이 작품은 노인학 수상 및 NIH 그랜트 MKGS에 1R01AG031108 - 01 NIH 연수 부여 P30AG013280에서 지원하는에있는 글렌 / 멀리 획기적인 지원했다. MK는 노화에 엘리슨 의료 재단 신규 장학생입니다.

1 부 : 준비 선충류의 성장 미디어 (NGM) 접시 이 섹션은 수명 실험에 사용하기 위해 고체 NGM 플레이트의 준비에 대해 설명합니다. 암피실린 (앰프)와 NGM에 추가 fluorodeoxyuridine (FUDR)를 모두 가지고 아무런 첨가제 및 앰프 / FUDR 접시를 포함하지 표준 NGM 번호판, : 기본 수명 실험은 접시 두 가지 유형이 필요합니다. 암피실린 외국 세균 오염을 방지하는 데 사용됩니다. FUDR은 세포 분열을 억제 계란 생산을 감소하고, 부화에서 계란을 방지할 수 있습니다. 장수 분석을위한 FUDR의 사용은 성인 수명에 영향을 성장 유충에서 분리하기 위해 모든 며칠 벌레를 전송하는 필요를 제거하지 않습니다. 접시의 두 종류 E. 함께 씨앗 아르 이후 UV에 노출에 의해 살해 대장균 박테리아 OP50. <ol><li> NGM (조리법 및 스토리지 노트 섹션 5 참조)와 레이블 배양 접시를 준비합니다. 당신은 NGM 10 ML 당 1 60mm 페트리 접시가 필요합니다. 당신이 이전에 준비 고체 NGM 시작하는 경우, 1.2 단계로 진행합니다. 당신은 갓 autoclaved NGM 시작하는 경우, 1.4 단계로 건너 뜁니다. </li><li> 완전히 30초 펄스의 높이 microwaving하여 고체 NGM을 용융. 최대 건물에서 가압 가스 거품을 방지하기 위해 펄스 사이의 소용돌이 미디어. </li><li> 55 ° C 물 목욕이나 냉각 벤치에 자리 액체 NGM. </li><li> 일단 NGM 55에 도달 ° C 150 MM의 FUDR 33 μL과 100 MG / ML 100 ML NGM 당 암피실린 (앰프 / FUDR 접시)와 섞어 소용돌이 100 μL를 추가합니다. 추가 약물없이 NGM 접시에 대한 1.5 단계로 바로 진행합니다. </li><li> 각 60mm 배양 플레이트에 살균 기법, 피펫 10 ML NGM을 사용합니다. 가능하면 거품을 형성하지 마십시오. 거품이 양식을 할 경우, 그들은 피펫 팁을 사용하여 쐈고 수 있습니다. </li><li> NGM은 응고 및 건조 수 있도록 뚜껑에있는 벤치에 접시를 남겨주세요. 가능하면, 박테리아를 추가하기 전에 2 일간 벤치에 번호판을 떠나지만, 접시가 빨리 필요하면 일일 작동합니다. </li><li> 당일 번호판 E.의 신선한 행진에서 하나의 식민지로 건조, 종자 액체 LB 문화를 완료하기 전에 LB 한천에서 대장균 박테리아를 OP50. 당신은 60mm 판 당 하룻밤 OP50 문화의 최소한 1 ML 준비를해야합니다. </li><li> 37 장소 OP50 문화 박테리아 (문화 포화에 도달한다) 하룻밤 성장 ° C 뿌리와이 가능하다는 특징을 가지고있다. </li><li> 펠렛 OP50 10 분 3,500그램에서 회전에 의해 박테리아. </li><li> 문화 10X 집중 뜨는 및 resuspend 박테리아의 90 %를 제거합니다. </li><li> 확정 NGM 플레이트의 중심에 10X 집중 OP50 문화의 피펫 100 μL. (이상적으로 박테리아가 판 벽 근처에 오는없이 NGM의 중심 영역을 커버한다) 필요할 경우 소용돌이의 번호판은 부드럽게 박테리아 문화를 확산합니다. 미디어의 표면에 결함이 NGM로 버로우로 벌레를 허용 등 NGM의 표면에 피펫 팁을 만지는 피하기 위해보십시오. </li><li> 박테리아 문화가 고체 NGM (일반적으로 약 24 시간 소요)에 건조 수 있도록 밤새 벤치에 접시를 남겨주세요. </li><li> 일단 건조하고 박테리아의 성장을 체포하는 데 충분한 UV 복용으로 접시의 표면을 쉽게받을 수 있습니다. 당신은 Stratagene의 자외선 Stratalinker 2400을 사용하는 경우 : <ul><li> 플레이스 Stratalinker에 접시와 뚜껑을 제거 </li><li> 문을 닫고 Stratalinker 켜십시오 </li><li> 보도 자료 &#39;에너지&#39; </li><li> &#39;9999 &#39;은 키패드를 사용하여 입력 </li><li> 누르면 &#39;시작&#39; </li><li> 플레이트는 약 5 분 동안 자외선에 노출됩니다 </li></ul></li><li> UV - 살해 박테리아와 플레이트는 ° C 최대 1 개월 4에서 거꾸로 저장할 수 있습니다. </li></ol> 2 부 : 동물의 연령 동기 인구를 얻기 누워있는 시간이 초과되었습니다 달걀을 수행 이 섹션에서 우리는 공통의 해치 - 날짜와 웜의 인구를 생성합니다. 이것은 reproductively 활성 성인 시간 정의된 기간 동안 접시에 알을 낳을 수 있도록 이들 계란 개발함으로써 수행됩니다. <ol><li> (선택 사항) FUDR없이 신선한 NGM 접시에 약 20 젊은 성인 웜을 전송합니다. ° C 웜은 세균성 음식의 모든을 통해 전파하고 먹을 수 있도록 25 접시를 남겨주세요. 세균성 음식 섭취 후에는 새로 부화 벌레는 dauer 단계를 체포 성장을 입력합니다. 당신은 처음 접시로 전송 얼마나 많은 벌레에 따라 일주일 이내 주변 dauer의 유충을 볼 시작합니다. Dauer의 유충은 최대 1 개월 다음 단계에 사용할 수 있습니다. </li><li> FUDR없이 신선한 NGM 접시에 (옵션) 전송 20-30 dauer의 유충. 식품 dauer의 유충의 존재 2 일 이내에 reproductively 활성 성인이 될 것이며, 25 이후 며칠 동안 reproductively 활성 상태로 유지 ° C.가 </li><li> FUDR없이 신선한 NGM 접시에 10-15 reproductively 적극적인 성인을 전송합니다. 이 번호판은 시간 초과 에그 누워 (전화) 번호판이라고합니다. </li><li> ° C 6 시간 t를 허용하기 위해 25 TEL 접시를 남겨주세요그는 벌레 시간은 알을 낳기 위해. </li><li> TEL 플레이트에서 어른 벌레를 제거합니다. 플레이트는 시각 성인 제거하기 전에 달걀에 대한 검사를하실 수 있습니다. 