Name: ordering single cells and single embryos in 3d confinement: a new device for high content screening
Uploaded: 2016-09-18T13:00:00.000+00:00
Duration: 14 min 22 s
Description: Watch this Scientific Journal Video about Ordering Single Cells and Single Embryos in 3D Confinement: A New Device for High Content Screening at JoVE.com

우리는 안티 Giantin 항체, M. Labouesse 랩으로 우리를 제공하기위한 L. Brino (IGBMC 높은 콘텐츠 상영 시설하여 Illkirch, 프랑스)을 인정합니다. C.에 대한 간스 (IGBMC)와 B Séraphin 연구소. 효모 (IGBMC), 분열 효모 세포에 대한 형광 HeLa 세포 (MPI-CBG, 드레스덴), J. 모슬리 (다트머스 의과 대학)와 JQ 우 (오하이오 주립 대학)에 대한 E. Paluch 및 A. 하이 먼 신진에 대한; A. Hoël과 실험 도움 F. Evenou, 기술 지원을위한 C. 릭 (IBMC, 스트라스부르, 프랑스), 및 미세에 도움 JC Jeannot (펨토 - 일, 프랑스). 이 작품은 CNRS, 스트라스부르 대학, Conectus에서 기금에 의해 지원되었다, 라 파운데이션 부문은 라 공들인 MEDICALE와 스트라스부르의 CI-FRC를 붓는다.

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우리는 공개 할게 없다.

복제 성형은 &#39;eggcups&#39;를 제조하기 위해 사용 하였다. 제조 공정은 클린 룸을 필요로하지 않는다; 연습이 필요할 수 있지만 그것은 쉽고 간단합니다. 특히, PDMS 스탬프을 떼면 고품질 &#39;eggcups&#39;의 넓은 영역을 생성하기 위해 가장 중요한 단계이다. 이 때문에, 특별한주의가이 단계에서 수행되어야한다. 이 단계를 반복하여 실패한 경우, 실란 화 플라즈마 결합 전에 플라즈마 클리너 파라미터...

시험 관내 세포 - 기반 검정에서의 전류 (2D) 이차원이다. 이 구성은 포유 동물 세포에 대한 자연 아니므 한 생리 관련이 없다; 세포 모양, 크기 및 이기종 표현형의 다양성을 보여줍니다. 같은 비행기 및 세포 소기관의 극단적 인 표현형 (특히 스트레스 섬유) 내에서 무질서한 분포로, 심사 응용 프로그램에 적용 할 때 그들은 추가 심각한 한계를 제시한다. 이는 높은 예산이 매년 소비되는 약물 검사에 대한 임상 시험에서 특히 중요하다. 때문에 약물 검사의 초기 단계에서 인공 2D 문화 조건의 동물 모델에 적용 할 때이 약의 대부분은 비록 실패합니다. 또한,이 방법을 이용하여 특정 세포 소기관 적절 통상 관찰의 평면에 수직 한 평면에서 진화 구조 같은 세포 분열 동안 cytokinetic 액토 마이 오신 고리로서 가시화 될 수 없다. 약간새로운 2 차원 분석은 세포 골격 조직에서 상술 한 단점 중요한 통계는 2,3- 관찰되었다 극복하기 위해 제안되었다. 그러나 이러한 분석은 여전히 존재 하나 심각한 제한 : 세포는 세포가 3 차원 구조를 제시 생체 내에서 관찰되는 것과 대조적으로 매우 확산 표현형을 나타낸다. 배양 방법과 관련이 유물은 강화 된 스트레스 섬유 1,4,5으로 비 생리적 기능을 실행할 수 있습니다. 2D는 6,7 환경에 비해 입체 세포 배양 분석은 여러 장점을 제공한다. 그들은 생리 학적 관련성, 그리고 결과에 따라서 의미가있다. 예를 들어, 히드로 겔에 포함 된 세포는 3 차원 형상의 구조를 표시하지만, 그 모폴로지는 또 다른 하나의 셀에 8,9 다르다. 그러나, 그들의 모폴로지는 심사 응용 프로그램을 복잡하게 다른 하나의 셀에서 차이가있다. 또 다른 전략은 단일 포함하는 것입니다미세 공동 10, 11 셀. 셀의 위치, 모양, 극성 및 내부 세포 조직은 정상화 될 수 있습니다. 세포에 3D와 같은 아키텍처를 제공하는 외에, 마이크로 공동 또한 높은 콘텐츠 스크리닝 연구 10,12-14 수 있습니다; 단셀은 마이크로 어레이 및 세포 내 소기관으로 주문할 수 및 진화가 동시에 관찰 할 수있다. 이 규칙은 세포의 낮은 수 더 나은 시간 / 공간 해상도가 좋은 통계를 제공합니다. 유용한 화합물은 안정적으로 쉽게 식별 할 수 있습니다. 본 연구에서는 높은 콘텐츠 스크리닝 응용 10,12,13 대한 제조 및 새로운 3 차원 형상의 단일 세포 배양 시스템의 애플리케이션을 보여준다. 상기 장치는 탄성 중합체 마이크로 공동 (105 공동 / cm 2), 신조어 &#39;eggcups&#39;(EC)의 배열로 구성된다. 이 작품의 크기 및 EC의 총 부피는 개별 NIH3T3과 헬라 세포의 전형적인 볼륨에 최적화되어 있습니다세포 분열시. 원통형 - - 공동의 형태는 활성 프로세스의 시각화를위한 제대로 동양 세포 형태에 선택됩니다. 15,16 커버 슬립 복제 성형 유리에 부착 된 얇은 폴리 디메틸 실록산 (PDMS) 층 상으로 패턴 EC의 배열을 사용한다. 세포를 원심 분리에 의해 EC에 도입된다. 우리는 2D에서 동일한 세포에 비해 여기 관찰 및 3D (EC)의 세포 소기관 (액틴 스트레스 섬유, 골지 장치 및 핵)의 정상화를보고 (평면)의 표면. 우리는 또한 세포의 유사 분열 17시 cytokinetic 액토 마이 오신 링의 폐쇄로 활동 역학 과정의 관찰을보고합니다. 마지막으로, 우리는 신진 효모, 분열 효모와 C.와 같은 단단한 벽, 다른 시스템에서이 방법의 결과를 보여 모델 시스템의 넓은 범위에가는 방법의 적용 가능성을 확인 간스 배아. 우리는 옆에 상세하고 철저한 페이지를 표시순서 rotocol는 제작 및 3D 미세에 대한 &#39;eggcups&#39;을 적용합니다. 우리의 접근 방식은 간단하고 클린 룸을 필요로하지 않습니다. 우리는이 새로운 방법론은 배양 접시의 교체에, 약물 스크리닝 분석 및 맞춤 의학에 특히 흥미로운 일이 될 것으로 예상된다. 마지막으로, 우리의 장치는 암 (18) 또는 기초 연구 (19), 예를 들어, 외부 자극에 세포 응답의 분포를 연구하는데 도움이 될 것입니다.

