저자는 아낌없이 H1 MSCS, 박사 팀의 해커 박사 Gouqing 노래와 위스콘신 순환기 생리학 대학의 양 질 코흐를 제공하는 박사 Peiman Hemmati을 (의학 계열, 위스콘신 - 매디슨 대학) 감사하고 싶습니다 마우스 수술을 수행하기위한 핵심 시설. 이 작품은 브라이언 프리맨과 NIH R21 HL089679에 대학원 연구 활동을 통해 국립 과학 재단 (National Science Foundation)에 의해 지원되었다.

1. 공여 세포 Transfection <ol><li> 수확 mesenchymal 줄기 세포 (친절하게 박사 Peiman Hematti 기증한 H1 배아의 줄기 세포에서 파생된 MSCS; 또는 생체내의 퓨즈로 가상 어떤 종류의 셀 타입은 고용 수) 70 - 1X 트립신과 합류 80 % (Mediatech, 5 분 매나 사스 VA는.) α - 가상 완료 매체 (항생제 무료, Invitrogen, Carlsbad CA) 18 트립신을 Inactivate. 5 분 300 XG에 원심 분리기. </li><li> 조심스럽게 뜨는 대기음하고 다시 중지 펠렛 1X PBS 1 ML에서 hemacytometer를 사용하여 셀을 계산합니다. </li><li> 1.5 ML의 원심 관에 전송 1.5 X 10 6 세포. 5 분 300 XG에 원심 분리기. </li><li> 조심스럽게 뜨는 기음. R 버퍼 (네온 Transfection 시스템, Invitrogen)와 6 μg (2 μg / 5.0 × 10 5 세포) 300 μL에 Resuspend 펠렛 p231 pCMVe - betaAc - STOP 룩 (Addgene, 캠브리지, MA). E 버퍼의 장소 3 ML (Invitr제조 업체의 절차에 따라 electroporation 도킹 포트 (네온 Transfection 시스템, Invitrogen)에 ogen). </li><li> 100 μL 네온 pipet 팁에 휴대 플라스미드 솔루션을 전송 20 MS와 1,500 볼트의 크기의 펄스 기간과 electroporate. 장소는 9.7 ML α - 전체 가상 매체를 포함 15 ML 원뿔 튜브에 세포를 electroporated. </li><li> 10 ML의 총 볼륨을 얻을 1.5 두 가지 추가 시간 및 수영장 transfected 세포 단계를 반복합니다. 10 ML α - 전체 가상 매체를 포함하는 T175 플라스크에 10 ML의 세포 현탁액을 (1.5 X 10 6 셀) 추가합니다. electroporation 후 세포 생존 능력은 T175 약 4.5 × 10 5 가능한 세포를 양보하기 위해, 약 30 %입니다. </li><li> 변경 transfection 다음 중간 24시간 α - 가상 완료되었습니다. </li><li> 80 % 합류 (~ 2 - - 삼일 electroporation 후) 수확 70 세포를 transfected. hemacytometer를 사용하여 셀 카운트를 수행하고 1.0 × 10 6 농도에서 세포를 resuspendα - 가상 완료 매체의 cells/50 μL. 최소화 시간 세포 주입하기 전에 세포 사망을 줄이기 위해 서스펜션에 보냅니다. </li></ol> 2. Intramyocardial 사출 <ol><li> 모든 셀에 constitutively 표현 Cre recombinase (B6.C - TG (CMV - cre) 1Cgn / J, 잭슨 연구소, 바 하버 수 있도록 설계 유전자 변형 생쥐에 (피닉스 제약 주식 회사, 세인트 조셉, MO) isoflurane으로 마취를 유도 ME). </li><li> 헤어 가위 또는 화학 헤어 리무버를 사용하여 가슴 지역에서 머리를 제거합니다. </li><li> 150 μL의 뇌졸중 볼륨 분당 130 심호흡 - 120에서 마우스 환풍기에 18 게이지 카테터 (Becton 디킨슨 &amp; CO, 프랭클린 호수 NJ)와 장소 Intubate. </li><li> 따라서 thoracotomy을 생산 번째 늑간 공간에 걸쳐 가로 절개를합니다. </li><li> 심장 떠올리, 1 ML 주사기 (Temuro 의료 법인, 서머셋, 뉴저지) 및 28 게이지 바늘 (Bect를 사용하여 transfected 세포 현탁액의 두 25 μL 주사를에 디킨스 &amp; CO, 프랭클린 호수 NJ). intramyocardial 주사를 완화하고 장기에 과도한 손상을 방지하기 위해 바늘 머리 ~ 90도 구​​부. </li><li> 주사 후, 갈비뼈 근육 레이어를 닫습니다 흡수성 봉합 (예 : vicryl)을 사용합니다. 봉합사의 피부는 4-0 나일론 또는 실크를 사용하여 마감했다. </li><li> 마취 및 extubate에서 회복하기 위해 마우스를 허용합니다. </li><li> 제어 그룹은, Cre 마우스가 untransfected 세포와 transfected 세포를 (또는, Cre 마우스는 퓨즈에 쉽게 무너지는 사람이 아니다 transfected 세포를 수신) 수신 야생 타입 마우스의 동일한 농도를받는 중간 주사를 받고 Cre 마우스를 포함해야합니다. </li></ol> 3. Biophotonic 이미징 <ol><li> 이미징, intraperitoneally은 (IP) 15 밀리그램 / ML D - Luciferin (캘리퍼스 생명 과학, Hopkinton, MA)의 마우스 체중 g 당 10 μL를 주입 다섯 데 15 분 전. </li><li> 유도 4 %로 isoflurane을 통해 생쥐에 마취를 유도D 1 - 유지 보수 2 %. </li><li> 안면이 1 관리와 이미징 상자에 나태한 장소 마우스 - 유지 보수 마취 (Xenogen Biophotonic 이미징 시스템, Hopkinton, MA)에 isoflurane 2퍼센트. 