graticulesoptics EM GRIDS产品列表,欢迎订购


Graticules Optics 在提供玻璃、薄膜和金属箔上的光刻产品方面是仅有的,此外还提供涂层、粘合、安装和小型光学组件。

graticulesoptics EM GRIDS产品列表,欢迎订购!

货号 Mesh Size Material 价格 品牌
09D00901 50 MESH Copper 120Vial 234.1171571 graticulesoptics
09D01901 50 MESH Nickel 120Vial 304.6883576 graticulesoptics
09D00903 100 MESH Copper 120Vial 234.1171571 graticulesoptics
09D01903 100 MESH Nickel 120Vial 304.6883576 graticulesoptics
09D00904 150 MESH Copper 120Vial 257.6408906 graticulesoptics
09D01904 150 MESH Nickel 120Vial 327.8386985 graticulesoptics
09D00905 200 MESH Copper 120Vial 257.6408906 graticulesoptics
09D01905 200 MESH Nickel 120Vial 327.8386985 graticulesoptics
09D00906 300 MESH Copper 120Vial 257.6408906 graticulesoptics
09D01906 300 MESH Nickel 120Vial 327.8386985 graticulesoptics
09D00907 400 MESH Copper 120Vial 281.1646241 graticulesoptics
09D01907 400 MESH Nickel 120Vial 339.7872616 graticulesoptics
07D00901 50 MESH Copper 100Vial 369.3664 graticulesoptics
07D01901 50 MESH Nickel 100Vial 416.763962 graticulesoptics
07D02901 50 MESH Gold 25Vial 614.1515482 graticulesoptics
08L07901 50 MESH Titanium 25Vial 2102.1 graticulesoptics
08L05901 50 MESH Molybdenum 25Vial 1103.823608 graticulesoptics
08L04901 50 MESH Aluminium 25Vial 925.8935111 graticulesoptics
08L06901 50 MESH Stainless Steel 25Vial 772.4647971 graticulesoptics
07D00902 75 MESH Copper 100Vial 369.6368 graticulesoptics
03D00650 75 MESH Copper 100Vial 369.6426155 graticulesoptics
03D00700 75 MESH Copper with Rhodium 100Vial 430.04 graticulesoptics
07D01902 75 MESH Nickel 100Vial 416.7610912 graticulesoptics
03D00651 75 MESH Nickel 100Vial 416.7610912 graticulesoptics
03D00701 75 MESH Nickel ( High Grade ) 100Vial 449.4048 graticulesoptics
07D02902 75 MESH Gold 25Vial 614.1515482 graticulesoptics
03D00702 75 MESH Gold ( High Grade ) 100Vial 3188.886004 graticulesoptics
08L07902 75 MESH Titanium 25Vial 2103.4 graticulesoptics
08L05902 75 MESH Molybdenum 25Vial 1111.102181 graticulesoptics
08L04902 75 MESH Aluminium 25Vial 925.9330304 graticulesoptics
08L06902 75 MESH Stainless Steel 25Vial 777.6421415 graticulesoptics
07D00903 100 MESH Copper 100Vial 369.6426155 graticulesoptics
03D00652 100 MESH Copper 100Vial 369.6426155 graticulesoptics
03D00703 100 MESH Copper with Rhodium 100Vial 430.04 graticulesoptics
07D01903 100 MESH Nickel 100Vial 416.7610912 graticulesoptics
03D00653 100 MESH Nickel 100Vial 416.7610912 graticulesoptics
03D00704 100 MESH Nickel ( High Grade ) 100Vial 449.44312 graticulesoptics
07D02903 100 MESH Gold 25Vial 614.1515482 graticulesoptics
03D00705 100 MESH Gold ( High Grade ) 100Vial 3188.886004 graticulesoptics
08L07903 100 MESH Titanium 25Vial 2103.4 graticulesoptics
08L05903 100 MESH Molybdenum 25Vial 1111.102181 graticulesoptics
08L04903 100 MESH Aluminium 25Vial 925.9330304 graticulesoptics
08L06903 100 MESH Stainless Steel 25Vial 777.6421415 graticulesoptics
07D00904 150 MESH Copper 100Vial 369.6426155 graticulesoptics