<table><tbody><tr><td>ddH&lt;sub&gt;2&lt;/sub&gt;0 (ultrapure)</td><td>Millipore</td><td>-</td><td>Use always fresh water.</td></tr><tr><td>Parafilm (plastic film)</td><td>Bemis</td><td>PM-999</td><td>Adhere Parafilm to the lab bench using some water droplets and ensure a perfect surface flatness.</td></tr><tr><td>Photo-mask</td><td>Selba</td><td>-</td><td>&lt;a href=&quot;http://www.selba.ch/&quot; name=&quot;RANGE!D4&quot;&gt;http://www.selba.ch&lt;/a&gt;</td></tr><tr><td>Silicon wafer</td><td>Siltronix</td><td>-</td><td>&lt;a href=&quot;http://www.siltronix.com/&quot;&gt;http://www.siltronix.com/&lt;/a&gt;</td></tr><tr><td>SU-8 photoresist</td><td>MicroChem</td><td>2000 series</td><td>&lt;a href=&quot;http://www.microchem.com/Prod-SU82000.htm&quot;&gt;http://www.microchem.com/Prod-SU82000.htm&amp;nbsp;&lt;/a&gt;</td></tr><tr><td>working in a fumehood is required; check the data sheet from the manufacturer for more information.</td><td></td><td></td><td></td></tr><tr><td>SU-8 developer</td><td>MicroChem</td><td>-</td><td>&lt;a href=&quot;http://microchem.com/Prod-Ancillaries.htm&quot;&gt;http://microchem.com/Prod-Ancillaries.htm&amp;nbsp;&lt;/a&gt;</td></tr><tr><td>working in a fumehood is required; check the data sheet from the manufacturer for more information</td><td></td><td></td><td></td></tr><tr><td>2-propanol</td><td>Sigma-Aldrich</td><td>19030</td><td>&lt;a href=&quot;http://www.sigmaaldrich.com/catalog/product/sial/i9030?lang=en&amp;amp;region=CA&quot;&gt;http://www.sigmaaldrich.com/catalog/product/sial/i9030?lang=en&amp;amp;region=CA&lt;/a&gt;</td></tr><tr><td>Available from multiple companies.</td><td></td><td></td><td></td></tr><tr><td>Sigmacote (siliconizing reagent )</td><td>Sigma-Aldrich</td><td>SL2-25ML</td><td>&lt;a href=&quot;http://www.sigmaaldrich.com/catalog/product/sigma/sl2?lang=fr&amp;amp;region=FR&quot;&gt;http://www.sigmaaldrich.com/catalog/product/sigma/sl2?lang=fr&amp;amp;region=FR&amp;nbsp;&lt;/a&gt;</td></tr><tr><td>harmful, working in a fumehood is required; check the data sheet from the manufacturer for more information.&amp;nbsp;</td><td></td><td></td><td></td></tr><tr><td>Chlorotrimethylsilane (TMCS)</td><td>Sigma-Aldrich</td><td>386529-100ML</td><td>http://www.sigmaaldrich.com/catalog/product/aldrich/386529?lang=fr&amp;amp;region=FR&amp;nbsp;&amp;nbsp;</td></tr><tr><td colspan="4">TMCS produces acute inhalation and dermal toxicity, and is highly flammable (with ignition flashback able to occur across considerable distances), consequently it should be used in a fume cupboard away from sources of ignition</td></tr><tr><td>Nitrile gloves</td><td>Kleenguard</td><td>57372</td><td>&lt;a href=&quot;http://www.kcprofessional.com/products/ppe/hand-gloves/thin-mil-/57372-kleenguard-g10-blue-nitrile-gloves-m&quot;&gt;http://www.kcprofessional.com/products/ppe/hand-gloves/thin-mil-/57372-kleenguard-g10-blue-nitrile-gloves-m&lt;/a&gt;</td></tr><tr><td>Available from multiple companies.</td><td></td><td></td><td></td></tr><tr><td>Glass coverslips #0</td><td>Knittel glass</td><td>KN00010022593</td><td>&lt;a href=&quot;http://www.knittelglass.com/index_e.htm&quot;&gt;http://www.knittelglass.com/index_e.htm&lt;/a&gt;</td></tr><tr><td>Very fragile. Manipulate gently.</td><td></td><td></td><td></td></tr><tr><td>Sharp straight tweezers</td><td>SPI</td><td>0WSSS-XD</td><td>http://www.2spi.com/catalog/tweezers/t/elec7</td></tr><tr><td>50 ml tube</td><td>BD Falcon</td><td>352070</td><td>http://www.bdbiosciences.com/cellculture/tubes/features/index.jsp</td></tr><tr><td>Available from multiple companies.</td><td></td><td></td><td></td></tr><tr><td>PDMS</td><td>Dow Corning</td><td>Sylgard 184 kit</td><td>&lt;a href=&quot;http://www.dowcorning.com/applications/search/default.aspx?R=131EN&quot;&gt;http://www.dowcorning.com/applications/search/default.aspx?R=131EN&amp;nbsp;&lt;/a&gt;</td></tr><tr><td>The package contains both PDMS base and curing agent. Similar elastomers are available from multiple companies.</td><td></td><td></td><td></td></tr><tr><td>Microscope glass slides</td><td>Dutscher</td><td>100001</td><td>&lt;a href=&quot;http://www.dutscher.com/frontoffice/search&quot;&gt;http://www.dutscher.com/frontoffice/search&lt;/a&gt;</td></tr><tr><td>Available from multiple companies.</td><td></td><td></td><td></td></tr><tr><td>DMEM high-glucose medium</td><td>Fisher Scientific</td><td>41965-039</td><td>&lt;a href=&quot;http://www.fishersci.com/ecomm/servlet/Search?LBCID=12301479&amp;amp;keyWord=41965-039&amp;amp;store=Scientific&amp;amp;nav=0&amp;amp;offSet=0&amp;amp;storeId=10652&amp;amp;langId=-1&amp;amp;fromSearchPage=1&amp;amp;searchType=PROD&quot;&gt;http://www.fishersci.com/ecomm/servlet/Search?LBCID=12301479&amp;amp;keyWord=41965-039&amp;amp;store=Scientific&amp;amp;nav=0&amp;amp;offSet=0&amp;amp;storeId=10652&amp;amp;langId=-1&amp;amp;fromSearchPage=1&amp;amp;searchType=PROD&lt;/a&gt;</td></tr><tr><td>Bovine calf serum</td><td>Sigma-Aldrich</td><td>C8056-500ML</td><td>&lt;a href=&quot;http://www.sigmaaldrich.com/catalog/product/sigma/c8056?lang=en&amp;amp;region=CA&quot;&gt;http://www.sigmaaldrich.com/catalog/product/sigma/c8056?lang=en&amp;amp;region=CA&lt;/a&gt;</td></tr><tr><td>0.25% Trypsin-EDTA</td><td>Fisher Scientific</td><td>25200-072</td><td>&lt;a href=&quot;http://www.fishersci.com/ecomm/servlet/Search?keyWord=25200-072&amp;amp;store=Scientific&amp;amp;nav=0&amp;amp;offSet=0&amp;amp;storeId=10652&amp;amp;langId=-1&amp;amp;fromSearchPage=1&amp;amp;searchType=PROD&quot;&gt;http://www.fishersci.com/ecomm/servlet/Search?keyWord=25200-072&amp;amp;store=Scientific&amp;amp;nav=0&amp;amp;offSet=0&amp;amp;storeId=10652&amp;amp;langId=-1&amp;amp;fromSearchPage=1&amp;amp;searchType=PROD&lt;/a&gt;</td></tr><tr><td>PBS 1x</td><td>Fisher Scientific</td><td>14200-067</td><td>&lt;a href=&quot;http://www.fishersci.com/ecomm/servlet/Search?keyWord=14200-067&amp;amp;store=Scientific&amp;amp;nav=0&amp;amp;offSet=0&amp;amp;storeId=10652&amp;amp;langId=-1&amp;amp;fromSearchPage=1&amp;amp;searchType=PROD&quot;&gt;http://www.fishersci.com/ecomm/servlet/Search?keyWord=14200-067&amp;amp;store=Scientific&amp;amp;nav=0&amp;amp;offSet=0&amp;amp;storeId=10652&amp;amp;langId=-1&amp;amp;fromSearchPage=1&amp;amp;searchType=PROD&lt;/a&gt;</td></tr><tr><td>PBS is at 10x and should be diluted to 1x using ddH&lt;sub&gt;2&lt;/sub&gt;O</td><td></td><td></td><td></td></tr><tr><td>L-15 medium</td><td>Fisher Scientific</td><td>21083-027</td><td>&lt;a href=&quot;http://www.fishersci.com/ecomm/servlet/Search?keyWord=21083-027&amp;amp;store=Scientific&amp;amp;nav=0&amp;amp;offSet=0&amp;amp;storeId=10652&amp;amp;langId=-1&amp;amp;fromSearchPage=1&amp;amp;searchType=PROD&quot;&gt;http://www.fishersci.com/ecomm/servlet/Search?keyWord=21083-027&amp;amp;store=Scientific&amp;amp;nav=0&amp;amp;offSet=0&amp;amp;storeId=10652&amp;amp;langId=-1&amp;amp;fromSearchPage=1&amp;amp;searchType=PROD&lt;/a&gt;</td></tr><tr><td>Medium for atmospheres without CO&lt;sub&gt;2&lt;/sub&gt; control</td><td></td><td></td><td></td></tr><tr><td>Fibronectin&amp;nbsp;</td><td>Sigma-Aldrich</td><td>F1141-5MG</td><td>&lt;a href=&quot;http://www.sigmaaldrich.com/catalog/search?interface=All&amp;amp;term=F1141-5MG&amp;amp;lang=en&amp;amp;region=CA&amp;amp;focus=product&amp;amp;N=0+220003048+219853082+219853286&amp;amp;mode=match%20partialmax&quot;&gt;http://www.sigmaaldrich.com/catalog/search?interface=All&amp;amp;term=F1141-5MG&amp;amp;lang=en&amp;amp;region=CA&amp;amp;focus=product&amp;amp;N=0+220003048+219853082+219853286&amp;amp;mode=match%20partialmax&lt;/a&gt;</td></tr><tr><td>Penicillin &amp;amp; Streptomycin</td><td>Fisher Scientific</td><td>15140-122</td><td>&lt;a