여러 생쥐는 동시에 몇 군데하실 수 있습니다. 발광 신호를 쉽게 비교 실험 쥐 이미지 가짜 제어 마우스. </li><li> 생활 이미지 소프트웨어 (Xenogen)를 사용하여 적절한 노출 시간 (일반적으로 60 초, 결과를 참고)으로 설정합니다. 쥐를 적합하고 감도의 변화를 방지하기 위해 영상을 통해 지역 일관성 유지하기 이미지의 영역을 설정합니다. 4.5 cm에서 제목 높이를 설정합니다. </li><li> 보기 필드 내에서 마우스 또는 마우스에 대응하는 발광 강도 신호를 수집하고 수정되지 않은 이미지 파일을 저장합니다. 프로세스 이미지 컨트롤 또는 unmanipulated 마우스에 해당하는 배경 신호를 제거할 수 있습니다. 배경 위의 강도 값은 동물 내에서 융합 세포에 해당합니다. 강도 분석은 생활 이미지 소프트웨어 (Xenogen) 또는 오픈 소스 이미지 분석 소프트웨어를 사용하여 진행하실 수 있습니다. 티ypically, 관심 지역은 (ROI) 동물과 실험 사이의 강도 데이터를 비교하기 위해 선택됩니다. </li></ol> 4. 대표 결과 Xenogen Biophotonic 이미징 시스템의 감도, constitutively C57/Bl6 마우스 (잭슨 연구소)의 myocardium에게 전달되었다 (231 룩 - D3H1, Xenogen) 루시페라제 표현하는 세포 라인을 확인하려면 다음과 같이하십시오. 셀은 1 X의 농도에 주입했다 10 6, 1 × 10 3, 또는 1 X 10 1 세포. 여섯 시간 세포 전달 후, 마우스가 luciferin와 intraperitoneally을 주입하고 Xenogen 시스템을 사용하여 몇 군데 있었다. 구체적인 신호가 1,000 세포 (몇 군데 2 6의 생쥐, 그림 2)로 감지하지만, 검색은 10,000 셀 (6 군데 생쥐 6)와 함께보다 안정되었습니다 수 있습니다. 중요한 것은,이 연구는 또한 루시페라제 - 표현 세포 및 신호 강도의 개수 사이에 거친 상관 관계를 수립 하였다. Cre - 표현 생쥐의 myocardium에 전달했다. 셀 배달 후 약 일주일이 생쥐 먼저 D - luciferin 주입없이 Xenogen 시스템을 사용하여 몇 군데 있었다. 으로 효소 기판없이 예상, 더 강도 신호는 (그림 3) 발견되지 않았습니다. 다음, D - luciferin는 intraperitoneally 및 루시페라제 강도에 해당하는 신호를 주입했고 따라서 세포 융합은 테스트를 두 넷의 생쥐에서 발견되었다. 비슷한 신호가이 경우에는 연구가 중심과 주변 조직의 평가 수 있도록 종료되었습니다. MSC - 결합 융합 제품은 생체내 유지 수있는 제안, 나중에 일주 (그림 3) 발견했지만, 하나는 구상 수있는 장기 분석 및 유지 보수, 확산을 추적하기 위해 더 자주 이미징마우스의 융합 제품 개발 아마 마이 그 레이션. <img alt="그림 1" src="/files/ftp_upload/3581/3581fig1.jpg" /> 그림 1. VIVO에서 세포 융합을 감지하는 기술의 개략도. 간의 융합이 Cre가 - 표현 마우스 세포 및 발생하는 floxed 루시페라제 플라스미드를 표현 이식 세포를 루시페라제가 표현되는 경우. 루시페라제는 Xenogen Biophotonic 이미징 시스템 (19에서 적응)를 사용하여 마우스를 마우스로 효소 기판, D - luciferin를 주입 후 영상에 의해 감지 수 있습니다 <img alt="그림 2" src="/files/ftp_upload/3581/3581fig2.jpg" /> 그림 2. 의 감지 감도 루시페라제 - 표현 biophotonic 영상과 심장 조직에서 세포를. constitutively (231 룩 - D3H1, Xenogen) 루시페라제 표현 세포 라인은 다양한 전체 휴대폰 번호에서 C57/Bl6 생쥐의 intramyocardial 공간으로 전달했다. 생쥐의 대표 이미지는 다시1 × 10 6, 1 X 10 3 1 X 10 1 세포 (오른쪽 왼쪽) ceiving하면 표시됩니다, 이미지는 대략 6 시간 후에 주사를 실시했다. <img alt="그림 3" src="/files/ftp_upload/3581/3581fig3.jpg" /> 그림 3. 세포 융합되었음을 암시 VIVO의 발광에의 부량. MSCS는 플라스미드 LoxP - 그만 - LoxP - 루시페라제과 Cre - 표현 생쥐의 myocardium 배달과 transfected되었다. 약 일주일에 2 주 세포 전달 후, Cre 마우스는 세포 융합을 나타내는 발광의 강도를 측정하는 Xenogen Biophotonic 이미징 시스템을 사용하여 몇 군데 있었다. 17일 세포 전달 후 사기와 생쥐 1-4 발광의 사진과 강도의 (A) 중첩 (오른쪽 왼쪽). (B) 사기와 생쥐 1-4 발광의 강도 (오른쪽 왼쪽) 휴대폰 전송 후 17일. 관심 지역은 사출 사이트 및 강도 리터에 해당하는 (노란색)을 선택했습니다evels는 ImageJ (무료 소스) 소프트웨어를 사용하여 20으로 결정됩니다. (C) 발광의 강도는 모든 실험 조건에 대한 사기 마우스의 관심 동일한 지역에 정상화되었다. 일주에서 마우스 3과 4는 마우스 세포와 이식 MSC의 자발적인 융합을 제안 긍정적인 발광 신호를 보여주었다. 신호는 2 주에서 마우스를 지속 3. 마우스 3에 해당하는 신호의 기관 고유의 로컬 리 제이션을 확인하려면, 흉강는 노출 및 기본 기관 excised하고 몇 군데했습니다. 사진 및 마우스 3 발광의 강도 (D) 중첩. 소장의 강도 신호의 참고 지방화. 마우스 3 발광의 (E) 강도. 사진과 가짜 마우스 발광의 강도 (F) 중첩. 가짜 마우스의 발광의 (G) 강도.