벌레가 알을 충분한 숫자를 마련하지 않은 경우 TEL 판은 어른 벌레를 제거하기 전에 24 시간 총 최대 25 ° C에서 남아있을 수 있습니다. </li><li> 20 장소 TEL 판 ° C 계란이 부화하고 벌레가 L4 애벌레 단계 (이것은 일반적으로 야생 유형 C. 엘레간스 2 일이 소요하지만, 속도가 느린 개발 phenotypes와 종자에 대한 더 오래 걸릴 수 있습니다)을 개발하기 전까지. </li></ol> 파트 3 : 수명에 대한 점수 동물 그들은 죽을 때까지이 섹션에서 우리는 2 부에서 벌레의 시대 - 동기 인구를 수행합니다. 웜가 계란 생산 및 박테리아 오염을 방지하기 위해 앰프 / FUDR 접시에 유지되며 외부 자극에 반응하지 않는 경우 죽은 것으로 간주됩니다. <ol><li> 씨앗 앰프 / FUDR 접시에 L4 유충을 전송합니다. 각각의 변형이나 조건이 테스트되는 경우, 그것은 접시마다 25-30 벌레와 2-3 접시를 설정하는 것이 일반적입니다. C. 이미지 엘레간스 생활 단계는 참조 (그림 1)를 제공합니다 1. </li><li> 20 플레이스 앰프 / FUDR 접시 ° 24 시간 동안 C. </li><li> 24 시간 후에, 시각 웜, 미디어, 그리고 박테리아를 평가. 다음 중 하나를 관찰하는 경우 신선한 앰프 / FUDR 접시에 웜을 전송 : <ul><li> 웜는 세균 음식의 대부분을 먹었. 조기 수명의 실험 웜을 1로 2 배에게 고갈되는 세균을 방지하기 위해 일주일을 전송해야합니다. </li><li> 애벌레의 상당수가 존재하고 있습니다. 이것은 일반적으로 벌레가 젊은 성인 대신 L4s로 앰프 / FUDR 접시에 전송하고 FUDR이 효과를 데려가기 전에 몇 알을 낳을 수 있었던 것을 나타냅니다. FUDR은 일반적으로 전체 성인으로 성장의 애벌레를 방지할 수 있지만, 가끔 몇 어른으로 성장하고 실험 동물과 혼동이 될 것입니다. </li><li> 라이브 / 성장 세균이 관찰됩니다. 일반적으로 앰프와 UV - 살인의 조합은 사는 박테리아가 실험을 오염하지 않도록되지만 종종 발생합니다. </li><li> 곰팡이 성장은 미디어에 관찰됩니다. 초기 충분히 잡는다면, 곰팡이 성장은 일반적으로 피펫 팁 또는 주걱을 사용하여 잘라하실 수 있습니다. 일단 그것은 직경이 새 접시에 벌레를 전송하는 일반적으로 쉽게 몇 mm보다 큰 수 자랍니다. </li><li> 절개 범위를 사용하여, 각각의 벌레가 살아 있거나 죽어 있는지 확인합니다. </li><li> 부드럽게 접시를 누릅니다. 그것은 감​​청에 대한 응답으로 이동하면 웜이가 살아있다는 의미이다. </li><li> 웜은 머리 지역에있는 판, 줌 감청에 응답하지 않는 경우. </li><li> 부드럽게 플래티넘 전송 선택으로 벌레의 머리를 누릅니다. 그것의 머리를 이동하여 응답하지 않으면 죽은 벌레를 점수. </li><li> 죽은 벌레는 접시에서 제거할 수 있습니다. </li></ul></li><li> 살아 죽은 날짜와 벌레의 수를 기록합니다. </li><li> 20 ° C.에 접시로 돌아가기 </li><li> 모든 벌레가 죽었을 때까지 반복 3.3 3.6를 통해 모든 2~3일 단계를 반복합니다. </li></ol> 4 부 : 대표 결과. 선충류의 수명 실험에 의해 만들어진 원시 데이터 테스트 각 변형에 대한 살아있는 죽은 벌레의 해당 숫자와 날짜의 목록입니다. 매일 죽는 벌레의 수는 일반적으로 생존 곡선 (그림 2B로 그래픽 꾸몄다는 매일 살아있는 벌레의 비율 (그림 2A), 계산하기 위해 거꾸로되며 시간 초과 에그 누워 당일 것은 일 0로 간주됩니다 ). 연구의 각 웜에 대한 수명은 매일 죽는 벌레의 개수의 계산과 변종 사이의 비교에 대한 의미와 중간 수명을 계산하는 데 사용할 수 있습니다. 매일 살아있는 벌레의 수를 셀 수명 분석에 직접 사용되지 않습니다. 고체 미디어에 유지 벌레는 때로는 &#39;도망&#39;또는 언론 아래 접시 또는 아래의 벽까지 중 크롤 링합니다. 매일 살아있는 벌레의 수는 얼마나 많은 벌레들이 실험 과정을 통해 도망 결정하는 데 사용할 수 있습니다. 도망 웜은 일반적으로 분석에서 제거됩니다. N2, C.위한 벤치 마크로서, 전형적인 중간 수명 20 UV - 살해 박테리아에서 유지 엘레간스 야생 형 변형, ° C는 난자에서 측정한 약 25 일입니다. 제 5 부 : 솔루션 <table border="0"><tbody><tr><td colspan="2"> 선충류 성장 미디어 (NGM), 100 ML : </td></tr><tr><td colspan="2"> 컴바인 : </td></tr><tr><td> 0.3 g </td><td> NaCl </td></tr><tr><td> 0.25 g </td><td> 펩톤 </td></tr><tr><td> 2g </td><td> 한천 </td></tr><tr><td colspan="2"> 40 분 압력솥과 55 식지는 ° C, 다음 추가 : </td></tr><tr><td> 100 μL </td><td> 1 M MgSO 4 </td></tr><tr><td> 100 μL </td><td> 5 MG / ML 콜레스테롤 </td></tr><tr><td> 100 μL </td><td> 1 M CaCl2 </td></tr><tr><td> 1.625 ML </td><td> 1.5 M KPI의 산도 6.0 </td></tr><tr><td colspan="2"> 액체 NGM는 실온에서 번호판을 부어 즉시 사용되거나 응고 허용 및 장기 저장할 수 있습니다 </td></tr></tbody></table><table border="0"><tbody><tr><td colspan="2"> Luria 맛을 (LB), 1L : </td></tr><tr><td> 10g </td><td> BactoTryptone </td></tr><tr><td> 5g </td><td> 효모 추출 </td></tr><tr><td> 10g </td><td> NaCl </td></tr><tr><td> 10 ML </td><td> 1 M 트리스 산도 8.0 </td></tr><tr><td> 1 L </td><td> 탈이온수 </td></tr><tr><td colspan="2"> 압력솥과 실온에서 저장합니다. </td></tr></tbody></table><table border="0"><tbody><tr><td colspan="2"> 1 M