href=&quot;http://www.fishersci.com/ecomm/servlet/Search?keyWord=15140-122&amp;amp;store=Scientific&amp;amp;nav=0&amp;amp;offSet=0&amp;amp;storeId=10652&amp;amp;langId=-1&amp;amp;fromSearchPage=1&amp;amp;searchType=CHEM&quot;&gt;http://www.fishersci.com/ecomm/servlet/Search?keyWord=15140-122&amp;amp;store=Scientific&amp;amp;nav=0&amp;amp;offSet=0&amp;amp;storeId=10652&amp;amp;langId=-1&amp;amp;fromSearchPage=1&amp;amp;searchType=CHEM&lt;/a&gt;</td></tr><tr><td>Petri dish P35</td><td>Greiner</td><td>627102</td><td>&lt;a href=&quot;http://www.greinerbioone.com/&quot;&gt;http://www.greinerbioone.com/en/row/articles/catalogue/article/144_11/12885/&lt;/a&gt;</td></tr><tr><td>Petri dish P60</td><td>Greiner</td><td>628163</td><td>&lt;a href=&quot;http://www.greinerbioone.com/&quot;&gt;http://www.greinerbioone.com/nl/belgium/articles/catalogue/article/145_8_bl/24872/&lt;/a&gt;</td></tr><tr><td>Petri dish P94</td><td>Greiner</td><td>633179</td><td>&lt;a href=&quot;http://www.greinerbioone.com/&quot;&gt;http://www.greinerbioone.com/nl/belgium/articles/catalogue/article/146_8_bl/24882/&lt;/a&gt;</td></tr><tr><td>Paraformaldehyde 3 %</td><td>Sigma-Aldrich</td><td>P6148-500G</td><td>&lt;a href=&quot;http://www.sigmaaldrich.com/catalog/product/sial/p6148?lang=fr&amp;amp;region=FR&quot;&gt;http://www.sigmaaldrich.com/catalog/product/sial/p6148?lang=fr&amp;amp;region=FR&amp;nbsp;&lt;/a&gt;</td></tr><tr><td>Harmful in-particular for the eyes, working in a fumehood is required; check the data sheet from the manufacturer for more information.</td><td></td><td></td><td></td></tr><tr><td>Triton 0.5 %</td><td>Sigma-Aldrich</td><td>93443-100ML</td><td>&lt;a href=&quot;http://www.sigmaaldrich.com/catalog/search?interface=All&amp;amp;term=93443-100ML&amp;amp;lang=en&amp;amp;region=CA&amp;amp;focus=product&amp;amp;N=0+220003048+219853082+219853286&amp;amp;mode=match%20partialmax&quot;&gt;http://www.sigmaaldrich.com/catalog/search?interface=All&amp;amp;term=93443-100ML&amp;amp;lang=en&amp;amp;region=CA&amp;amp;focus=product&amp;amp;N=0+220003048+219853082+219853286&amp;amp;mode=match%20partialmax&lt;/a&gt;</td></tr><tr><td>Phallodin-Green Fluorescent Alexa Fluor 488</td><td>InVitrogen</td><td>A12379</td><td>&lt;a href=&quot;http://www.lifetechnologies.com/order/catalog/product/A12379?CID=search-a12379&quot;&gt;http://www.lifetechnologies.com/order/catalog/product/A12379?CID=search-a12379&amp;nbsp;&lt;/a&gt;</td></tr><tr><td>dissolve powder in 1.5 ml methanol</td><td></td><td></td><td></td></tr><tr><td>Alexa Fluor 647</td><td>InVitrogen</td><td>A21245</td><td>1:200 dilution in PBS 1x</td></tr><tr><td>rabbit polyclonal anti-Giantin</td><td>Abcam</td><td>ab24586</td><td>1:500 dilution in PBS 1x</td></tr><tr><td>&lt;a href=&quot;http://www.abcam.com/giantin-antibody-ab24586.html&quot;&gt;http://www.abcam.com/giantin-antibody-ab24586.html&amp;nbsp;&lt;/a&gt;</td><td></td><td></td><td></td></tr><tr><td>rabbit anti-anillin</td><td>Courtesy of M. Glotzer, Published in Piekny, A. J. &amp;amp; Glotzer, M. Anillin is a scaffold protein that links RhoA, actin, and myosin during cytokinesis. Current biology 18, 30&amp;ndash;6 (2008).</td><td></td><td>1:500 dilution in PBS 1x</td></tr><tr><td>Anti-phosphotyrosine</td><td>Transduction Lab</td><td>610000</td><td>http://www.bdbiosciences.com/ptProduct.jsp?ccn=610000</td></tr><tr><td>Cy3 goat anti-rabbit&amp;nbsp;</td><td>Jackson Immunoresearch</td><td>111-166-047</td><td>&lt;a href=&quot;http://www.jacksonimmuno.com/catalog/catpages/fab-rab.asp&quot;&gt;http://www.jacksonimmuno.com/catalog/catpages/fab-rab.asp&amp;nbsp;&lt;/a&gt;</td></tr><tr><td>1:1,000 dilution in PBS 1x</td><td></td><td></td><td></td></tr><tr><td>DAPI</td><td>Sigma-Aldrich</td><td>D8417&amp;nbsp;</td><td>&lt;a href=&quot;http://www.sigmaaldrich.com/catalog/product/sigma/d8417?lang=fr&amp;amp;region=FR&quot;&gt;http://www.sigmaaldrich.com/catalog/product/sigma/d8417?lang=fr&amp;amp;region=FR&amp;nbsp;&lt;/a&gt;</td></tr><tr><td>1 mg/ml for 1 min</td><td></td><td></td><td></td></tr><tr><td>Glycerol</td><td>Sigma-Aldrich</td><td>G2025</td><td>&lt;a href=&quot;http://www.sigmaaldrich.com/catalog/search?interface=All&amp;amp;term=G2025&amp;amp;lang=en&amp;amp;region=CA&amp;amp;focus=product&amp;amp;N=0+220003048+219853082+219853286&amp;amp;mode=match%20partialmax&quot;&gt;http://www.sigmaaldrich.com/catalog/search?interface=All&amp;amp;term=G2025&amp;amp;lang=en&amp;amp;region=CA&amp;amp;focus=product&amp;amp;N=0+220003048+219853082+219853286&amp;amp;mode=match%20partialmax&lt;/a&gt;</td></tr><tr><td>Mineral oil</td><td>Sigma-Aldrich</td><td>M8410-500ML</td><td>&lt;a href=&quot;http://www.sigmaaldrich.com/catalog/search?interface=All&amp;amp;term=M8410-500ML&amp;amp;lang=en&amp;amp;region=CA&amp;amp;focus=product&amp;amp;N=0+220003048+219853082+219853286&amp;amp;mode=match%20partialmax&quot;&gt;http://www.sigmaaldrich.com/catalog/search?interface=All&amp;amp;term=M8410-500ML&amp;amp;lang=en&amp;amp;region=CA&amp;amp;focus=product&amp;amp;N=0+220003048+219853082+219853286&amp;amp;mode=match%20partialmax&lt;/a&gt;</td></tr><tr><td>HeLa cells</td><td>-</td><td>-</td><td>Mammalian cells are available from many companies. See also &lt;strong&gt;Table 1&lt;/strong&gt;</td></tr><tr><td>NIH3T3 cells</td><td>ATCC</td><td>-</td><td>Mammalian cells are available from many companies. See also &lt;strong&gt;Table 1&lt;/strong&gt;</td></tr><tr><td>Fission yeast&amp;nbsp;</td><td>-</td><td>-</td><td>For details on strains, contact the corresponding author.&amp;nbsp; See also &lt;strong&gt;Table 1&lt;/strong&gt;</td></tr><tr><td>&lt;em&gt;C. elegans&lt;/em&gt; worms</td><td>-</td><td>-</td><td>For details, contact the corresponding author.&amp;nbsp; See also &lt;strong&gt;Table 1&lt;/strong&gt;</td></tr><tr><td>YES (Agar) + 5 Supplements included</td><td>MP Biomedicals</td><td>4101-732</td><td>&lt;a href=&quot;http://www.mpbio.com/search.php?q=4101-732&amp;amp;s=Search&quot;&gt;http://www.mpbio.com/search.php?q=4101-732&amp;amp;s=Search&amp;nbsp;&lt;/a&gt;</td></tr><tr><td>For preparation: follow&amp;nbsp; instructions as given on the box</td><td></td><td></td><td></td></tr><tr><td>YES (Media) + 5 Supplements included</td><td>MP Biomedicals</td><td>4101-522</td><td>&lt;a href=&quot;http://www.mpbio.com/search.php?q=4101-522&amp;amp;s=Search&quot;&gt;http://www.mpbio.com/search.php?q=4101-522&amp;amp;s=Search&amp;nbsp;&lt;/a&gt;</td></tr><tr><td>For preparation: follow the instructions as given on the box</td><td></td><td></td><td></td></tr><tr><td>EMM (Media)</td><td>MP Biomedicals</td><td>4110-012</td><td>&lt;a href=&quot;http://www.mpbio.com/search.php?q=4110-012&amp;amp;s=Search&quot;&gt;http://www.mpbio.com/search.php?q=4110-012&amp;amp;s=Search&amp;nbsp;&lt;/a&gt;</td></tr><tr><td>For preparation: follow&amp;nbsp; instructions as given on the box</td><td></td><td></td><td></td></tr><tr><td>Filter sterilized EMM (Media) - Only for imaging</td><td>MP Biomedicals</td><td>4110-012</td><td>For preparation: follow instructions as given on the box. Filter sterilize the media using a 0.22 &amp;micro;m filter instead of autoclaving. This gives transparency to the media and reduces the autofluorescence.</td></tr><tr><td>Supplements (for EMM)</td><td>MP Biomedicals</td><td>4104-012</td><td>&lt;a href=&quot;http://www.mpbio.com/search.php?q=4104-012&amp;amp;s=Search&quot;&gt;http://www.mpbio.com/search.php?q=4104-012&amp;amp;s=Search&lt;/a&gt;</td></tr><tr><td>&amp;nbsp;(Add 225 mg/L into the EMM media before autoclaving or filtering)</td><td></td><td></td><td></td></tr><tr><td>Stericup and Steritop Vaccum driven sterile filters</td><td>Millipore</td><td>-</td><td>&lt;a href=&quot;http://www.millipore.com/cellbiology/flx4/cellculture_prepare&amp;amp;tab1=2&amp;amp;tab2=1#tab2=1:tab1=2&quot;&gt;http://www.millipore.com/cellbiology/flx4/cellculture_prepare&amp;amp;tab1=2&amp;amp;tab2=1#tab2=1:tab1=2&lt;/a&gt;</td></tr></tbody></table>