<ol>
	<li>Bernirschke, K. K. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Pathology of the Human Placenta. , (2000).</li><li>Hoshina, M., Boothby, M., Boime, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytological+localization+of+chorionic+gonadotropin+alpha+and+placental+lactogen+mRNAs+during+development+of+the+human+placenta.">Cytological localization of chorionic gonadotropin alpha and placental lactogen mRNAs during development of the human placenta.</a> J. Cell. Biol. 93, 190-198 (1982).</li><li>Johansen, M., Redman, C. W., Wilkins, T., Sargent, I. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trophoblast+deportation+in+human+pregnancy--its+relevance+for+pre-eclampsia.">Trophoblast deportation in human pregnancy--its relevance for pre-eclampsia.</a> Placenta. 20, 531-539 (1999).</li><li>Redman, C. W., Sargent, I. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Placental+debris,+oxidative+stress+and+pre-eclampsia.">Placental debris, oxidative stress and pre-eclampsia.</a> Placenta. 21, 597-602 (2000).</li><li>Ogle, B. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spontaneous+fusion+of+cells+between+species+yields+transdifferentiation+and+retroviral+transfer+in+vivo.">Spontaneous fusion of cells between species yields transdifferentiation and retroviral transfer in vivo.</a> FASEB. J. 18, 548-550 (2004).</li><li>Nygren, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone+marrow-derived+hematopoietic+cells+generate+cardiomyocytes+at+a+low+frequency+through+cell+fusion,+but+not+transdifferentiation.">Bone marrow-derived hematopoietic cells generate cardiomyocytes at a low frequency through cell fusion, but not transdifferentiation.</a> Nat. Med. 10, 494-501 (2004).</li><li>Nygren, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myeloid+and+lymphoid+contribution+to+non-haematopoietic+lineages+through+irradiation-induced+heterotypic+cell+fusion.">Myeloid and lymphoid contribution to non-haematopoietic lineages through irradiation-induced heterotypic cell fusion.</a> Nat. Cell. Biol. 10, 584-592 (2008).</li><li>Kutschka, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adenoviral+human+BCL-2+transgene+expression+attenuates+early+donor+cell+death+after+cardiomyoblast+transplantation+into+ischemic+rat+hearts.">Adenoviral human BCL-2 transgene expression attenuates early donor cell death after cardiomyoblast transplantation into ischemic rat hearts.</a> Circulation. 114, I174-I180 (2006).</li><li>Min, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+bioluminescence+imaging+of+cord+blood+derived+mesenchymal+stem+cell+transplantation+into+rat+myocardium.">In vivo bioluminescence imaging of cord blood derived mesenchymal stem cell transplantation into rat myocardium.</a> Ann. Nucl. Med. 20, 165-170 (2006).</li><li>Malstrom, S. E., Tornavaca, O., Meseguer, A., Purchio, A. F., West, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+characterization+and+hormonal+regulation+of+kidney+androgen-regulated+protein+(Kap)-luciferase+transgenic+mice.">The characterization and hormonal regulation of kidney androgen-regulated protein (Kap)-luciferase transgenic mice.</a> Toxicol. Sci. 79, 266-277 (2004).</li><li>Weir, L. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biophotonic+imaging+in+HO-1.luc+transgenic+mice:+real-time+demonstration+of+gender-specific+chloroform+induced+renal+toxicity.">Biophotonic imaging in HO-1.luc transgenic mice: real-time demonstration of gender-specific chloroform induced renal toxicity.</a> Mutat. Res. 574, 67-75 (2005).</li><li>Rajashekara, G., Glover, D. A., Banai, M., O'Callaghan, D., Splitter, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attenuated+bioluminescent+Brucella+melitensis+mutants+GR019+(virB4),+GR024+(galE),+and+GR026+(BMEI1090-BMEI1091)+confer+protection+in+mice.">Attenuated bioluminescent Brucella melitensis mutants GR019 (virB4), GR024 (galE), and GR026 (BMEI1090-BMEI1091) confer protection in mice.</a> Infect. Immun. 74, 2925-2936 (1090).</li><li>Kadurugamuwa, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noninvasive+biophotonic+imaging+for+monitoring+of+catheter-associated+urinary+tract+infections+and+therapy+in+mice.">Noninvasive biophotonic imaging for monitoring of catheter-associated urinary tract infections and therapy in mice.</a> Infect. Immun. 73, 3878-3887 (2005).</li><li>Ryan, P. L., Youngblood, R. C., Harvill, J., Willard, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photonic+monitoring+in+real+time+of+vascular+endothelial+growth+factor+receptor+2+gene+expression+under+relaxin-induced+conditions+in+a+novel+murine+wound+model.">Photonic monitoring in real time of vascular endothelial growth factor receptor 2 gene expression under relaxin-induced conditions in a novel murine wound model.</a> Ann. N.Y. Acad. Sci. 1041, 398-414 (2005).</li><li>Zhu, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+imaging+of+GFAP+expression+after+neuronal+damage+in+mice.">Non-invasive imaging of GFAP expression after neuronal damage in mice.</a> Neurosci. Lett. 367, 210-212 (2004).</li><li>Noiseux, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesenchymal+stem+cells+overexpressing+Akt+dramatically+repair+infarcted+myocardium+and+improve+cardiac+function+despite+infrequent+cellular+fusion+or+differentiation.">Mesenchymal stem cells overexpressing Akt dramatically repair infarcted myocardium and improve cardiac function despite infrequent cellular fusion or differentiation.</a> Mol. Ther. 14, 840-850 (2006).</li><li>Ajiki, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Composite+tissue+transplantation+in+rats:+fusion+of+donor+muscle+to+the+recipient+site.">Composite tissue transplantation in rats: fusion of donor muscle to the recipient site.</a> Transplant Proc. 37, 208-209 (2005).</li><li>Trivedi, P., Hematti, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Derivation+and+immunological+characterization+of+mesenchymal+stromal+cells+from+human+embryonic+stem+cells.">Derivation and immunological characterization of mesenchymal stromal cells from human embryonic stem cells.</a> Exp. Hematol. 36, 350-359 (2008).</li><li>Ogle, B. M., Cascalho, M., Platt, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+implications+of+cell+fusion.">Biological implications of cell fusion.</a> Nat. Rev. Mol. Cell. Biol. 6, 567-575 (2005).</li><li>Collins, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ImageJ+for+microscopy.">ImageJ for microscopy.</a> BioTechniques. 43, 25-30 (2007).</li></ol>