MgSO 4, 300 ML : </td></tr><tr><td> 73.95 g </td><td> MgSO 4 </td></tr><tr><td> 300 ML </td><td> 탈이온수 </td></tr><tr><td colspan="2"> 압력솥과 실온에서 저장합니다. </td></tr></tbody></table><table border="0"><tbody><tr><td colspan="2"> 5 MG / ML 콜레스테롤, 200 ML : </td></tr><tr><td> 1g </td><td> 콜레스테롤 </td></tr><tr><td> 200 ML </td><td> 100 % 에탄올 </td></tr><tr><td colspan="2"> 소독 필터 및 실내 온도에 저장합니다. </td></tr></tbody></table><table border="0"><tbody><tr><td colspan="2"> 1 M CaCl 2, 500 ML : </td></tr><tr><td> 27.75 g </td><td> CaCl 2 </td></tr><tr><td> 500 ML </td><td> 탈이온수 </td></tr><tr><td colspan="2"> 소독 필터 및 실내 온도에 저장합니다. </td></tr></tbody></table><table border="0"><tbody><tr><td colspan="2"> 1.5 M KPI 산도 6.0 1L : </td></tr><tr><td colspan="2"> 컴바인 : </td></tr><tr><td> 31.4 g </td><td> KPI 이염 </td></tr><tr><td> 179.6 g </td><td> KPI 일염기의 </td></tr><tr><td> 850 ML </td><td> 탈이온수 </td></tr><tr><td colspan="2"> 히트 KPI는 솔루션으로 분해하도록 혼합하는 동안. 10 N NaOH와 6.0 산도를 조정합니다. </td></tr><tr><td colspan="2"> 1 L.에 탈이온수 추가 </td></tr><tr><td colspan="2"> 압력솥과 실온에서 저장합니다. </td></tr></tbody></table><table border="0"><tbody><tr><td colspan="2"> 1 M 트리스, 산도 8.0 : </td></tr><tr><td> 60.57 g </td><td> 트리스 </td></tr><tr><td> 400 ML </td><td> 탈이온수 </td></tr><tr><td colspan="2"> HCL과 8.0 산도를 조정합니다. </td></tr><tr><td colspan="2"> 500 ML에 탈이온수를 추가합니다. </td></tr><tr><td colspan="2"> 소독 필터 및 실내 온도에 저장합니다. </td></tr></tbody></table><table border="0"><tbody><tr><td colspan="2"> 150 MM Fluorodeoxyuridine (FUDR), 10 ML : </td></tr><tr><td> 0.3693 g </td><td> FUDR </td></tr><tr><td> 10 ML </td><td> 무균 탈이온수 </td></tr><tr><td colspan="2"> -20 ° C.에 저장 </td></tr></tbody></table><table border="0"><tbody><tr><td colspan="2"> 100 MG / ML 암피실린 (앰프), 10 ML : </td></tr><tr><td> 1g </td><td> 암피실린 </td></tr><tr><td> 10 ML </td><td> 무균 탈이온수 </td></tr><tr><td colspan="2"> -20 ° C.에 저장 </td></tr></tbody></table><table border="0"><tbody><tr><td colspan="2"> 50 MG / ML Carbenicillin (수화물), 10 ML : </td></tr><tr><td> 500 MG </td><td> Carbenicillin </td></tr><tr><td> 10 ML </td><td> 무균 탈이온수 </td></tr><tr><td colspan="2"> -20 ° C.에 저장 </td></tr></tbody></table><table border="0"><tbody><tr><td colspan="2"> 1 M 이소 프로필 β - D - Thiogalactopyranoside (IPTG), 10 ML : </td></tr><tr><td> 2.38 g </td><td> IPTG </td></tr><tr><td> 10 ML </td><td> 무균 탈이온수 </td></tr><tr><td colspan="2"> -20 ° C.에 저장 </td></tr></tbody></table><img src="/files/ftp_upload/1152/Figure%201.png" alt="그림 1" /> C. 그림 1. 명시야 이미지 달걀을 포함 엘레간스 생활 단계, 네 개의 애벌레 단계 (L1 - L4) 및 성인. 모든 패널은 성인 남성 (우드 (1988)에서 이미지) 보여주는, 오른쪽 아래를 제외하고 나온것을 보여줍니다. <img src="/files/ftp_upload/1152/Figure%202.png" alt="그림 2" /> C.에서 그림 2. 대표 결과 엘레간스 수명 실험은 DAF - 2 유전자에 돌연변이 포함된 변형으로 야생 유형 변형 N2를 비교. (A) 수집 벌레가 알을 이후 일수를 포함하여 데이터, 매일 관찰 죽은 벌레의 수를, 그리고 매일에 살아 남아있는 원래의 샘플의 비율 (죽은 벌레의 일상 건의부터 계산) 보여주는 표 . (B) 생존 (A)에서 제공하는 수명 데이터에 해당하는 곡선. <imgSRC = &quot;/ files/ftp_upload/1152/Figure 3.jpg&quot;고도 = &quot;그림 3&quot;/&gt; 그림 3. 젊은이와 노인 성인 C. 사이 lipofuscin 대표 비교 엘레간스. 명시야 이미지는 오른쪽 왼쪽 채널 DAPI 형광 이미지에 표시됩니다. 상단 패널은 사일 오래된 웜 및 바닥 패널 (같은 난자에서 측정) 11일 오래된 벌레를 보여 보여줍니다.