ordering single cells and single embryos in 3d confinement: a new device for high content screening

생물 세포는 일반적으로 평면 (2D) 표면에서 관찰된다. 이 조건은 생리적없고, 표현형 및 모양이 매우 다양하다. 이러한 환경에서 세포 기반 스크리닝 따라서 심각한 한계를 가지고 : 세포 소기관은 이종 및 / 또는 특정 세포 소기관이 제대로 시각화 할 수없는 극단적 인 표현형, 세포의 형태 학적 및 크기를 표시합니다. 또한, 생체 내의 세포는 3 차원 환경에서의 위치; 이 상황에서, 세포는 주로하기 때문에 조직의 주변 세포 외 기질과의 상호 작용의 서로 다른 표현형을 보여줍니다. 표준화 및 세포 기반 검정을위한 생리 학적으로 관련 차원 환경에서 하나의 셀의 순서를 생성하기 위해, 여기에서 3D 관내 세포 배양 장치의 미세 가공 및 애플리케이션을보고한다. 이 장치는 마이크로 공동의 2 차원 배열 (일반적으로 10 5 공동 / cm 2), 단일 세포 또는 배아로 가득 각각 구성되어있다. 셀 포지기, 모양, 극성 및 내부 세포 조직은 3 차원 구조를 보여주는 다음 정규화된다. 우리는 커버 슬립에 부착 된 얇은 폴리 디메틸 실록산 상 &#39;eggcups&#39;(PDMS) 층 패턴 마이크로 공동의 배열을 복제 성형을 사용했다. 캐비티가 접착을 촉진하기 피브로넥틴으로 피복 하였다. 균체를 원심 분리에 의해 삽입 하였다. 충전 비율을 80 %까지 허용하는 각 시스템에 대해 최적화되었다. 배아 세포 생존이 확인되었다. 우리는 핵 및 골지 장치와 같은 세포 소기관의 시각화를 위해이 방법을 적용, 이러한 세포 유사 분열 동안 cytokinetic 반지의 폐쇄 활성 프로세스를 연구. 이 장치는 골지체와 핵 정렬에 대한 cytokinetic 링 폐쇄 및 압축 된 표현형 중에 이러한주기적인 축적과 미오신과 액틴의 불균질성과 같은 새로운 기능의 식별을 허용했다. 우리는 포유류 세포, 분열 효모, 출아 효모, C.하는 방법을 특징으로하는 엘레 w각각의 경우에 i 번째 특정 적응. 마지막으로,이 장치의 특성은 약물 스크리닝 분석 및 맞춤 의학에 특히 흥미합니다.