관심 없음 충돌 선언하지 않습니다.

방법은 작은 동물을 포함하여 생물에서 처음으로, 수 여기에 세포 융합의 개별 식별 및 시간적 분석을 설명했다. 접근 방식은 이후 biophotonic 이미지 분석 Cre - LoxP 재조합을 결합한 제품입니다. 접근 방식은 세포 - 세포 융합뿐만 아니라 추적 의무가있다,뿐만 아니라 바이러스 세포 융합 등 바이러스 감염을 추적하기위한 유용 수 있습니다. 이미지 분석은 신속하고 그것은 동시에 이미지를 여러 작?...

<table border="1"><tbody><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 댓글 </td></tr><tr><td> 네온 Transfection 시스템 </td><td> Invitrogen, Carlsbad, CA </td><td> MPK5000 </td><td></td></tr><tr><td> 네온 100 μL 키트 </td><td> Invitrogen, Carlsbad, CA </td><td> MPK10025 </td><td> R과 E 버퍼를 포함 </td></tr><tr><td> - 가상 분말 </td><td> Invitrogen, Carlsbad, CA </td><td> 12000-022 </td><td></td></tr><tr><td> 태아 소 혈청 (FBS) </td><td> Hyclone, 로건 UT </td><td> SH30070.03 </td><td></td></tr><tr><td> B6.C - TG (CMV - cre) 1Cgn / J </td><td> 잭슨 연구소, 바 하버, ME </td><td> 006,054 </td><td></td></tr><tr><td> 10X 트립신 </td><td> 피셔 과학, 숲 잔디, 뉴저지 </tD&gt; <td> MT - 25-054 - CL </td><td></td></tr><tr><td> L - 글루타민 </td><td> 피셔 과학, 숲 잔디, 뉴저지 </td><td> 25030-081 </td><td></td></tr><tr><td> D - Luciferin </td><td> 캘리퍼스 생명 과학, Hopkinton, MA </td><td> 122,796 </td><td></td></tr><tr><td> Xenogen Biophotonic 이미징 시스템 </td><td> 캘리퍼스 생명 과학, Hopkinton, MA </td><td> IVIS 스펙트럼 </td><td></td></tr><tr><td> 나트륨 Biocarbonate </td><td> 시그마 알드리치, 세인트 루이스, 미주리 </td><td> S6014 - 500G </td><td></td></tr><tr><td> 비 필수 아미노산 </td><td> Invitrogen, Carlsbad, CA </td><td> 11140-050 </td><td></td></tr></tbody></table>

a cre-lox p recombination approach for the detection of cell fusion in vivo

퓨즈에 같은 종류의 두 개 이상의 세포의 기능은 골격근, 뼈 및 태반 등 많은 복잡한 장기를 형성하기 위해 진화를 통해 metazoans에서 널리 사용되었습니다. 현대 연구는 같은 종류의 세포 융합 향상된 기능을 수여 보여줍니다. 예를 들어, 태반 퓨즈의 trophoblast 세포가 syncytiotrophoblast을 형성하면 syncytiotrophoblast 더 unfused trophoblasts 1-4보다 모성 - 태아 장벽에 걸쳐 영양소와 호르몬을 수송 수 있습니다. 최근 연구는 다른 종류의 세포 융합 세포의 운명을 직접 보여준다. 융합에 의한 &quot;복귀&quot;또는 세포 운명의 수정은 한 번 세포 배양 시스템에 국한 것으로 생각되었다. 그러나 줄기 세포 이식의 출현은 우리에 의해 발견 LED와 줄기 세포를 다른 생체내의 체세포와 퓨즈 수 있으며 그 융합은 줄기 세포 분화가 5-7 용이하게합니다. 따라서, 세포 융합은 규제 프로세스는 C따라서 세포 생존과 분화를 촉진의 apable 및 개발, 조직의 수리도 질병의 pathogenesis에 대한 중앙의 중요성 수 있습니다. 적절한 기술의 부족)가 1, 세포 융합의 연구는 제한하는 것은 정확하게 시간이 지남에 따라) 트랙 융합 제품을 융합 제품을 식별하고 2. 여기에 우리가 퓨전시 bioluminescence의 유도 (그림 1)를 통해 모두 한계를 해결하기 위해 새로운 접근 방식을 제시,. bioluminescence는 생체내 8-15 높은 감도로 감지할 수 있습니다 우리는 반딧불 루시페라제 인코딩 구조를 활용 (Photinus pyralis) 유전자에 인접한 위치 정지 코돈은 LoxP 시퀀스의 어귀. Cre recombinase 단백질을 표현하는 세포이 유전자 퓨즈를 표현 세포, LoxP 사이트가 죽습하고 정지 신호 루시페라제의 전송을 허용 excised하는 것입니다. 신호가 induci이기 때문에경, 거짓 - 긍정적인 신호의 발병률은 매우 낮은 것입니다. Cre / LoxP 시스템 16, 17를 사용하는 기존의 방법과는 달리, 우리는 &quot;살아있는&quot;감지 신호를 통합하고이를 처음 생체내의 세포 융합의 속도론을 추적할 수있는 기회를 감당할. 접근 방식을 보여주기 위해, ubiquitously Cre recombinase을 표현 쥐가 floxed 중지 코돈의 루시페라제 스트림을 표현하는 구조로 transfected 줄기 세포의 수신자을 역임했습니다. 줄기 세포는 intramyocardial 주입을 통해 이식되었으며 이식 intravital 이미지 분석의 존재를 추적하기 위해 실시 후 시간이지나면서 심장과 주변 조직의 융합 제품. 이러한 접근 방식은 개발, 질병 또는 성인 조직 복구의 모든 단계에서 모든 조직 유형의 세포 융합을 분석 적응 수 있습니다.