<ol>
	<li>Wood, W. B., Wood, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Nematode Caenorhabditis elegans. , 1-16 (1988).</li><li>Sutphin, G. L., Kaeberlein, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dietary+restriction+by+bacterial+deprivation+increases+life+span+in+wild-derived+nematodes.">Dietary restriction by bacterial deprivation increases life span in wild-derived nematodes.</a> Exp Gerontol. 43, 130-135 (2008).</li><li>Garigan, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+analysis+of+tissue+aging+in+Caenorhabditis+elegans:+a+role+for+heat-shock+factor+and+bacterial+proliferation.">Genetic analysis of tissue aging in Caenorhabditis elegans: a role for heat-shock factor and bacterial proliferation.</a> Genetics. 161, 1101-1112 (2002).</li><li>Chen, D., Pan, K. Z., Palter, J. E., Kapahi, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Longevity+determined+by+developmental+arrest+genes+in+Caenorhabditis+elegans.">Longevity determined by developmental arrest genes in Caenorhabditis elegans.</a> Aging Cell. 6, 525-523 (2007).</li><li>Curran, S. P., Ruvkun, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lifespan+regulation+by+evolutionarily+conserved+genes+essential+for+viability.">Lifespan regulation by evolutionarily conserved genes essential for viability.</a> PLoS Genet. 3, e56-e56 (2007).</li><li>Dillin, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rates+of+behavior+and+aging+specified+by+mitochondrial+function+during+development.">Rates of behavior and aging specified by mitochondrial function during development.</a> Science. 298, 2398-2401 (2002).</li><li>Hamilton, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+RNAi+screen+for+longevity+genes+in+C.+elegans.">A systematic RNAi screen for longevity genes in C. elegans.</a> Genes Dev. 19, 1544-1555 (2005).</li><li>Hansen, M., Hsu, A. L., Dillin, A., Kenyon, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+genes+tied+to+endocrine,+metabolic,+and+dietary+regulation+of+lifespan+from+a+Caenorhabditis+elegans+genomic+RNAi+screen.">New genes tied to endocrine, metabolic, and dietary regulation of lifespan from a Caenorhabditis elegans genomic RNAi screen.</a> PLoS Genet. 1, 119-128 (2005).</li><li>Lee, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+RNAi+screen+identifies+a+critical+role+for+mitochondria+in+C.+elegans+longevity.">A systematic RNAi screen identifies a critical role for mitochondria in C. elegans longevity.</a> Nat Genet. 33, 40-48 (2003).</li><li>Smith, E. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age-+and+calorie-independent+life+span+extension+from+dietary+restriction+by+bacterial+deprivation+in+Caenorhabditis+elegans.">Age- and calorie-independent life span extension from dietary restriction by bacterial deprivation in Caenorhabditis elegans.</a> BMC Dev Biol. 8, 49-49 (2008).</li><li>Hodgkin, J., Horvitz, H. R., Brenner, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nondisjunction+Mutants+of+the+Nematode+CAENORHABDITIS+ELEGANS.">Nondisjunction Mutants of the Nematode CAENORHABDITIS ELEGANS.</a> Genetics. 91, 67-94 (1979).</li><li>Emmons, S. W., Sternberg, P. W., Riddle, D. L., Blumenthal, T., Meyer, B. J., Priess, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> C. ELEGANS II. , (1997).</li><li>Lipton, J., Kleemann, G., Ghosh, R., Lints, R., Emmons, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mate+searching+in+Caenorhabditis+elegans:+a+genetic+model+for+sex+drive+in+a+simple+invertebrate.">Mate searching in Caenorhabditis elegans: a genetic model for sex drive in a simple invertebrate.</a> J Neurosci. 24, 7427-7434 (2004).</li><li>Davis, B. O., Anderson, G. L., Dusenbery, 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=Total+luminescence+spectroscopy+of+fluorescence+changes+during+aging+in+Caenorhabditis+elegans.">Total luminescence spectroscopy of fluorescence changes during aging in Caenorhabditis elegans.</a> Biochemistry. 21, 4089-4095 (1982).</li><li>Klass, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aging+in+the+nematode+Caenorhabditis+elegans:+major+biological+and+environmental+factors+influencing+life+span.">Aging in the nematode Caenorhabditis elegans: major biological and environmental factors influencing life span.</a> Mech Ageing Dev. 6, 413-429 (1977).</li><li>Steinkraus, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dietary+restriction+suppresses+proteotoxicity+and+enhances+longevity+by+an+hsf-1-dependent+mechanism+in+Caenorhabditis+elegans.">Dietary restriction suppresses proteotoxicity and enhances longevity by an hsf-1-dependent mechanism in Caenorhabditis elegans.</a> Aging Cell. 7, 394-404 (2008).</li></ol>

