using micropipette or DNA arrayer utilizing the affinity between gold and sulfur. Cyclic voltammetry in 5mM ferricyanide/ferrocyanide solution at 100 mV/s confirmed the immobilization of probe DNA on the gold electrodes. When several DNAs were detected electrochemically, there was a difference between target DNA and control DNA in the anodic peak current values. It was derived from specific binding of Hoechst 33258 to the double stranded DNA due to hybridization of target DNA. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic system.

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Characteristic Analysis of Electrowetting on Dielectric Layer (절연층에 따른 액적의 전기습윤 특성 분석)

  • Choi, Jin Ho;Kim, Gyu man
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.18 no.8
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    • pp.38-43
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    • 2019
  • Electrowetting on dielectric (EWOD) is a unique method of shape control of small-volume droplets in microfluidic biochips that relies on modification of surface wetting characteristics using electrical methods. In this study, the droplet shape control on various dielectric surfaces by the EWOD and the effect of droplets on the contact angle as well as the shape were investigated. The droplet used in the experiment was on a sample substrate with $5{\mu}l$ of de-ionized water (DIW) using a micropipette, and wettability was measured with a contact angle meter. This study is expected to be helpful for the development of various micro-total-analysis-systems (${\mu}TAS$) and microfluidic systems with MEMS technology.

Improvement of Antigen Blotting in a Tissue Blot Immunobinding Assay for the Detection of Two Chili Pepper Viruses

  • Han, Jung-Heon;Shin, Jun-Sung;Kim, Young-Ho;Kim, Byung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1885-1889
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    • 2007
  • The tissue blot immunobinding assay (TBIA) is widely used for the detection and localization of plant viruses in various plant tissues. The basic experimental procedures of TBIA sampling and blotting were simplified using commercially available micropipette tips. This method was termed the ring-blot immunobinding assay (R-BIA), as the blot on the membrane forms a ring shape. The detection efficacy of R-BIA was tested for two chili pepper viruses, pepper mild mottle tobamovirus (PMMoV) and pepper mottle potyvirus (PepMoV), following the optimized serological procedures of TBIA (length of the incubation period and BSA concentration, and primary and secondary antibodies). Sensitivity of the R-BIA was about 1 ng/ml of purified PMMoV in pepper leaf sap from a healthy pepper plant. R-BIA also showed high specificity in the detection of PMMoV and PepMoV. Moreover, the modified sampling and blotting procedures were simpler and more reliable than other TBIA methods (such as whole-leaf blotting and crushed-leaf blotting), suggesting that the R-BIA may be used for medium- to large-scale detection of plant viruses in laboratories with minimal facilities.

Modification of Oropharyngeal Aspiration Technique for Mouse Using Syringe Pump

  • Kim, Jin-Sung;Yang, Mi-Jin;Han, Sung-Gu;Kim, Choong-Yong;Han, Sang-Sup;Song, Chang-Woo
    • Toxicological Research
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    • v.23 no.3
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    • pp.239-244
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    • 2007
  • Respirable particles cause many occupational and environmental diseases of the lung. To study these diseases, laboratory animals are often exposed to these particles. Inhalation and instillation are the well-known techniques for experimental exposures of the lung to respirable particles. Recently, another technique called oropharyngeal aspiration (OPA) has been introduced for exposing the lung to pathogens and/or particles. The conventional OPA technique for the mouse is generally carried out using a micropipette with a fixed slant board. In order to modify the conventional OPA in this study, anesthetized mice were placed on an adjustable slant board, a syringe pump was used to deliver the solution to the oropharynx, and the mice were allowed to recover in vertically positioned tubes for 6 minutes until fully awaked. Most importantly, the whole process of OPA could be carried out simply by an examiner. This modified OPA technique was validated by exposing the mouse lung to Evans Blue dye with a success rate of 95%.