Watch this Scientific Journal Video about A Cre-Lox P Recombination Approach for the Detection of Cell Fusion In Vivo at JoVE.com

A Cre-Lox P Recombination Approach for the Detection of Cell Fusion In Vivo

시간이 지남에 따라 생물을 생활에 세포 융합을 추적하는 방법을 설명합니다. 접근 이용 Cre -를<em&gt; LoxP</em&gt; 재조합은 세포 융합에 루시페라제 표현을 유도합니다. 생성되는 발광 신호는 주변 조직의 ~ 1,000 세포의 검출의 감도와 biophotonic 이미징 시스템을 사용하여 살아있는 생물에서 발견하실 수 있습니다.

세포 융합의 탐지를위한 Cre - 훈제 연어 P 재조합 접근법 VIVO에서</em

The ability of two or more cells of the same type to fuse has been utilized in metazoans throughout evolution to form many complex organs, including ...

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Research

JoVE Journal

Bioengineering

A Cre-Lox P Recombination Approach for the Detection of Cell Fusion In Vivo

18.7K 조회수.  University of Wisconsin-Madison. 시간이 지남에 따라 생물을 생활에 세포 융합을 추적하는 방법을 설명합니다. 접근 이용 Cre -를 LoxP 재조합은 세포 융합에 루시페라제 표현을 유도합니다. 생성되는 발광 신호는 주변 조직의 ~ 1,000 세포의 검출의 감도와 biophotonic 이미징 시스템을 사용하여 살아있는 생물에서 발견하실 수 있습니다.

비디오: A Cre-Lox P Recombination Approach for the Detection of Cell Fusion In Vivo

Biomolecular Detection employing the Interferometric Reflectance Imaging Sensor (IRIS)

The sensitive measurement of biomolecular interactions has use in many fields and industries such as basic biology and microbiology, environmental/agricultural/biodefense monitoring, nanobiotechnology, and more. For diagnostic applications, monitoring (detecting) the presence, absence, or abnormal expression of targeted proteomic or genomic biomarkers found in patient samples can be used to determine treatment approaches or therapy efficacy. In the research arena, information on molecular affinities and specificities are useful for fully characterizing the systems under investigation.
Many of the current systems employed to determine molecular concentrations or affinities rely on the use of labels. Examples of these systems include immunoassays such as the enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR) techniques, gel electrophoresis assays, and mass spectrometry (MS). Generally, these labels are fluorescent, radiological, or colorimetric in nature and are directly or indirectly attached to the molecular target of interest. Though the use of labels is widely accepted and has some benefits, there are drawbacks which are stimulating the development of new label-free methods for measuring these interactions. These drawbacks include practical facets such as increased assay cost, reagent lifespan and usability, storage and safety concerns, wasted time and effort in labelling, and variability among the different reagents due to the labelling processes or labels themselves. On a scientific research basis, the use of these labels can also introduce difficulties such as concerns with effects on protein functionality/structure due to the presence of the attached labels and the inability to directly measure the interactions in real time.
Presented here is the use of a new label-free optical biosensor that is amenable to microarray studies, termed the Interferometric Reflectance Imaging Sensor (IRIS), for detecting proteins, DNA, antigenic material, whole pathogens (virions) and other biological material. The IRIS system has been demonstrated to have high sensitivity, precision, and reproducibility for different biomolecular interactions [1-3]. Benefits include multiplex imaging capacity, real time and endpoint measurement capabilities, and other high-throughput attributes such as reduced reagent consumption and a reduction in assay times. Additionally, the IRIS platform is simple to use, requires inexpensive equipment, and utilizes silicon-based solid phase assay components making it compatible with many contemporary surface chemistry approaches.
Here, we present the use of the IRIS system from preparation of probe arrays to incubation and measurement of target binding to analysis of the results in an endpoint format. The model system will be the capture of target antibodies which are specific for human serum albumin (HSA) on HSA-spotted substrates.

Biomolecular Detection employing the Interferometric Reflectance Imaging Sensor IRIS

Quantitative, high-throughput, real-time, and label-free biomolecular detection (DNA, protein, etc.) on SiO2 surfaces can be achieved using a simple interferometric technique which relies on LED illumination, minimal optical components, and a camera. The Interferometric Reflectance Imaging Sensor (IRIS) is inexpensive, simple to use, and amenable to microarray formats.

Interferometric 반사율 이미징 센서 (IRIS)를 채용 Biomolecular 감지

The sensitive measurement of biomolecular interactions has use in many fields and industries such as basic biology and microbiology, ...