장수 유전자 제어 C. 광범위하게 연구되어 수명을 확인할 수있는 쉽고 인텔리로 주로 인해 엘레간스. 본 문서에서 언급한 다른 프로토콜은 C로 재현성 수명 데이터를 얻기위한 기본 프레임 워크를 설명 엘레간스하며 relatednematode 수종에도 적용할 수 있습니다. 2 이러한 절차는 다양한 조건 하에서 수명을 측정하기 위해 적응 수있는 몇 가지 간단한 변경을함으로?...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
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		<tbody>
		
<tr>
<td>Agar</td>
<td>Reagent</td>
<td>Fisher Scientific (BD Diagnostic Systems)</td>
<td>DF0145-17-0 (214530)</td>
<td>&nbsp;</td>
<tr>
<td>Ampicillin</td>
<td>Reagent</td>
<td>MidSci</td>
<td>0339</td>
<td>&nbsp;</td>
<tr>
<td>BactoTryptone</td>
<td>Reagent</td>
<td>Fisher Scientific (BD Diagnostic Systems)</td>
<td>DF0123-17-3 (211705)</td>
<td>&nbsp;</td>
<tr>
<td>BactoPeptone</td>
<td>Reagent</td>
<td>Fisher Scientific (BD Diagnostic Systems)</td>
<td>DF0118-17-0 (211677)</td>
<td>&nbsp;</td>
<tr>
<td>CaCl2</td>
<td>Reagent</td>
<td>Fisher Scientific (JT Baker)</td>
<td>NC9699248 (1332-01)</td>
<td>&nbsp;</td>
<tr>
<td>Carbenicillin</td>
<td>Reagent</td>
<td>Gold BioTechnology (GBT)</td>
<td>C0109</td>
<td>&nbsp;</td>
<tr>
<td>Cholesterol</td>
<td>Reagent</td>
<td>Sigma-Aldrich</td>
<td>C75209</td>
<td>&nbsp;</td>
<tr>
<td>Fluorodeoxyuridine (FUDR)</td>
<td>Reagent</td>
<td>Sigma-Aldrich</td>
<td>F0503</td>
<td>&nbsp;</td>
<tr>
<td>Isopropyl &#x03B2;-d-Thiogalactopyranoside (IPTG)</td>
<td>Reagent</td>
<td>Gold BioTechnology (GBT)</td>
<td>I2481C5</td>
<td>&nbsp;</td>
<tr>
<td>KPi Dibasic</td>
<td>Reagent</td>
<td>Fisher Scientific (JT Baker)</td>
<td>5087862 (3252-01)</td>
<td>&nbsp;</td>
<tr>
<td>KPi Monobasic</td>
<td>Reagent</td>
<td>Fisher Scientific (JT Baker)</td>
<td>5087861 (3246-01)</td>
<td>&nbsp;</td>
<tr>
<td>MgSO4</td>
<td>Reagent</td>
<td>Fisher Scientific (JT Baker)</td>
<td>NC9561800 (2500-01)</td>
<td>&nbsp;</td>
<tr>
<td>NaCl</td>
<td>Reagent</td>
<td>Fisher Scientific</td>
<td>S251</td>
<td>&nbsp;</td>
<tr>
<td>Tris</td>
<td>Reagent</td>
<td>Sigma-Aldrich</td>
<td>T1503</td>
<td>&nbsp;</td>
<tr>
<td>Yeast Extract</td>
<td>Reagent</td>
<td>Fisher Scientific (BD Diagnostic Systems)</td>
<td>DF0886-17-0 (288620)</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

measuring caenorhabditis elegans life span on solid media

노화는 세포의 구성 요소와 사망률의 결과 organelles의 점진적 저하 특징 퇴행성 과정이다. 선충류<em&gt; Caenorhabditis 엘레간스</em&gt; 노화의 생물학을 연구하는 데 사용하는 주요 모델로 등장했습니다. 거의 모든 생물 학적 서브 시스템이 점점 나이가 기능 감소를 겪습 때문에 노화의 총 비율을 고려할 때, 수명은 관심의 기본 종점이다. nematodes에서, 수명은 일반적으로 동물은 외부 자극에 반응 남아있는 일수로 정의됩니다. Nematodes은 액체 매체 또는 접시에있는 고체 매체에 중 전파 될 수 있으며, 기술은 두 조건에서 수명을 측정하기 위해 개발되었습니다. 여기 고체 선충류의 성장 미디어에 유지 nematodes의 수명을 측정하는 일반적인 프로토콜을 제시하고 UV - 살해 박테리아의 음식을 먹이. 이러한 절차는 쉽게 사는 박테리아,식이 제한, 그리고 RNA 간섭 구성된 식단을 포함하여 다양한 일반적인 조건 하에서 분석 수명을 적용할 수 있습니다.

Watch this Scientific Journal Video about Measuring Caenorhabditis elegans Life Span on Solid Media at JoVE.com

Measuring Caenorhabditis elegans Life Span on Solid Media

이 문서에서 우리는 UV - 살해 박테리아 음식 고체 미디어에 유지 nematodes의 수명을 측정하는 일반적인 프로토콜을 제시한다.

측정 Caenorhabditis 엘레간스</em솔리드 미디어> 수명

Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The nematode ...

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Measuring Caenorhabditis elegans Life Span on Solid Media

29.7K 조회수.  University of Washington. 이 문서에서 우리는 UV - 살해 박테리아 음식 고체 미디어에 유지 nematodes의 수명을 측정하는 일반적인 프로토콜을 제시한다.