&#39;eggcups&#39;(EC)는 3 차원 환경에서 배향 배아 세포의 가시화를 허용하는 신규 한 고 함량의 스크리닝 방법이다. 또한, 표준 2 차원 (평면) 배양 물에서 관찰하기 어려운 몇몇 세포 과정은이 새로운 방법에 의해 관찰 할 수있다.도 1a (제 1 전술 한 프로토콜도 참조 EC 미세 가공하기위한 절차의 개요를 나타낸다 ). 상기 방법은도 1B. 간단한 빠르고 효율적인 특수 장비의 필요?...

Watch this Scientific Journal Video about Ordering Single Cells and Single Embryos in 3D Confinement: A New Device for High Content Screening at JoVE.com

Ordering Single Cells and Single Embryos in 3D Confinement: A New Device for High Content Screening

우리는 세포와 배아를 연구하는 장치와 새로운 방법을보고합니다. 단일 세포는 정확하게 마이크로 공동 어레이에 정렬됩니다. 그들의 3D 감금은 생리 학적 조건에서 발생하는 3D 환경을 향한 단계이며 소기관 방향을 수 있습니다. 세포의 모양을 제어함으로써,이 설정은 표준 분석에보고 된 변동성을 최소화 할 수 있습니다.

단일 세포 및 3D 감금 단일 배아 주문 : 하이 콘텐츠 스크리닝을위한 새로운 장치

Biological cells are usually observed on flat (2D) surfaces. This condition is not physiological, and phenotypes and shapes are highly variable. Screening ...

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Research

JoVE Journal

Bioengineering

8.5K 조회수.  Institut de Science et d'Ingénierie Supramoléculaires (ISIS), CNRS and Université de Strasbourg.  우리는 세포와 배아를 연구하는 장치와 새로운 방법을보고합니다. 단일 세포는 정확하게 마이크로 공동 어레이에 정렬됩니다. 그들의 3D 감금은 생리 학적 조건에서 발생하는 3D 환경을 향한 단계이며 소기관 방향을 수 있습니다. 세포의 모양을 제어함으로써,이 설정은 표준 분석에보고 된 변동성을 최소화 할 수 있습니다. 