03D00654 150 MESH Copper 100Vial 369.6426155 graticulesoptics
03D00706 150 MESH Copper with Rhodium 100Vial 430.04 graticulesoptics
07D01904 150 MESH Nickel 100Vial 416.7610912 graticulesoptics
03D00655 150 MESH Nickel 100Vial 416.7610912 graticulesoptics
03D00707 150 MESH Nickel ( High Grade ) 100Vial 449.44312 graticulesoptics
07D02904 150 MESH Gold 25Vial 614.1515482 graticulesoptics
03D00708 150 MESH Gold ( High Grade ) 100Vial 3188.886004 graticulesoptics
08L07904 150 MESH Titanium 25Vial 2103.4 graticulesoptics
08L05904 150 MESH Molybdenum 25Vial 1111.102181 graticulesoptics
08L04904 150 MESH Aluminium 25Vial 925.9330304 graticulesoptics
08L06904 150 MESH Stainless Steel 25Vial 777.6421415 graticulesoptics
07D00905 200 MESH Copper 100Vial 369.6426155 graticulesoptics
03D00656 200 MESH Copper 100Vial 369.6426155 graticulesoptics
03D00709 200 MESH Copper with Rhodium 100Vial 430.04 graticulesoptics
07D01905 200 MESH Nickel 100Vial 416.7610912 graticulesoptics
03D00657 200 MESH Nickel 100Vial 414.1917931 graticulesoptics
03D00710 200 MESH Nickel ( High Grade ) 100Vial 449.44312 graticulesoptics
07D02905 200 MESH Gold 25Vial 614.1515482 graticulesoptics
03D00711 200 MESH Gold ( High Grade ) 100Vial 3188.886004 graticulesoptics
08L07905 200 MESH Titanium 25Vial 2103.4 graticulesoptics
08L05905 200 MESH Molybdenum 25Vial 1111.102181 graticulesoptics
08L04905 200 MESH Aluminium 25Vial 925.9330304 graticulesoptics
08L06905 200 MESH Stainless Steel 25Vial 777.6421415 graticulesoptics
07D00906 300 MESH Copper 100Vial 369.6426155 graticulesoptics
03D00658 300 MESH Copper 100Vial 369.6426155 graticulesoptics
03D00712 300 MESH Copper with Rhodium 100Vial 430.04 graticulesoptics
07D01906 300 MESH Nickel 100Vial 416.3958317 graticulesoptics
03D00659 300 MESH Nickel 100Vial 416.7610912 graticulesoptics
03D00713 300 MESH Nickel ( High Grade ) 100Vial 449.44312 graticulesoptics
07D02906 300 MESH Gold 25Vial 614.1515482 graticulesoptics
03D00714 300 MESH Gold ( High Grade ) 100Vial 3188.886004 graticulesoptics
08L07906 300 MESH Titanium 25Vial 2808 graticulesoptics
08L05906 300 MESH Molybdenum 25Vial 1111.102181 graticulesoptics
08L06906 300 MESH Stainless Steel 25Vial 777.4730034 graticulesoptics
08L04906 300 MESH Aluminium 25Vial 925.990593 graticulesoptics
07D00907 400 MESH Copper 100Vial 382.9832623 graticulesoptics
03D00661 400 MESH Copper 100Vial 382.9832623 graticulesoptics
03D00715 400 MESH Copper with Rhodium 100Vial 430.04 graticulesoptics
07D01907 400 MESH Nickel 100Vial 462.0359196 graticulesoptics
03D00662 400 MESH Nickel 100Vial 462.0359196 graticulesoptics
03D00716 400 MESH Nickel ( High Grade ) 100Vial 449.44312 graticulesoptics
07D02907 400 MESH Gold 25Vial 670.8777724 graticulesoptics
03D00717 400 MESH Gold ( High Grade ) 100Vial 3260.147913 graticulesoptics
08L07907 400 MESH Titanium 25Vial 3146 graticulesoptics
08L04907 400 MESH Aluminium 25Vial 978.1806969 graticulesoptics
07D00908 500 MESH Copper 100Vial 485.0399432 graticulesoptics