비디오: Measuring Caenorhabditis elegans Life Span on Solid Media

Quantifying Yeast Chronological Life Span by Outgrowth of Aged Cells

The budding yeast Saccharomyces cerevisiae has proven to be an important model organism in the field of aging research 1. The replicative and chronological life spans are two established paradigms used to study aging in yeast. Replicative aging is defined as the number of daughter cells a single yeast mother cell produces before senescence; chronological aging is defined by the length of time cells can survive in a non-dividing, quiescence-like state 2. We have developed a high-throughput method for quantitative measurement of chronological life span. This method involves aging the cells in a defined medium under agitation and at constant temperature. At each age-point, a sub-population of cells is removed from the aging culture and inoculated into rich growth medium. A high-resolution growth curve is then obtained for this sub-population of aged cells using a Bioscreen C MBR machine. An algorithm is then applied to determine the relative proportion of viable cells in each sub-population based on the growth kinetics at each age-point. This method requires substantially less time and resources compared to other chronological lifespan assays while maintaining reproducibility and precision. The high-throughput nature of this assay should allow for large-scale genetic and chemical screens to identify novel longevity modifiers for further testing in more complex organisms.

Chronological aging in yeast refers to the loss of cell viability associated with time in stationary phase. Here we describe a high-throughput method for quantitatively determining yeast chronological life span.

지드 셀의 파생물로 효모 연대기의 수명 Quantifying

The budding yeast Saccharomyces cerevisiae has proven to be an important model organism in the field of aging research 1. The replicative and ...

연구

생물학

Measuring Replicative Life Span in the Budding Yeast

Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The budding yeast Saccharomyces cerevisiae has been used extensively to study the biology of aging, and several determinants of yeast longevity have been shown to be conserved in multicellular eukaryotes, including worms, flies, and mice 1. Due to the lack of easily quantified age-associated phenotypes, aging in yeast has been assayed almost exclusively by measuring the life span of cells in different contexts, with two different life span paradigms in common usage 2. Chronological life span refers to the length of time that a mother cell can survive in a non-dividing, quiescence-like state, and is proposed to serve as a model for aging of post-mitotic cells in multicellular eukaryotes. Replicative life span, in contrast, refers the number of daughter cells produced by a mother cell prior to senescence, and is thought to provide a model of aging in mitotically active cells. Here we present a generalized protocol for measuring the replicative life span of budding yeast mother cells. The goal of the replicative life span assay is to determine how many times each mother cell buds. The mother and daughter cells can be easily differentiated by an experienced researcher using a standard light microscope (total magnification 160X), such as the Zeiss Axioscope 40 or another comparable model. Physical separation of daughter cells from mother cells is achieved using a manual micromanipulator equipped with a fiber-optic needle. Typical laboratory yeast strains produce 20-30 daughter cells per mother and one life span experiment requires 2-3 weeks.

In this article we present a general protocol for measuring the replicative life span of yeast mother cells.

버딩 효모에 Replicative 수명 측정

Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The budding ...

Measuring Caenorhabditis elegans Life Span in 96 Well Microtiter Plates

Lifespan is a biological process regulated by several genetic pathways. One strategy to investigate the biology of aging is to study animals that harbor mutations in components of age-regulatory pathways. If these mutations perturb the function of the age-regulatory pathway and therefore alter the lifespan of the entire organism, they provide important mechanistic insights1-3.
Another strategy to investigate the regulation of lifespan is to use small molecules to perturb age-regulatory pathways. To date, a number of molecules are known to extend lifespan in various model organisms and are used as tools to study the biology of aging4-16. The number of molecules identified thus far is small compared to the genetic "toolset" that is available to study the biology of aging.
Caenorhabditis elegans is one of the principle models used to study aging because of its excellent genetics and short lifespan of three weeks. More recently, C.elegans has emerged as a model organism for phenotype based drug screens5,7,16-20 because of its small size and its ability to grow in microtiter plates.
Here we present an assay to measure C.elegans lifespan in 96 well microtiter plates. The assay was developed and successfully used to screen large libraries for molecules that extend C.elegans lifespan7. The reliability of the assay was evaluated in multiple tests: first, by measuring the lifespan of wild type animals grown at different temperatures; second, by measuring the lifespan of mutants with altered lifespans; third, by measuring changes in lifespan in response to different concentrations of the antidepressant Mirtazepine. Mirtazepine has previously been shown to extend lifespan in C.elegans7. The results of these tests show that the assay is able to replicate previous findings from other assays and is quantitative. The microtiter format also makes this lifespan assay compatible with automated liquid handling systems and allows integration into automated platforms.

Measuring Caenorhabditis elegans Life Span in 96 Well Microtiter Plates

In this protocol we present a method to measure Caenorhabditis elegans lifespan in 96 well microtiter plates.

측정 Caenorhabditis 엘레간스</em96 웰 플레이트에 Microtiter> 수명

Lifespan is a biological process regulated by several genetic pathways. One strategy to investigate the biology of aging is to study animals that harbor ...

Solid Plate-based Dietary Restriction in Caenorhabditis elegans

Reduction of food intake without malnutrition or starvation is known to increase lifespan and delay the onset of various age-related diseases in a wide range of species, including mammals. It also causes a decrease in body weight and fertility, as well as lower levels of plasma glucose, insulin, and IGF-1 in these animals. This treatment is often referred to as dietary restriction (DR) or caloric restriction (CR). The nematode Caenorhabditis elegans has emerged as an important model organism for studying the biology of aging. Both environmental and genetic manipulations have been used to model DR and have shown to extend lifespan in C. elegans. However, many of the reported DR studies in C. elegans were done by propagating animals in liquid media, while most of the genetic studies in the aging field were done on the standard solid agar in petri plates. Here we present a DR protocol using standard solid NGM agar-based plate with killed bacteria.

Solid Plate-based Dietary Restriction in Caenorhabditis elegans

Here we present a protocol for performing solid plate-based dietary restriction in C. elegans with killed bacteria.

솔리드 플레이트 기반식이 제한에 Caenorhabditis 엘레간스</em

Reduction of food intake without malnutrition or starvation is known to increase lifespan and delay the onset of various age-related diseases in a wide ...