연구

생체공학

Fabrication of Electrochemical-DNA Biosensors for the Reagentless Detection of Nucleic Acids, Proteins and Small Molecules

As medicine is currently practiced, doctors send specimens to a central laboratory for testing and thus must wait hours or days to 
receive the results. Many patients would be better served by rapid, bedside tests. To this end our laboratory and others have developed a versatile, reagentless 
biosensor platform that supports the quantitative, reagentless, electrochemical detection of nucleic acids (DNA, RNA), proteins (including antibodies) and small 
molecules analytes directly in unprocessed clinical and environmental samples. In this video, we demonstrate the preparation and use of several biosensors in this 
"E-DNA" class. In particular, we fabricate and demonstrate sensors for the detection of a target DNA sequence in a polymerase chain reaction mixture, an HIV-specific antibody and the drug cocaine. The preparation procedure requires only three hours of hands-on effort followed by an overnight incubation, and their use requires only minutes.

"E-DNA" sensors, reagentless, electrochemical biosensors that perform well even when challenged directly in blood and other complex matrices, have been adapted to the detection of a wide range of nucleic acid, protein and small molecule analytes. Here we present a general procedure for the fabrication and use of such sensors.

핵산, 단백질과 작은 분자의 Reagentless 감지를위한 전기 -의 DNA Biosensors의 제작

As medicine is currently practiced, doctors send specimens to a central laboratory for testing and thus must wait hours or days to 
receive the results. ...

Creating Two-Dimensional Patterned Substrates for Protein and Cell Confinement

Microcontact printing provides a rapid, highly reproducible method for the creation of well-defined patterned substrates.1 While microcontact printing can be employed to directly print a large number of molecules, including proteins,2 DNA,3 and silanes,4 the formation of self-assembled monolayers (SAMs) from long chain alkane thiols on gold provides a simple way to confine proteins and cells to specific patterns containing adhesive and resistant regions. This confinement can be used to control cell morphology and is useful for examining a variety of questions in protein and cell biology. Here, we describe a general method for the creation of well-defined protein patterns for cellular studies.5 This process involves three steps: the production of a patterned master using photolithography, the creation of a PDMS stamp, and microcontact printing of a gold-coated substrate. Once patterned, these cell culture substrates are capable of confining proteins and/or cells (primary cells or cell lines) to the pattern.
The use of self-assembled monolayer chemistry allows for precise control over the patterned protein/cell adhesive regions and non-adhesive regions; this cannot be achieved using direct protein stamping. Hexadecanethiol, the long chain alkane thiol used in the microcontact printing step, produces a hydrophobic surface that readily adsorbs protein from solution. The glycol-terminated thiol, used for backfilling the non-printed regions of the substrate, creates a monolayer that is resistant to protein adsorption and therefore cell growth.6 These thiol monomers produce highly structured monolayers that precisely define regions of the substrate that can support protein adsorption and cell growth. As a result, these substrates are useful for a wide variety of applications from the study of intercellular behavior7 to the creation of microelectronics.8
While other types of monolayer chemistry have been used for cell culture studies, including work from our group using trichlorosilanes to create patterns directly on glass substrates,9 patterned monolayers formed from alkane thiols on gold are straight-forward to prepare. Moreover, the monomers used for monolayer preparation are commercially available, stable, and do not require storage or handling under inert atmosphere. Patterned substrates prepared from alkane thiols can also be recycled and reused several times, maintaining cell confinement.10

Self-assembled monolayers (SAMs) formed from long chain alkane thiols on gold provide well-defined substrates for the formation of protein patterns and cell confinement. Microcontact printing of hexadecanethiol using a polydimethylsiloxane (PDMS) stamp followed by backfilling with a glycol-terminated alkane thiol monomer produces a pattern where protein and cells adsorb only to the stamped hexadecanethiol region.

단백질과 세포 교도소에 대한 2 차원 문양의 기판 만들기

Microcontact printing provides a rapid, highly reproducible method for the creation of well-defined patterned substrates.1  While microcontact printing ...

Simple Microfluidic Devices for in vivo Imaging of C. elegans, Drosophila and Zebrafish

Micro fabricated fluidic devices provide an accessible micro-environment for in vivo studies on small organisms. Simple fabrication processes are available for microfluidic devices using soft lithography techniques 1-3. Microfluidic devices have been used for sub-cellular imaging 4,5, in vivo laser microsurgery 2,6 and cellular imaging 4,7. In vivo imaging requires immobilization of organisms. This has been achieved using suction 5,8, tapered channels 6,7,9, deformable membranes 2-4,10, suction with additional cooling 5, anesthetic gas 11, temperature sensitive gels 12, cyanoacrylate glue 13 and anesthetics such as levamisole 14,15. Commonly used anesthetics influence synaptic transmission 16,17 and are known to have detrimental effects on sub-cellular neuronal transport 4. In this study we demonstrate a membrane based poly-dimethyl-siloxane (PDMS) device that allows anesthetic free immobilization of intact genetic model organisms such as Caenorhabditis elegans (C. elegans), Drosophila larvae and zebrafish larvae. These model organisms are suitable for in vivo studies in microfluidic devices because of their small diameters and optically transparent or translucent bodies. Body diameters range from ~10 &mu;m to ~800 &mu;m for early larval stages of C. elegans and zebrafish larvae and require microfluidic devices of different sizes to achieve complete immobilization for high resolution time-lapse imaging. These organisms are immobilized using pressure applied by compressed nitrogen gas through a liquid column and imaged using an inverted microscope. Animals released from the trap return to normal locomotion within 10 min.
We demonstrate four applications of time-lapse imaging in C. elegans namely, imaging mitochondrial transport in neurons, pre-synaptic vesicle transport in a transport-defective mutant, glutamate receptor transport and Q neuroblast cell division. Data obtained from such movies show that microfluidic immobilization is a useful and accurate means of acquiring in vivo data of cellular and sub-cellular events when compared to anesthetized animals (Figure 1J and 3C-F 4).
Device dimensions were altered to allow time-lapse imaging of different stages of C. elegans, first instar Drosophila larvae and zebrafish larvae. Transport of vesicles marked with synaptotagmin tagged with GFP (syt.eGFP) in sensory neurons shows directed motion of synaptic vesicle markers expressed in cholinergic sensory neurons in intact first instar Drosophila larvae. A similar device has been used to carry out time-lapse imaging of heartbeat in ~30 hr post fertilization (hpf) zebrafish larvae. These data show that the simple devices we have developed can be applied to a variety of model systems to study several cell biological and developmental phenomena in vivo.