07D01908 500 MESH Nickel 100Vial 555.1300433 graticulesoptics
07D02908 500 MESH Gold 25Vial 792.1997547 graticulesoptics
07D00960 600 MESH Copper 100Vial 555.1944423 graticulesoptics
07D01961 600 MESH Nickel 100Vial 555.4536152 graticulesoptics
07D00961 600 MESH Copper with Rhodium 100Vial 544.7 graticulesoptics
07D30901 50 MESH Copper 100Vial 499.3875109 graticulesoptics
07D31901 50 MESH Nickel 100Vial 555.1300433 graticulesoptics
07D32901 50 MESH Gold 25Vial 784.6581179 graticulesoptics
08L37901 50 MESH Titanium 25Vial 2103.4 graticulesoptics
08L35901 50 MESH Molybdenum 25Vial 1111.102181 graticulesoptics
08L36901 50 MESH Stainless Steel 25Vial 777.6421415 graticulesoptics
08L34901 50 MESH Aluminium 25Vial 925.990593 graticulesoptics
07D30902 75 MESH Copper 100Vial 499.3875109 graticulesoptics
07D31902 75 MESH Nickel 100Vial 555.1300433 graticulesoptics
07D32902 75 MESH Gold 25Vial 784.6581179 graticulesoptics
08L37902 75 MESH Titanium 25Vial 2103.4 graticulesoptics
08L35902 75 MESH Molybdenum 25Vial 1111.102181 graticulesoptics
08L34902 75 MESH Aluminium 25Vial 1226.576193 graticulesoptics
08L36902 75 MESH Stainless Steel 25Vial 1017.856478 graticulesoptics
07D30903 100 MESH Copper 100Vial 499.3875109 graticulesoptics
07D31903 100 MESH Nickel 100Vial 555.1300433 graticulesoptics
07D30904 150 MESH Copper 100Vial 499.3875109 graticulesoptics
07D31904 150 MESH Nickel 100Vial 555.1300433 graticulesoptics
07D32904 150 MESH Gold 25Vial 792.1997547 graticulesoptics
07D30905 200 MESH Copper 100Vial 499.3875109 graticulesoptics
07D31905 200 MESH Nickel 100Vial 555.1300433 graticulesoptics
07D32905 200 MESH Gold 25Vial 824.6615823 graticulesoptics
08L37905 200 MESH Titanium 25Vial 2103.4 graticulesoptics
08L35905 200 MESH Molybdenum 25Vial 1111.102181 graticulesoptics
08L34905 200 MESH Aluminium 25Vial 1226.576193 graticulesoptics
08L36905 200 MESH Stainless Steel 25Vial 1017.856478 graticulesoptics
07D30906 300 MESH Copper 100Vial 499.3875109 graticulesoptics
07D31906 300 MESH Nickel 100Vial 555.1300433 graticulesoptics
07D32906 300 MESH Gold 25Vial 824.6615823 graticulesoptics
08L35906 300 MESH Molybdenum 25Vial 1111.102181 graticulesoptics
08L37906 300 MESH Titanium 25Vial 2103.4 graticulesoptics
08L34906 300 MESH Aluminium 25Vial 1226.576193 graticulesoptics
08L36906 300 MESH Stainless Steel 25Vial 1017.856478 graticulesoptics
07D30907 400 MESH Copper 100Vial 532.5483532 graticulesoptics
07D31907 400 MESH Nickel 100Vial 601.3635555 graticulesoptics
07D32907 400 MESH Gold 25Vial 952.2136124 graticulesoptics
07D30908 500 MESH Copper 100Vial 601.3635555 graticulesoptics
07D31908 500 MESH Nickel 100Vial 740.0157501 graticulesoptics
07D32908 500 MESH OBSOLETE – Gold 25Vial 0 graticulesoptics
07D30960 600 MESH Copper 100Vial 601.4606459 graticulesoptics
07D31961 600 MESH Nickel 100Vial 740.0157501 graticulesoptics
07D30961 600 MESH Cu/Rh 100Vial 635.7927667 graticulesoptics
07D00909 254micron PITCH Copper 100Vial 401.0549327 graticulesoptics