Basic Caenorhabditis elegans Methods: Synchronization and Observation

Research into the molecular and developmental biology of the nematode Caenorhabditis elegans was begun in the early seventies by Sydney Brenner and it has since been used extensively as a model organism 1. C. elegans possesses key attributes such as simplicity, transparency and short life cycle that have made it a suitable experimental system for fundamental biological studies for many years 2. Discoveries in this nematode have broad implications because many cellular and molecular processes that control animal development are evolutionary conserved 3.
C. elegans life cycle goes through an embryonic stage and four larval stages before animals reach adulthood. Development can take 2 to 4 days depending on the temperature. In each of the stages several characteristic traits can be observed. The knowledge of its complete cell lineage 4,5 together with the deep annotation of its genome turn this nematode into a great model in fields as diverse as the neurobiology 6, aging 7,8, stem cell biology 9 and germ line biology 10. 
An additional feature that makes C. elegans an attractive model to work with is the possibility of obtaining populations of worms synchronized at a specific stage through a relatively easy protocol. The ease of maintaining and propagating this nematode added to the possibility of synchronization provide a powerful tool to obtain large amounts of worms, which can be used for a wide variety of small or high-throughput experiments such as RNAi screens, microarrays, massive sequencing, immunoblot or in situ hybridization, among others. 
Because of its transparency, C. elegans structures can be distinguished under the microscope using Differential Interference Contrast microscopy, also known as Nomarski microscopy. The use of a fluorescent DNA binder, DAPI (4',6-diamidino-2-phenylindole), for instance, can lead to the specific identification and localization of individual cells, as well as subcellular structures/defects associated to them.

Basic Caenorhabditis elegans Methods: Synchronization and Observation

The easiness of maintaining and propagating the nematode C. elegans make it a nice model organism to work with. The possibility of synchronizing worms allows the work with a significant amount of subjects at the same developmental stage, what facilitates the study of one particular process in many animals.

기본<em&gt; Caenorhabditis elegans</em&gt; 방법 : 동기화 및 전망

Research into the molecular and developmental biology of the nematode Caenorhabditis elegans was begun in the early seventies by Sydney Brenner and it has ...

Culturing Caenorhabditis elegans in Axenic Liquid Media and Creation of Transgenic Worms by Microparticle Bombardment

In this protocol, we present the required materials, and the procedure for making modified&#160;C. elegans&#160;Habituation and Reproduction media (mCeHR). Additionally, the steps for exposing and acclimatizing C. elegans grown on E. coli to axenic liquid media are described. Finally, downstream experiments that utilize axenic C. elegans illustrate the benefits of this procedure. The ability to analyze and determine C. elegans&#160;nutrient requirement was illustrated by growing N2 wild type worms in axenic liquid media with varying heme concentrations. This procedure can be replicated with other nutrients to determine the optimal concentration for worm growth and development or, to determine the toxicological effects of drug treatments. The effects of varied heme concentrations on the growth of wild type worms were determined through qualitative microscopic observation and by quantitating the number of worms that grew in each heme concentration. In addition, the effect of varied nutrient concentrations can be assayed by utilizing worms that express fluorescent sensors that respond to changes in the nutrient of interest. Furthermore, a large number of worms were easily produced for the generation of transgenic C. elegans using microparticle bombardment.

Culturing Caenorhabditis elegans in Axenic Liquid Media and Creation of Transgenic Worms by Microparticle Bombardment

C. elegans is usually grown on solid agar plates or in liquid cultures seeded with E. coli. To prevent bacterial byproducts from confounding toxicological and nutritional studies, we utilized an axenic liquid medium, CeHR, to grow and synchronize a large number of worms for a range of downstream applications.

배양<em&gt; 예쁜 꼬마 선충</em &gt; 미세 폭격에 의한 무균 액체 미디어 및 형질 전환 웜의 생성에

In this protocol, we present the required materials, and the procedure for making modified&#160;C. elegans&#160;Habituation and Reproduction media ...

Measuring Oxidative Stress Resistance of Caenorhabditis elegans in 96-well Microtiter Plates

Oxidative stress, which is the result of an imbalance between production and detoxification of reactive oxygen species, is a major contributor to chronic human disorders, including cardiovascular and neurodegenerative diseases, diabetes, aging, and cancer. Therefore, it is important to study oxidative stress not only in cell systems but also using whole organisms. C. elegans is an attractive model organism to study the genetics of oxidative stress signal transduction pathways, which are highly evolutionarily conserved.
Here, we provide a protocol to measure oxidative stress resistance in C. elegans in liquid. Briefly, ROS-inducing reagents such as paraquat (PQ) and H2O2 are dissolved in M9 buffer, and solutions are aliquoted in the wells of a 96 well microtiter plate. Synchronized L4/young adult C. elegans animals are transferred to the wells (5-8 animals/well) and survival is measured every hour until most worms are dead. When performing an oxidative stress resistance assay using a low concentration of stressors in plates, aging might influence the behavior of animals upon oxidative stress, which could lead to an incorrect interpretation of the data. However, in the assay described herein, this problem is unlikely to occur since only L4/young adult animals are being used. Moreover, this protocol is inexpensive and results are obtained in one day, which renders this technique attractive for genetic screens. Overall, this will help to understand oxidative stress signal transduction pathways, which could be translated into better characterization of oxidative stress-associated human disorders.

Measuring Oxidative Stress Resistance of Caenorhabditis elegans in 96-well Microtiter Plates

C. elegans is an attractive model organism to study signal transduction pathways involved in oxidative stress resistance. Here we provide a protocol to measure oxidative stress resistance of C. elegans animals in liquid phase, using several oxidizing agents in 96 well plates.

의 산화 스트레스 저항을 측정<em&gt; 예쁜 꼬마 선충</em&gt; 96 웰 마이크로 타이 터 플레이트에

Oxidative stress, which is the result of an imbalance between production and detoxification of reactive oxygen species, is a major contributor to chronic ...

