• Title/Summary/Keyword: pipette

Search Result 98, Processing Time 0.033 seconds

Roles of Nitric Oxide in Vestibular Compensation

  • Jeong, Han-Seong;Jun, Jae-Yeoul;Park, Jong-Seong
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.7 no.2
    • /
    • pp.73-77
    • /
    • 2003
  • The effects of nitric oxide on the vestibular function recovery following unilateral labyrinthectomy (UL) were studied. Sprague-Dawley male rats, treated with nitric oxide liberating agent sodium nitroprusside (SNP) and NOS inhibitor $N^G$-nitro-L-arginine methyl ester (L-NAME), were subjected to destruction of the unilateral vestibular apparatus, and then spontaneous nystagmus was observed in the rat. To explore the effects of nitric oxide on the neuronal excitability, whole cell patch clamp technique was applied on isolated medial vestibular nuclear neurons. The frequency of spontaneous nystagmus in SNP treated rats was lesser than that of spontaneous nystagmus in control animals. In contrast, pre-UL treatment with L-NAME resulted in a significant increase in spontaneous nystagmus frequency. In addition, SNP increased the frequency of spontaneous action potential in isolated medial vestibular nuclear neurons. Potassium currents of the vestibular nuclear neurons were inhibited by SNP. After blockade of calcium dependent potassium currents by high EGTA (11 mM) in a pipette solution, SNP did not inhibit outward potassium currents. 1H-[1,2,4] oxadiazolo [4,3-a] quinozalin-1-one (ODQ), a specific inhibitor of soluble guanylyl cyclase, inhibited the effects of SNP on the spontaneous firing and the potassium current. These results suggest that nitric oxide after unilateral labyrinthectomy would help to facilitate vestibular compensation by inhibiting calcium-dependent potassium currents through increasing intracellular cGMP, and consequently would increase excitability in ipsilateral vestibular nuclear neurons.

Culture Collection of Marine Microalgae (해양 미세조류의 배양수집)

  • Hur, Sung-Bum
    • Journal of Aquaculture
    • /
    • v.5 no.1
    • /
    • pp.81-91
    • /
    • 1992
  • The cultivation of marine microalgae as a live feed is essential for the seedling production of marine animals. It has some technical problems. Of these, the isolation and maintenance of the pure strains of microalgae from the nature are difficult for general mariculturists and researchers. To meet these problems, it needs to establish the culture collection of the microalgae in order to supply the strains to the demanders. In this research, 80 strains of microalgae were isolated from the coastal water of Korea by the methods of capillary pipette, plating on agar and dillution. A culture collection was established in the Department of Aquaculture, National Fisheries University of Pusan. The 117 strains of the microalgae maintained in the culture collection will be supplied to the demanders without any difficulties.

  • PDF

Cellular Force Sensing for Force Feedback-Based Biological Cell Injection (힘 피드백 기반의 세포조작을 위한 세포막 침습력 측정)

  • Kim, Deok-Ho;Yun, Seok;Kang, Hyun-Jae;Kim, Byung-Kyu
    • Transactions of the Korean Society of Mechanical Engineers A
    • /
    • v.27 no.12
    • /
    • pp.2079-2084
    • /
    • 2003
  • In biological cell manipulation, manual thrust or penetration of an injection pipette into an embryo cell is currently performed by a skilled operator, relying on visual feedback information only. Accurately measuring cellular forces is a requirement for minimally invasive cell injections. Moreover, the cellular force sensing is essential in investigating the biophysical properties for cell injury and membrane modeling studies. This paper presents cellular force measurements for the force feedback-based biomanipulation. Cellular force measurement system using piezoelectric polymer sensor is implemented to measure the penetration force of a zebrafish egg cell. First, measurement system setup and calibration are described. Second, the force feedback-based biomanipulation is experimentally carried out. Experimental results show that it successfully supplies real-time cellular force feedback to the operator at tens of uN and thus plays a main role in improving the reliability of biological cell injection tasks.

Cellular Force Measurement for Force Feedback-Based Biomanipulation (힘반향 기반의 바이오매니퓰레이션을 위한 세포 조작력 측정)

  • Kim, Duk-Ho;Kim, Byung-Kyu;Yoon, Seok;Kang, Hyun-Jae
    • Proceedings of the Korean Society of Precision Engineering Conference
    • /
    • 2003.06a
    • /
    • pp.237-240
    • /
    • 2003
  • In biological cell manipulation, manual thrust or penetration of an injection pipette into an embryo cell is currently performed by a skilled operator, relying on visual feedback information only. Accurately measuring cellular forces is a requirement for minimally invasive cell injections. Moreover, the cellular farce sensing is essential in investigating the biophysical properties for cell injury and membrane modeling studies. This paper presents cellular force measurements for the force feedback-based biomanipulation. Cellular force measurement system using piezoelectric polymer sensor is implemented to measure the penetration force of a zebrafish egg cell. First, measurement system setup and calibration are described. Second, the force feedback-based biomanipulation is experimentally carried out. Experimental results show that it successfully supplies real-time cellular force feedback to the operator at several tens of uN and thus plays a main role in improving the reliability of biological cell injection tasks.