Simple Microfluidic Devices for in vivo Imaging of C. elegans, Drosophila and Zebrafish

A simple microfluidic device has been developed to perform anesthetic free in vivo imaging of C. elegans, intact Drosophila larvae and zebrafish larvae. The device utilizes a deformable PDMS membrane to immobilize these model organisms in order to perform time lapse imaging of numerous processes such as heart beat, cell division and sub-cellular neuronal transport. We demonstrate the use of this device and show examples of different types of data collected from different model systems.

에 대한 간단한 마이크로 유체 장치<em&gt; 생체 내</em의&gt; 이미징<em&gt; C. elegans</em&gt;<em&gt; Drosophila</em&gt;와 Zebrafish

Micro fabricated fluidic devices provide an accessible micro-environment for in vivo studies on small organisms. Simple fabrication processes are ...

Microinjection Wound Assay and In vivo Localization of Epidermal Wound Response Reporters in Drosophila Embryos.

The Drosophila embryo develops a robust epidermal layer that serves both to protect the internal cells from a harsh external environment as well as to maintain cellular homeostasis. Puncture injury with glass needles provides a direct method to trigger a rapid epidermal wound response that activates wound transcriptional reporters, which can be visualized by a localized reporter signal in living embryos or larvae. Puncture or laser injury also provides signals that promote the recruitment of hemocytes to the wound site. Surprisingly, severe (through and through) puncture injury in late stage embryos only rarely disrupts normal embryonic development, as greater than 90% of such wounded embryos survive to adulthood when embryos are injected in an oil medium that minimizes immediate leakage of hemolymph from puncture sites. The wound procedure does require micromanipulation of the Drosophila embryos, including manual alignment of the embryos on agar plates and transfer of the aligned embryos to microscope slides. The Drosophila epidermal wound response assay provides a quick system to test the genetic requirements of a variety of biological functions that promote wound healing, as well as a way to screen for potential chemical compounds that promote wound healing. The short life cycle and easy culturing routine make Drosophila a powerful model organism. Drosophila clean wound healing appears to coordinate the epidermal regenerative response, with the innate immune response, in ways that are still under investigation, which provides an excellent system to find conserved regulatory mechanisms common to Drosophila and mammalian epidermal wounding.

Microinjection Wound Assay and In vivo Localization of Epidermal Wound Response Reporters in Drosophila Embryos.

The embryonic epidermis of very late stage Drosophila embryos provides an in vivo system for rapid puncture wound response analysis and can be combined with genetic manipulations or chemical microinjection treatments to advance studies in wound healing for translation into mammalian models.

미세 주입 상처 분석 및<em&gt; 생체 내</em&gt; 표피의 현지화는 응답 기자의 상처<em&gt; 초파리</em&gt; 태아.

The Drosophila embryo develops a robust epidermal layer that serves both to protect the internal cells from a harsh external environment as well as to ...

A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo

Insufficient vascularization is considered to be one of the main factors limiting the clinical success of tissue-engineered constructs. In order to evaluate new strategies that aim at improving vascularization, reliable methods are required to make the in-growth of new blood vessels into bio-artificial scaffolds visible and quantify the results. Over the past couple of years, our group has introduced a full skin defect model that enables the direct visualization of blood vessels by transillumination and provides the possibility of quantification through digital segmentation. In this model, one surgically creates full skin defects in the back of mice and replaces them with the material tested. Molecules or cells of interest can also be incorporated in such materials to study their potential effect. After an observation time of one&#8217;s own choice, materials are explanted for evaluation. Bilateral wounds provide the possibility of making internal comparisons that minimize artifacts among individuals as well as of decreasing the number of animals needed for the study. In comparison to other approaches, our method offers a simple, reliable and cost effective analysis. We have implemented this model as a routine tool to perform high-resolution screening when testing vascularization of different biomaterials and bio-activation approaches.

A Full Skin Defect Model to Evaluate Vascularization of Biomaterials In Vivo

Vascularization is key to approaches in successful tissue engineering. Therefore, reliable technologies are required to evaluate the development of vascular networks in tissue-constructs. Here we present a simple and cost-effective method to visualize and quantify vascularization in vivo.

전체 피부 결함 모델은 생체의 혈관 신생을 평가하기<em&gt; 생체</em

Insufficient vascularization is considered to be one of the main factors limiting the clinical success of tissue-engineered constructs. In order to ...

A Label-free Technique for the Spatio-temporal Imaging of Single Cell Secretions

Inter-cellular communication is an integral part of a complex system that helps in maintaining basic cellular activities. As a result, the malfunctioning of such signaling can lead to many disorders. To understand cell-to-cell signaling, it is essential to study the spatial and temporal nature of the secreted molecules from the cell without disturbing the local environment. Various assays have been developed to study protein secretion, however, these methods are typically based on fluorescent probes which disrupt the relevant signaling pathways. To overcome this limitation, a label-free technique is required.
In this paper, we describe the fabrication and application of a label-free localized surface plasmon resonance imaging (LSPRi) technology capable of detecting protein secretions from a single cell. The plasmonic nanostructures are lithographically patterned onto a standard glass coverslip and can be excited using visible light on commercially available light microscopes. Only a small fraction of the coverslip is covered by the nanostructures and hence this technique is well suited for combining common techniques such as fluorescence and bright-field imaging.
A multidisciplinary approach is used in this protocol which incorporates sensor nanofabrication and subsequent biofunctionalization, binding kinetics characterization of ligand and analyte, the integration of the chip and live cells, and the analysis of the measured signal. As a whole, this technology enables a general label-free approach towards mapping cellular secretions and correlating them with the responses of nearby cells.