Using Caenorhabditis elegans to Screen for Tissue-Specific Chaperone Interactions

Correct folding and assembly of proteins and protein complexes are essential for cellular function. Cells employ quality control pathways that correct, sequester or eliminate damaged proteins to maintain a healthy proteome, thus ensuring cellular proteostasis and preventing further protein damage. Because of redundant functions within the proteostasis network, screening for detectable phenotypes using knockdown or mutations in chaperone-encoding genes in the multicellular organism Caenorhabditis elegans results in the detection of minor or no phenotypes in most cases. We have developed a targeted screening strategy to identify chaperones required for a specific function and thus bridge the gap between phenotype and function. Specifically, we monitor novel chaperone interactions using RNAi synthetic interaction screens, knocking-down chaperone expression, one chaperone at a time, in animals carrying a mutation in a chaperone-encoding gene or over-expressing a chaperone of interest. By disrupting two chaperones that individually present no gross phenotype, we can identify chaperones that aggravate or expose a specific phenotype when both perturbed. We demonstrate that this approach can identify specific sets of chaperones that function together to modulate the folding of a protein or protein complexes associated with a given phenotype.

Using Caenorhabditis elegans to Screen for Tissue-Specific Chaperone Interactions

To study chaperone-chaperone and chaperone-substrate interactions, we perform synthetic interaction screens in Caenorhabditis elegans using RNA interference in combination with mild mutations or over-expression of chaperones and monitor tissue-specific protein dysfunction at the organismal level.

조직별 샤페론 상호 작용을 위해 검사하기 위해 Caenorhabditis elegans를 사용

Correct folding and assembly of proteins and protein complexes are essential for cellular function. Cells employ quality control pathways that correct, ...

Label-Free Imaging of Lipid Storage Dynamics in Caenorhabditis elegans using Stimulated Raman Scattering Microscopy

Lipid metabolism is a fundamental physiological process necessary for cellular and organism health. Dysregulation of lipid metabolism often elicits obesity and many associated diseases including cardiovascular disorders, type II diabetes, and cancer. To advance the current understanding of lipid metabolic regulation, quantitative methods to precisely measure in vivo lipid storage levels in time and space have become increasingly important and useful. Traditional approaches to analyze lipid storage are semi-quantitative for microscopic assessment or lacking spatio-temporal information for biochemical measurement. Stimulated Raman scattering (SRS) microscopy is a label-free chemical imaging technology that enables rapid and quantitative detection of lipids in live cells with a subcellular resolution. As the contrast is exploited from intrinsic molecular vibrations, SRS microscopy also permits four-dimensional tracking of lipids in live animals. In the last decade, SRS microscopy has been widely used for small molecule imaging in biomedical research and overcome the major limitations of conventional fluorescent staining and lipid extraction methods. In the laboratory, we have combined SRS microscopy with the genetic and biochemical tools available to the powerful model organism, Caenorhabditis elegans, to investigate the distribution and heterogeneity of lipid droplets across different cells and tissues and ultimately to discover novel conserved signaling pathways that modulate lipid metabolism. Here, we present the working principles and the detailed setup of the SRS microscope and provide methods for its use in quantifying lipid storage at distinct developmental timepoints of wild-type and insulin signaling deficient mutant C. elegans.

Label-Free Imaging of Lipid Storage Dynamics in Caenorhabditis elegans using Stimulated Raman Scattering Microscopy

Stimulated Raman scattering (SRS) microscopy allows selective, label-free imaging of specific chemical moieties and it has been effectively employed to image lipid molecules in vivo. Here, we provide a brief introduction to the principle of SRS microscopy and describe methods for its use in imaging lipid storage in Caenorhabditis elegans.

자극라만 산란 현미경 검사를 사용하여 Caenorhabditis elegans의 지질 저장 역학의 라벨 프리 이미징

Lipid metabolism is a fundamental physiological process necessary for cellular and organism health. Dysregulation of lipid metabolism often elicits ...

Automating Aggregate Quantification in Caenorhabditis elegans

A rise in the prevalence of neurodegenerative protein conformational diseases (PCDs) has fostered a great interest in this subject over the years. This increased attention has called for the diversification and improvement of animal models capable of reproducing disease phenotypes observed in humans with PCDs. Though murine models have proven invaluable, they are expensive and are associated with laborious, low-throughput methods. Use of the Caenorhabditis elegans nematode model to study PCDs has been justified by its relative ease of maintenance, low cost, and rapid generation time, which allow for high-throughput applications. Additionally, high conservation between the C. elegans and human genomes makes this model an invaluable discovery tool. Nematodes that express fluorescently tagged tissue-specific polyglutamine (polyQ) tracts exhibit age- and polyQ length-dependent aggregation characterized by fluorescent foci. Such reporters are often employed as proxies to monitor changes in proteostasis across tissues. Manual aggregate quantification is time-consuming, limiting experimental throughput. Furthermore, manual foci quantification can introduce bias, as aggregate identification can be highly subjective. Herein, a protocol consisting of worm culturing, image acquisition, and data processing was standardized to support high-throughput aggregate quantification using C. elegans that express intestine-specific polyQ. By implementing a C. elegans-based image processing pipeline using CellProfiler, an image analysis software, this method has been optimized to separate and identify individual worms and enumerate their respective aggregates. Though the concept of automation is not entirely unique, the need to standardize such procedures for reproducibility, elimination of bias from manual counting, and increase throughput is high. It is anticipated that these methods can drastically simplify the screening process of large bacterial, genomic, or drug libraries using the C. elegans model.

Automating Aggregate Quantification in Caenorhabditis elegans

The following protocol describes the development and optimization of a high-throughput workflow for worm culturing, fluorescence imaging, and automated image processing to quantify polyglutamine aggregates as an assessment of changes in proteostasis.

Caenorhabditis elegans의 집계 정량화 자동화

A rise in the prevalence of neurodegenerative protein conformational diseases (PCDs) has fostered a great interest in this subject over the years. This ...

측정 Caenorhabditis 엘레간스 수명