비디오: Ordering Single Cells and Single Embryos in 3D Confinement: A New Device for High Content Screening

Quantitative Analysis of Protein Expression to Study Lineage Specification in Mouse Preimplantation Embryos

This protocol presents a method to perform quantitative, single-cell in situ analyses of protein expression to study lineage specification in mouse preimplantation embryos. The procedures necessary for embryo collection, immunofluorescence, imaging on a confocal microscope, and image segmentation and analysis are described. This method allows quantitation of the expression of multiple nuclear markers and the spatial (XYZ) coordinates of all cells in the embryo. It takes advantage of MINS, an image segmentation software tool specifically developed for the analysis of confocal images of preimplantation embryos and embryonic stem cell (ESC) colonies. MINS carries out unsupervised nuclear segmentation across the X, Y and Z dimensions, and produces information on cell position in three-dimensional space, as well as nuclear fluorescence levels for all channels with minimal user input. While this protocol has been optimized for the analysis of images of preimplantation stage mouse embryos, it can easily be adapted to the analysis of any other samples exhibiting a good signal-to-noise ratio and where high nuclear density poses a hurdle to image segmentation (e.g., expression analysis of embryonic stem cell (ESC) colonies, differentiating cells in culture, embryos of other species or stages, etc.).

This protocol presents a method to perform quantitative, single-cell in situ analysis of protein expression to study lineage specification in mouse preimplantation embryos. The procedures necessary for collection of blastocysts, whole-mount immunofluorescent detection of proteins, imaging of samples on a confocal microscope, and nuclear segmentation and image analysis are described.

단백질 발현의 정량 분석​​은 마우스 착상 전의 배아에서 리니지 사양을 공부하기

This protocol presents a method to perform quantitative, single-cell in situ analyses of protein expression to study lineage specification in mouse ...

연구

발생학

Isolation and Characterization of Single Cells from Zebrafish Embryos

The zebrafish (Danio rerio) is a powerful model organism to study vertebrate development. Though many aspects of zebrafish embryonic development have been described at the morphological level, little is known about the molecular basis of cellular changes that occur as the organism develops. With recent advancements in microfluidics and multiplexing technologies, it is now possible to characterize gene expression in single cells. This allows for investigation of heterogeneity between individual cells of specific cell populations to identify and classify cell subtypes, characterize intermediate states that occur during cell differentiation, and explore differential cellular responses to stimuli. This study describes a protocol to isolate viable, single cells from zebrafish embryos for high throughput multiplexing assays. This method may be rapidly applied to any zebrafish embryonic cell type with fluorescent markers. An extension of this method may also be used in combination with high throughput sequencing technologies to fully characterize the transcriptome of single cells. As proof of principle, the relative abundance of cardiac differentiation markers was assessed in isolated, single cells derived from nkx2.5 positive cardiac progenitors. By evaluation of gene expression at the single cell level and at a single time point, the data support a model in which cardiac progenitors coexist with differentiating progeny. The method and work flow described here is broadly applicable to the zebrafish research community, requiring only a labeled transgenic fish line and access to microfluidics technologies.

This protocol describes a method for isolating single cells from zebrafish embryos, enriching for cells of interest, capturing zebrafish cells in microfluidic based single cell multiplex systems, and assessing gene expression from single cells.

분리 및 Zebrafish의 배아에서 단일 세포의 특성

The zebrafish (Danio rerio) is a powerful model organism to study vertebrate development. Though many aspects of zebrafish embryonic development have been ...

A Semi-high-throughput Imaging Method and Data Visualization Toolkit to Analyze C. elegans Embryonic Development

C. elegans is the premier system for the systematic analysis of cell fate specification and morphogenetic events during embryonic development. One challenge is that embryogenesis dynamically unfolds over a period of about 13 h; this half day-long timescale has constrained the scope of experiments by limiting the number of embryos that can be imaged. Here, we describe a semi-high-throughput protocol that allows for the simultaneous 3D time-lapse imaging of development in 80&#8211;100 embryos at moderate time resolution, from up to 14 different conditions, in a single overnight run. The protocol is straightforward and can be implemented by any laboratory with access to a microscope with point visiting capacity. The utility of this protocol is demonstrated by using it to image two custom-built strains expressing fluorescent markers optimized to visualize key aspects of germ-layer specification and morphogenesis. To analyze the data, a custom program that crops individual embryos out of a broader field of view in all channels, z-steps, and timepoints and saves the sequences for each embryo into a separate tiff stack was built. The program, which includes a user-friendly graphical user interface (GUI), streamlines data processing by isolating, pre-processing, and uniformly orienting individual embryos in preparation for visualization or automated analysis. Also supplied is an ImageJ macro that compiles individual embryo data into a multi-panel file that displays maximum intensity fluorescence projection and brightfield images for each embryo at each time point. The protocols and tools described herein were validated by using them to characterize embryonic development following knock-down of 40 previously described developmental genes; this analysis visualized previously annotated developmental phenotypes and revealed new ones. In summary, this work details a semi-high-throughput imaging method coupled with a cropping program and ImageJ visualization tool that, when combined with strains expressing informative fluorescent markers, greatly accelerates experiments to analyze embryonic development.

A Semi-high-throughput Imaging Method and Data Visualization Toolkit to Analyze C. elegans Embryonic Development

This work describes a semi-high-throughput protocol that allows simultaneous 3D time-lapse imaging of embryogenesis in 80&#8211;100 C. elegans embryos in a single overnight run. Additionally, image processing and visualization tools are included to streamline data analysis. The combination of these methods with custom reporter strains enables detailed monitoring of embryogenesis.

C. elegans 배아 개발을 분석하는 세미 고처리량 이미징 방법 및 데이터 시각화 툴킷

C. elegans is the premier system for the systematic analysis of cell fate specification and morphogenetic events during embryonic development. One ...