PDMS/Glass Serpentine Microchannel Chip for PCR with Bubble Suppression in Sample Injection (시료주입시 기포발생이 억제된 반응조 형태의 중합효소연쇄반응용 PDMS/유리 바이오칩)

  • Cho Chul-Ho;Cho Woong;Hwang Seung-Yong;Ahn Yoo-Min
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.30 no.10 s.253
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    • pp.1261-1268
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    • 2006
  • This paper reports low-cost microreactor $(10{\mu}{\ell})$ biochip for the DNA PCR (polymerase chain reaction). The microbiochip $(20mm{\times}28mm)$ is a hybrid type which is composed of PDMS (polydimethylsiloxane) layer with serpentine micochannel $(360{\mu}m{\times}100{\mu}m)$ chamber and glass substrate integrated with microheater and thermal microsensor. Undesirable bubble is usually created during sample loading to PMDS-based microchip because of hydrophobic chip surface. Created bubbles interrupt stable biochemical reaction. We designed improved microreactor chamber using microfluidic simulation. The designed reactor has a coner-rounded serpentine channel architecture, which enables stable injection into hydrophobic surface using micropipette only. Reactor temperature needed to PCR reaction is controlled within ${\pm}0.5^{\circ}C$ by PID controller of LabVIEW software. It is experimentally confirmed that SRY gene PCR by the fabricated microreactor chip is performed for less than 54 min.

CLINICAL STUDY OF DESENSITIZER FOR TREATMENT OF DENTINAL HYPERSENSITIVITY (상아질 지각과민 처치제의 임상적 효과에 대한 연구)

  • Cho, Kyung-Mo;Shin, Dong-Hoon
    • Restorative Dentistry and Endodontics
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    • v.25 no.2
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    • pp.219-224
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    • 2000
  • The purpose of this study is for evaluating the effect of MS Coat desensitizing agent in clinical situation. In this study total 60 teeth of 30 patients who is feeling hypersensitivity after periodontal surgery were treated with MS Coat desensitizing agent, and 20 teeth with saline solution for placebo effects and evaluated. All teeth were cleaned with rubber cup and pumice, after then the antibacterial agent was applied for 20 seconds and simply isolated using cotton roll Desensitizing agent was applied with a specific instrument in the manufacturers package by abrading motion for 10 seconds and re-done 8 times. Tactile stimulus with sharp explorer, air stimulus with syringe of dental unit, $7^{\circ}C$ cold water stimulus using micropipette applicator was done to evaluate hypersensitivity before apply test agent, immediate after application, 1 week after, and 3 month after application. The degree of hypersensitivity was recorded using 100mm Visual Analogue Scale and compared between group and evaluation times. From this clinical study sensitivity to the stimulus was significantly induced in both groups and MS Coat made a significant reduction in VAS score than placebo group did(p<.01). The results of this study could demonstrate that MS Coat desensitizing agent can use effectively to heat dentinal hypersensitivity.

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Studies on the use of computer aided semen analysis(CASA) technology for fertility prediction in Korean native cattle (한우에서 Computer aided semen analysis(CASA) 기법을 이용한 수태율 예측에 관한 연구)

  • Lee, Kang-nam;Lee, Byeong-chun;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.882-897
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    • 1998
  • This experiment was conducted to predict the effects of motional characteristics on the fertility of Korean native cattle(KNC) by using CASA technology and in vitro fertilization system. Twenty-six KNC frozen semen straws were obtained from Korean KNC improvement department, livestock improvement main division, national livestock cooperatives federation in Korea. Specimens were allowed to thaw at $37^{\circ}C$ for 30 sec in water bath. Semen analysis was performed on semen image analysis system(SIAS, Medical supply, Korea) adjusted to the gate settings and used the semen droplet ($5{\mu}l$) placed on Makler counting chamber(Sefi medical instrument, Israel) prewarmed at $37^{\circ}C$. The same person used the same micropipette to fill the Makler counting chamber. A total of 150 or more of sperms were analysed in each specimen by a single trained person by scanning at least 5 to 10 fields. The oocytes collection, in vitro maturation, IVF, in vitro culture and determination of the cleavage rate were performed by the technique, as described by Hwang et al (1997). Statistical analysis was done by linear regression with use of the Sigma plot program on a IBM personal computer. The cleavage rate in vitro fertilized oocyte was significantly correlated(P<0.05) with MOT, VCL, VSL, VAP, ALH, BCF and MAD, but not CON, LIN, STR, WOB, DNM, DNC and HYP in regressional analysis. The results show that some kinematic characteristics of frozen-thawed semen by CASA can be predict the fertility in in vitro model system.