Inter-cellular communication is critical for controlling various physiological activities within and outside the cell. This paper describes a protocol for measuring the spatio-temporal nature of single cell secretions. To achieve this, a multidisciplinary approach is used which integrates label-free nanoplasmonic sensing with live cell imaging.

단일 세포 분비물의 공간 - 시간 영상의 레이블이없는 기술

Inter-cellular communication is an integral part of a complex system that helps in maintaining basic cellular activities. As a result, the malfunctioning ...

Hollow Fiber Bioreactors for In Vivo-like Mammalian Tissue Culture

Tissue culture has been used for over 100 years to study cells and responses ex vivo. The convention of this technique is the growth of anchorage dependent cells on the 2-dimensional surface of tissue culture plastic. More recently, there is a growing body of data demonstrating more in vivo-like behaviors of cells grown in 3-dimensional culture systems. This manuscript describes in detail the set-up and operation of a hollow fiber bioreactor system for the in vivo-like culture of mammalian cells. The hollow fiber bioreactor system delivers media to the cells in a manner akin to the delivery of blood through the capillary networks in vivo. The system is designed to fit onto the shelf of a standard CO2 incubator and is simple enough to be set-up by any competent cell biologist with a good understanding of aseptic technique. The systems utility is demonstrated by culturing the hepatocarcinoma cell line HepG2/C3A for 7 days. Further to this and in line with other published reports on the functionality of cells grown in 3-dimensional culture systems the cells are shown to possess increased albumin production (an important hepatic function) when compared to standard 2-dimensional tissue culture.

Hollow Fiber Bioreactors for In Vivo-like Mammalian Tissue Culture

The functional behavior of cells in culture can be improved by culturing in more in vivo-like 3-dimensional culture environments16-21. This manuscript describes the set-up and operation of a hollow fiber bioreactor system for in vivo-like mammalian tissue culture.

에 대한 중공의 생물 반응<em&gt; 생체</em&gt; -처럼 포유류 조직 문화

Tissue culture has been used for over 100 years to study cells and responses ex vivo. The convention of this technique is the growth of anchorage ...

Preparation of 3D Collagen Gels and Microchannels for the Study of 3D Interactions In Vivo

Historically, most cellular processes have been studied in only 2 dimensions. While these studies have been informative about general cell signaling mechanisms, they neglect important cellular cues received from the structural and mechanical properties of the local microenvironment and extracellular matrix (ECM). To understand how cells interact within a physiological ECM, it is important to study them in the context of 3 dimensional assays. Cell migration, cell differentiation, and cell proliferation are only a few processes that have been shown to be impacted by local changes in the mechanical properties of a 3-dimensional ECM. Collagen I, a core fibrillar component of the ECM, is more than a simple structural element of a tissue. Under normal conditions, mechanical cues from the collagen network direct morphogenesis and maintain cellular structures. In diseased microenvironments, such as the tumor microenvironment, the collagen network is often dramatically remodeled, demonstrating altered composition, enhanced deposition and altered fiber organization. In breast cancer, the degree of fiber alignment is important, as an increase in aligned fibers perpendicular to the tumor boundary has been correlated to poorer patient prognosis1. Aligned collagen matrices result in increased dissemination of tumor cells via persistent migration2,3. The following is a simple protocol for embedding cells within a 3-dimensional, fibrillar collagen hydrogel. This protocol is readily adaptable to many platforms, and can reproducibly generate both aligned and random collagen matrices for investigation of cell migration, cell division, and other cellular processes in a tunable, 3-dimensional, physiological microenvironment.

Preparation of 3D Collagen Gels and Microchannels for the Study of 3D Interactions In Vivo

Collagen is a core component of the ECM, and provides essential cues for several cellular processes ranging from migration to differentiation and proliferation. Provided here is a protocol for embedding cells within 3D collagen hydrogels, and a more advanced technique for generating randomized or aligned collagen matrices using PDMS microchannels.

3D 상호 작용의 연구에 대한 3D 콜라겐 젤과 마이크로 채널 준비<em&gt; 생체</em

Historically, most cellular processes have been studied in only 2 dimensions.  While these studies have been informative about general cell signaling ...

Visual Detection of Multiple Nucleic Acids in a Capillary Array

Multi-target, short time, and resource-affordable methodologies for the detection of multiple nucleic acids in a single, easy to operate test are urgently needed in disease diagnosis, microbial monitoring, genetically modified organism (GMO) detection, and forensic analysis. We have previously described the platform called CALM (Capillary Array-based Loop-mediated isothermal amplification for Multiplex visual detection of nucleic acids). Herein, we describe improved fabrication and performance processes for this platform. Here, we apply a small, ready-to-use cassette assembled by capillary array for multiplex visual detection of nucleic acids. The capillary array is pre-treated into a hydrophobic and hydrophilic pattern before fixing loop-mediated isothermal amplification (LAMP) primer sets in capillaries. After assembly of the loading adaptor, LAMP reaction mixture is loaded and isolated into each capillary, due to capillary force by a single pipetting step. The LAMP reactions are performed in parallel in the capillaries. The results are visually read out by illumination with a hand-held UV flashlight. Using this platform, we demonstrate monitoring of 8 frequently appearing elements and genes in GMO samples with high specificity and sensitivity. In summary, the platform described herein is intended to facilitate the detection of multiple nucleic acids. We believe it will be widely applicable in fields where high-throughput nucleic acid analysis is required.

This protocol describes the fabrication of a small, ready-to-use cassette that can be applied for visual detection of multiple nucleic acids in a single, test that is easy to operate. In this approach, a capillary array was used for multiplex and highly efficient detection of GMO targets.

모 세관 배열에 여러 개의 핵 산의 비주얼 검색

Multi-target, short time, and resource-affordable methodologies for the detection of multiple nucleic acids in a single, easy to operate test are urgently ...