Drosophila Embryo Preparation and Microinjection for Live Cell Microscopy Performed using an Automated High Content Analyzer

Modern approaches in quantitative live cell imaging have become an essential tool for exploring cell biology, by enabling the use of statistics and computational modeling to classify and compare biological processes. Although cell culture model systems are great for high content imaging, high throughput studies of cell morphology suggest that ex vivo cultures are limited in recapitulating the morphological complexity found in cells within living organisms. As such, there is a need for a scalable high throughput model system to image living cells within an intact organism. Described here is a protocol for using a high content image analyzer to simultaneously acquire multiple time-lapse videos of embryonic Drosophila melanogaster development during the syncytial blastoderm stage. The syncytial blastoderm has traditionally served as a great in vivo model for imaging biological events; however, obtaining a significant number of experimental replicates for quantitative and high-throughput approaches has been labor intensive and limited by the imaging of a single embryo per experimental repeat. Presented here is a method to adapt imaging and microinjection approaches to suit a high content imaging system, or any inverted microscope capable of automated multipoint acquisition. This approach enables the simultaneous acquisition of 6-12 embryos, depending on desired acquisition factors, within a single imaging session.

Presented here is a protocol to microinject and simultaneously image multiple Drosophila embryos during embryonic development using a plate-based, high content imager.

자동 고함량 분석기를 사용하여 수행된 살아있는 세포 현미경 검사법을 위한 Drosophila 배아 준비 및 미세 주입

Modern approaches in quantitative live cell imaging have become an essential tool for exploring cell biology, by enabling the use of statistics and ...

Three and Four-Dimensional Visualization and Analysis Approaches to Study Vertebrate Axial Elongation and Segmentation

Somitogenesis is a hallmark of vertebrate embryonic development. For years, researchers have been studying this process in a variety of organisms using a wide range of techniques encompassing ex vivo and in vitro approaches. However, most studies still rely on the analysis of two-dimensional (2D) imaging data, which limits proper evaluation of a developmental process like axial extension and somitogenesis involving highly dynamic interactions in a complex 3D space. Here we describe techniques that allow mouse live imaging acquisition, dataset processing, visualization and analysis in 3D and 4D to study the cells (e.g., neuromesodermal progenitors) involved in these developmental processes. We also provide a step-by-step protocol for optical projection tomography and whole-mount immunofluorescence microscopy in mouse embryos (from sample preparation to image acquisition) and show a pipeline that we developed to process and visualize 3D image data. We extend the use of some of these techniques and highlight specific features of different available software (e.g., Fiji/ImageJ, Drishti, Amira and Imaris) that can be used to improve our current understanding of axial extension and somite formation (e.g., 3D reconstructions). Altogether, the techniques here described emphasize the importance of 3D data visualization and analysis in developmental biology, and might help other researchers to better address 3D and 4D image data in the context of vertebrate axial extension and segmentation. Finally, the work also employs novel tools to facilitate teaching vertebrate embryonic development.

Here, we describe computational tools and methods that allow visualization and analysis of three and four-dimensional image data of mouse embryos in the context of axial elongation and segmentation, obtained by in toto optical projection tomography, and by live imaging and whole-mount immunofluorescence staining using multiphoton microscopy.

척추동물 축 신장 및 세분화를 연구하기 위한 세 가지 및 네 차원 시각화 및 분석 접근법

Somitogenesis is a hallmark of vertebrate embryonic development. For years, researchers have been studying this process in a variety of organisms using a ...

Upright Imaging of Drosophila Embryos

Several well-known morphogenetic gradients and cellular movements occur along the dorsal/ventral axis of the Drosophila embryo. However, the current techniques used to view such processes are somewhat limited. The following protocol describes a new technique for mounting fixed and labeled Drosophila embryos for coronal viewing with confocal imaging. This method consists of embedding embryos between two layers of glycerin jelly mounting media, and imaging jelly strips positioned upright. The first step for sandwiching the embryos is to make a thin bedding of glycerin jelly on a slide. Next, embryos are carefully aligned on this surface and covered with a second layer of jelly. After the second layer is solidified, strips of jelly are cut and flipped upright for imaging. Alternatives are described for visualizing the embryos depending upon the type of microscope stand to be used. Since all cells along the dorsal-ventral axis are imaged within a single confocal Z-plane, our method allows precise measurement and comparison of fluorescent signals without photobleaching or light scattering common to 3D reconstructions of longitudinally mounted embryos.

Upright Imaging of Drosophila Embryos

Here we present a mounting protocol for stained Drosophila embryos in an upright position that allows imaging of cross-sections using Confocal microscopy.

의 입식 이미징 Drosophila 태아

Several well-known morphogenetic gradients and cellular movements occur along the dorsal/ventral axis of the Drosophila embryo. However, the current ...

생물학

Three-dimensional Imaging of Bacterial Cells for Accurate Cellular Representations and Precise Protein Localization

The shape of a bacterium is important for its physiology. Many aspects of cell physiology such as cell motility, predation, and biofilm production can be affected by cell shape. Bacterial cells are three-dimensional (3D) objects, although they are rarely treated as such. Most microscopy techniques result in two-dimensional (2D) images leading to the loss of data pertaining to the actual 3D cell shape and localization of proteins. Certain shape parameters, such as Gaussian curvature (the product of the two principal curvatures), can only be measured in 3D because 2D images do not measure both principal curvatures. Additionally, not all cells lie flat when mounting and 2D imaging of curved cells may not accurately represent the shapes of these cells. Accurately measuring protein localization in 3D can help determine the spatial regulation and function of proteins. A forward convolution technique has been developed that uses the blurring function of the microscope to reconstruct 3D cell shapes and to accurately localize proteins. Here, a protocol for preparing and mounting samples for live cell imaging of bacteria in 3D both to reconstruct an accurate cell shape and to localize proteins is described. The method is based on simple sample preparation, fluorescent image acquisition, and MATLAB-based image processing. Many high-quality fluorescent microscopes can be simply modified to take these measurements. These cell reconstructions are computationally intensive and access to high-throughput computational resources is recommended, although not necessary. This method has been successfully applied to multiple bacterial species and mutants, fluorescent imaging modalities, and microscope manufacturers.

This protocol explains how to prepare and mount bacterial samples for live three-dimensional imaging and how to reconstruct the three-dimensional shape of E. coli from those images.

정확한 세포 표현과 정확한 단백질 국소화를 위한 세균 세포의 3차원 이미징

The shape of a bacterium is important for its physiology. Many aspects of cell physiology such as cell motility, predation, and biofilm production can be ...