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Real-Time Force Sensing in the Envelope of Zebrafish Egg during Micropipette Penetration

  • Yun, Seok;Kim, Deok-Ho;Kim, Byung-Kyu;Lee, Sang-Ho;Park, Gwi-Tae
    • 제어로봇시스템학회:학술대회논문집
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    • 2003.10a
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    • pp.2451-2456
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    • 2003
  • In biological cell manipulation, manual thrust or penetration of an injection pipette into an egg is currently performed by a skilled operator, relying only on visual feedback information. Massive load of various micro injection of either genes, fluid or cells in the postgenomic era calls a more reliable and automatic micro injection system that can test hundreds of genes or cell types at a single experiment. We initiated to study cellular force sensing in zebrafish eggs as the first step for the development of a more controllable micro injection system by any inexperienced operator. Zebrafish eggs at different developmental stages were collected and an integrated biomanipulation system was employed to measure cellular force during penetrating the egg envelope, the chorion. First of all, the biomanipulation system integrated with cellular force sensing instrument is implemented to measure the penetration force of cell membranes and characterize mechanical properties of zebrafish embryo cells. Furthermore, implementation of cellular force sensing system and calibration are presented. Finally, the cellular force sensing of penetrating cell membranes at each developmental stages was experimentally performed. The results demonstrated that the biomanipulation system with force sensing capability can measure cellular force at real-time while the injection operation is undergoing. The magnitude of the measured force was in the range of several hundreds of uN. The precise real-time measurement should provide the first step forwards for the development of an automatic and reliable injection system of various materials into biological cells.

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Development of a New Improvement and Multiplication System in Domestic Animals Using a Embryonic Manipulation Technique I. Effect of Maturation Time on the Extrusion Rate of First Polar Body and the Enucleation Rate of Bovine Follicular Oocytes (세포조작 기술을 이용한 새로운 축산개량증식 체계 개발 I. 소 난포란의 성숙시기가 제 1극체 출현율과 핵제거율에 미치는 영향)

  • 임경순;김현종;오성종;양보석
    • Korean Journal of Animal Reproduction
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    • v.19 no.3
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    • pp.181-189
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    • 1995
  • In this study, methods on fabrication of microtool and setting of micromanipulator were examined and relationship between first polar body extrusion rate and maturation time of follicular oocyte, enulceation rae and repetition of trial, and enucleation rate and maturation period were investigated. The results are as follows: 1. Suitable outside diameter of micropipette tube was 1mm. Holding pipette with less than diameter of oocyte was fitred for manipulation, and zona dissection needle was easily operated when its sharp-point had diameter of about 8 ${\mu}{\textrm}{m}$ and length of 300${\mu}{\textrm}{m}$. The injection pipette with 20~35${\mu}{\textrm}{m}$ outside diameter was adequate for injection of blastomere into perivitelline space. 2. Separation of blastomere was effective when zona pellucida had cut with zonadissection needle and the embryo was pipetted gently with the pipette that had narrower diameter than that of embryo until separation of blastomeres had completed. 3. The extrusion rate of first polar body was 78% during 20~24% hours incubation for maturation. 4. According to repetitions of micromanipulation, the enucleation rate was increased to 85% and the time required for enucleation of a oocyte was shortened to 3 min. 5. The extrusion rate of first polar body and enucleation rate were 82 and 76% respectively, in the group of the oocytes cultured for 22 hours. However in the group cultured for 24 hours, the extrusion rate of first polar body and enucleation rate were 53 and 100% respectively.

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