  • PDF

Cryopreservation of Capsicum annum var. grossum using encapsulation/dehydration of apices produced in vitro

  • Senarath, Wtpsk;Lee, Kui-Jae;Rehman, S.;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
    • /
    • 2002.11b
    • /
    • pp.53-53
    • /
    • 2002
  • Shoot tips of in vitro propagated plantlets were cryopreserved using encapsulation/dehydration procedures. Shoot tips were excised under filter sterilized antioxidants solution (0.2M phosphate buffer, pH 5.7 supplemented with 5g/1 ascorbic acid and 15g/1 sodium borate). They were drawn up into a sterile 10 $\textrm{cm}^3$disposable pipette and were dropped into the culture medium with 2.5w/v Na-alginate, then into 100mM CaCl$_2$.2$H_2O$. Encapsulated shoot tips were transferred into 10㎤ of liquid culture medium with a range of sucrose concentrations (0.25-1.0M) and were incubated in dark for 24 hours in 18C at 40rpm. Beads were then dehydrated in silica gel for different time intervals (1-24 hours). Then they were freeze dried either rapidly (plunge directly into liquid N2 or in two stages (samples were kept at 20C for 10 minutes, then reduced to 35C at 1C per minute. Then, plunge into liquid $N_2$). The influence of sucrose and silica gel pre-treatment on pre- and post-freeze shoot growth was examined.(중략)

  • PDF

Studies on the Efficient Separation of Primordial Germ Cells and Introduction of Foreign DNA in the Chicken (닭에서 원시생식세포의 효율적 분리 및 외래 유전자 전이에 관한연구)

  • 정동기;한재용
    • Proceedings of the Korea Society of Poultry Science Conference
    • /
    • 1999.11a
    • /
    • pp.11-33
    • /
    • 1999
  • This study was conducted to determine the embryonic stages for the isolation of the highest number of PGCs and to improve PGCs enrichment method. The primordial germ cells(PGCs) from different sources of chick embryos were isolated. The embryonic stage having the highest number of PGCs from each sources was selected ; 1-day-old embryos for germinal crescent (stage 6-8), 2.5-day-old embryos for blood (stage 17-18) and 5.5-day-old embryos for gonad (stage 27-28). The number of PGCs from one embryonic germinal crescent, blood and gonad was about 87$\pm$1.8, 103$\pm$4.0, and 932$\pm$10.9, respectively. The viability of PGCs after Ficoll from each sources was similar, showing approximately 70%. the PGCs enrichment method was improved using Ficoll density gradient centrifugation. After this step the purity of PGCs from germinal crescent, blood, and gonad was 45$\pm$9.10%, 85$\pm$1.18%, and 86$\pm$0.19%, respectively. Also, PGCs were picked up by mouth pipette to improve the purity. This improved method for the separation of PGCs from different sources will serve as a useful too to preserve the foundation stocks of poultry and to produce germline chimeras.

  • PDF

Inhibition of the Desensitization of Canonical Transient Receptor Potential Channel 5 by Dimethyl Sulfoxide

  • Kim, Byung-Joo;So, In-Suk
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.11 no.5
    • /
    • pp.227-231
    • /
    • 2007
  • The classic type of transient receptor potential channel(TRPC) is a molecular candidate for $Ca^{2+}$-permeable cation channel in mammalian cells. TRPC5 is rapidly desensitized after activation by G protein-coupled receptor. Herein we report the effect of dimethyl sulfoxide(DMSO) on the desensitization of TRPC5. TRPC5 was initially activated by muscarinic stimulation with $50{\mu}M$ carbachol(CCh) and then decayed rapidly even in the presence of CCh(desensitization). DMSO in the pipette solution slowed the rate of this desensitization. Under the control conditions, TRPC5 current spontaneously declined to $6{\pm}1%$ of the initial peak amplitude 60 sec after CCh application and to $1{\pm}0.5%$ after 120 sec. But, in the presence of 0.01%, 0.1% and 1% DMSO, TRPC5 current spontaneously declined to $55{\pm}2%,\;68{\pm}1%\;and\;100{\pm}0.2%$ of the initial peak amplitude 60 sec after CCh application and to $38{\pm}2%,\;61{\pm}1%\;and\;100{\pm}1%$ after 120 see, respectively. The results suggest that DMSO can internally attenuate the desensitization of TRPC5 current through unknown mechanisms that remain to be elucidated.