면역학 및 감염병학

A Manual Small Molecule Screen Approaching High-throughput Using Zebrafish Embryos

Zebrafish have become a widely used model organism to investigate the mechanisms that underlie developmental biology and to study human disease pathology due to their considerable degree of genetic conservation with humans. Chemical genetics entails testing the effect that small molecules have on a biological process and is becoming a popular translational research method to identify therapeutic compounds. Zebrafish are specifically appealing to use for chemical genetics because of their ability to produce large clutches of transparent embryos, which are externally fertilized. Furthermore, zebrafish embryos can be easily drug treated by the simple addition of a compound to the embryo media. Using whole-mount in situ hybridization (WISH), mRNA expression can be clearly visualized within zebrafish embryos. Together, using chemical genetics and WISH, the zebrafish becomes a potent whole organism context in which to determine the cellular and physiological effects of small molecules. Innovative advances have been made in technologies that utilize machine-based screening procedures, however for many labs such options are not accessible or remain cost-prohibitive. The protocol described here explains how to execute a manual high-throughput chemical genetic screen that requires basic resources and can be accomplished by a single individual or small team in an efficient period of time. Thus, this protocol provides a feasible strategy that can be implemented by research groups to perform chemical genetics in zebrafish, which can be useful for gaining fundamental insights into developmental processes, disease mechanisms, and to identify novel compounds and signaling pathways that have medically relevant applications.

The zebrafish is an excellent experimental organism to study vertebrate developmental processes and model human disease. Here, we describe a protocol on how to perform a manual high-throughput chemical screen in zebrafish embryos with a whole-mount in situ hybridization (WISH) read-out.

Zebrafish의 배아를 사용하여 높은 처리량 접근 수동 작은 분자 화면

Zebrafish have become a widely used model organism to investigate the mechanisms that underlie developmental biology and to study human disease pathology ...

Discrimination and Characterization of Heterocellular Populations Using Quantitative Imaging Techniques

Cellular processes are complex and result from the interplay between multiple cell types and their environment. Existing cell biology techniques often do not allow for accurate interpretation of this interplay. Using a quantitative imaging-based approach, we present a high-content protocol for characterizing the dynamic phenotypic responses (i.e. morphology changes, proliferation, apoptosis) of heterogeneous cell populations to changes in environmental stimuli. We highlight our ability to distinguish between cell types based upon either fluorescence intensity or inherent morphology features depending on the application. This platform allows for a more comprehensive characterization of subpopulation response to perturbation while utilizing shorter time, smaller amounts of reagents, and lower likelihood of error than traditional cell biology assays. However, in some cases, cell populations may be difficult to identify and quantitate based on complex cellular features and will require additional troubleshooting; we highlight some of these circumstances in the protocol. We demonstrate this application using response to drug in a cancer model; however, it can easily be applied more broadly to other physiological processes. This protocol allows one to identify subpopulations within a co-culture system and characterize the particular response of each to external stimuli.

We have developed a novel protocol for studying heterocellular population dynamics in response to perturbations. This manuscript describes an imaging-based platform that produces quantitative datasets for simultaneous characterization of multiple cellular phenotypes of heterocellular populations in a robust manner.

정량적 이미징 기법을 이용한 이종 세포 집단의 차별화 및 특성 규명

Cellular processes are complex and result from the interplay between multiple cell types and their environment. Existing cell biology techniques often do ...

Single-Cell Resolution Three-Dimensional Imaging of Intact Organoids

Organoid technology, in vitro 3D culturing of miniature tissue, has opened a new experimental window for cellular processes that govern organ development and function as well as disease. Fluorescence microscopy has played a major role in characterizing their cellular composition in detail and demonstrating their similarity to the tissue they originate from. In this article, we present a comprehensive protocol for high-resolution 3D imaging of whole organoids upon immunofluorescent labeling. This method is widely applicable for imaging of organoids differing in origin, size and shape. Thus far we have applied the method to airway, colon, kidney, and liver organoids derived from healthy human tissue, as well as human breast tumor organoids and mouse mammary gland organoids. We use an optical clearing agent, FUnGI, which enables the acquisition of whole 3D organoids with the opportunity for single-cell quantification of markers. This three-day protocol from organoid harvesting to image analysis is optimized for 3D imaging using confocal microscopy.

The entire 3D structure and cellular content of organoids, as well as their phenotypic resemblance to the original tissue can be captured using the single-cell resolution 3D imaging protocol described here. This protocol can be applied to a wide range of organoids varying in origin, size and shape.

단세포 분해능 3차원 알래기 알코노이드

Organoid technology, in vitro 3D culturing of miniature tissue, has opened a new experimental window for cellular processes that govern organ development ...

단일 세포 및 3D 감금 단일 배아 주문 : 하이 콘텐츠 스크리닝을위한 새로운 장치

요약

더 많은 비디오 탐색

이 비디오의 챕터

단백질 발현의 정량 분석은 마우스 착상 전의 배아에서 리니지 사양을 공부하기

분리 및 Zebrafish의 배아에서 단일 세포의 특성

C. elegans 배아 개발을 분석하는 세미 고처리량 이미징 방법 및 데이터 시각화 툴킷

자동 고함량 분석기를 사용하여 수행된 살아있는 세포 현미경 검사법을 위한 Drosophila 배아 준비 및 미세 주입

척추동물 축 신장 및 세분화를 연구하기 위한 세 가지 및 네 차원 시각화 및 분석 접근법

의 입식 이미징 Drosophila 태아

정확한 세포 표현과 정확한 단백질 국소화를 위한 세균 세포의 3차원 이미징

Zebrafish의 배아를 사용하여 높은 처리량 접근 수동 작은 분자 화면

정량적 이미징 기법을 이용한 이종 세포 집단의 차별화 및 특성 규명

단세포 분해능 3차원 알래기 알코노이드

단일 세포 및 3D 감금 단일 배아 주문 : 하이 콘텐츠 스크리닝을위한 새로운 장치

요약

더 많은 비디오 탐색

이 비디오의 챕터

단백질 발현의 정량 분석​​은 마우스 착상 전의 배아에서 리니지 사양을 공부하기

분리 및 Zebrafish의 배아에서 단일 세포의 특성

C. elegans 배아 개발을 분석하는 세미 고처리량 이미징 방법 및 데이터 시각화 툴킷

자동 고함량 분석기를 사용하여 수행된 살아있는 세포 현미경 검사법을 위한 Drosophila 배아 준비 및 미세 주입

척추동물 축 신장 및 세분화를 연구하기 위한 세 가지 및 네 차원 시각화 및 분석 접근법

의 입식 이미징 Drosophila 태아

정확한 세포 표현과 정확한 단백질 국소화를 위한 세균 세포의 3차원 이미징

Zebrafish의 배아를 사용하여 높은 처리량 접근 수동 작은 분자 화면

정량적 이미징 기법을 이용한 이종 세포 집단의 차별화 및 특성 규명

단세포 분해능 3차원 알래기 알코노이드

단백질 발현의 정량 분석은 마우스 착상 전의 배아에서 리니지 사양을 공부하기