Studies on Embryo Cryopreservation and Twinning by Embryo Transfer of Korean Native Cattle II. Transfer of Bisected Embryos and Production of Twin Calves (한우 수정란의 동결보존 및 쌍자생산에 관한 연구 II. 이분 수정란의 이식과 쌍자 생산)

  • 손동수;김일화;이동원;안병석;이광선;신형두;박노웅;최상용
    • Journal of Embryo Transfer
    • /
    • v.12 no.1
    • /
    • pp.91-102
    • /
    • 1997
  • This study was carried out to enhance the efficiency of Korean Native cattle embryos and establish the techniques for producing the twin calves. Bisected embryos without zona pellucida which were divided by simple method not using holding pipette or whole two embryos were transferred to recipients.The pedigrees of monozygotic twin calves produced by transfer of bisected pair embryos were identified. The results obtained were as follows ; The average successful bisection rate was 89.16%. The embryos of blastocyst stage (91.66%) were bisected successfully at significantly (P<0.05) higher rate, compared with the morula stage embryos (86.66%). The average survival rate of bisected embryos following 24 hours culture was 59.02%. The survival rate of morula stage embryos (62.50%) was significantly (P<0.05) higher than that of blastocyst stage embryos (55.5%). For the production of monozygotic twin calves, ten pairs of flesh or frozen demi-em- lymphocytes antigen, the twin calves produced by transfer of bisected pair embryos of Korean Native cattle were identified in pedigrees and confirmed as monozygotes.

  • PDF

Comparison of Developmental Efficiency of Murine Somatic Cell Nuclear Transfer Protocol

  • Moon, Jeonghyeon;Jung, Miran;Roh, Sangho
    • Journal of Embryo Transfer
    • /
    • v.32 no.3
    • /
    • pp.81-86
    • /
    • 2017
  • The Somatic cell nuclear transfer (SCNT) method can be applied to various fields such as species conservation, regenerative medicine, farming industries and drug production. However, the efficiency using SCNT is very low for many reasons. One of the troubles of SCNT is that it is highly dependent on the researcher's competence. For that reason, four somatic cell nuclear injection methods were compared to evaluate the effect of hole-sealing process and existence of cytochalasin B (CB) on efficiency of murine SCNT protocol. As a results, the microinjection with the hole-sealing process, the oocyte plasma membrane is inhaled with injection pipette, in HCZB with CB was presented to be the most efficient for the reconstructed in SCNT process. In addition, we demonstrated that the oocytes manipulated in Hepes-CZB medium (HCZB) with CB does not affect the developmental rate and the morphology of the blastocyst during the pre-implantation stage. For this reason, we suggest the microinjection involving hole-sealing in HCZB with CB could improve SCNT process efficiency.

Application of the modified handmade cloning technique to pigs

  • Lee, Eun Ji;Ji, Kuk Bin;Lee, Ji Hye;Oh, Hyun Ju;Kil, Tae Young;Kim, Min Kyu
    • Journal of Animal Science and Technology
    • /
    • v.63 no.2
    • /
    • pp.281-294
    • /
    • 2021
  • Although somatic cell nuclear transfer (SCNT) is frequently employed to produce cloned animals in laboratories, this technique is expensive and inefficient. Therefore, the handmade cloning (HMC) technique has been suggested to simplify and advance the cloning process, however, HMC wastes many oocytes and leads to mitochondrial heteroplasmy. To solve these problems, we propose a modified handmade cloning (mHMC) technique that uses simple laboratory equipment, i.e., a Pasteur pipette and an alcohol lamp, applying it to porcine embryo cloning. To validate the application of mHMC to pig cloning, embryos produced through SCNT and mHMC are compared using multiple methods, such as enucleation efficiency, oxidative stress, embryo developmental competence, and gene expression. The results show no significant differences between techniques except in the enucleation efficiency. The 8-cell and 16-cell embryo developmental competence and Oct4 expression levels exhibit significant differences. However, the blastocyst rate is not significantly different between mHMC and SCNT. This study verifies that cloned embryos derived from the two techniques exhibit similar generation and developmental competence. Thus, we suggest that mHMC could replace SCNT for simpler and cheaper porcine cloning.