07D01909 254micron PITCH Nickel 100Vial 440.5029588 graticulesoptics
03D00808 165micron PITCH Copper 100Vial 401.0549327 graticulesoptics
03D00809 165micron PITCH Nickel 100Vial 440.5029588 graticulesoptics
07D00910 127micron PITCH Copper 100Vial 401.0549327 graticulesoptics
03D00810 127micron PITCH Copper 100Vial 401.0549327 graticulesoptics
07D01910 127micron PITCH Nickel 100Vial 440.5029588 graticulesoptics
03D00811 127micron PITCH Nickel 100Vial 440.5029588 graticulesoptics
07D00911 84micron PITCH Copper 100Vial 401.0549327 graticulesoptics
07D01911 84micron PITCH Nickel 100Vial 440.5029588 graticulesoptics
07D30909 254micron PITCH Copper 100Vial 499.3875109 graticulesoptics
07D31909 254micron PITCH Nickel 100Vial 555.1300433 graticulesoptics
07D30910 127micron PITCH Copper 100Vial 499.3875109 graticulesoptics
07D31910 127micron PITCH Nickel 100Vial 555.1300433 graticulesoptics
07D30911 84micron PITCH Copper 100Vial 499.3875109 graticulesoptics
07D31911 84micron PITCH Nickel 100Vial 555.1300433 graticulesoptics
07D00912 254micron PITCH Copper 100Vial 401.0549327 graticulesoptics
07D01912 254micron PITCH Nickel 100Vial 468.9236267 graticulesoptics
07D02912 254micron PITCH Gold 25Vial 678.0915118 graticulesoptics
03D00804 165micron PITCH Copper 100Vial 401.0549327 graticulesoptics
03D00805 165micron PITCH Nickel 100Vial 440.5029588 graticulesoptics
07D00913 127micron PITCH Copper 100Vial 401.0549327 graticulesoptics
03D00807 127micron PITCH Copper 100Vial 401.0549327 graticulesoptics
07D01913 127micron PITCH Nickel 100Vial 440.5029588 graticulesoptics
03D00806 127micron PITCH Nickel 100Vial 440.7464652 graticulesoptics
07D02913 127micron PITCH Gold 25Vial 642.3507117 graticulesoptics
07D00914 84micron PITCH Copper 100Vial 401.0549327 graticulesoptics
07D01914 84micron PITCH Nickel 100Vial 440.5029588 graticulesoptics
07D02914 84micron PITCH Gold 25Vial 642.3507117 graticulesoptics
07D30912 254micron PITCH Copper 100Vial 499.3875109 graticulesoptics
07D31912 254micron PITCH Nickel 100Vial 555.1300433 graticulesoptics
07D30913 127micron PITCH Copper 100Vial 499.3875109 graticulesoptics
07D31913 127micron PITCH Nickel 100Vial 555.1300433 graticulesoptics
07D30914 84micron PITCH Copper 100Vial 499.3875109 graticulesoptics
07D31914 84micron PITCH Nickel 100Vial 555.1300433 graticulesoptics
07D00915 50/200 Copper 100Vial 499.3875109 graticulesoptics
07D01915 50/200 Nickel 100Vial 555.1300433 graticulesoptics
07D00915R 50/200 Copper with Rhodium 100Vial 613.08 graticulesoptics
07D00916 75/300 Copper 100Vial 499.3875109 graticulesoptics
03D00800 75/300 Copper 100Vial 499.3875109 graticulesoptics
03D00718 75/300 Copper with Rhodium 100Vial 613.34 graticulesoptics
07D01916 75/300 Nickel 100Vial 592.816171 graticulesoptics
03D00801 75/300 Nickel 100Vial 592.816171 graticulesoptics
03D00719 75/300 Nickel ( High Grade ) 100Vial 634.1118523 graticulesoptics
07D02916 75/300 Gold 25Vial 840.4006503 graticulesoptics
03D00720 75/300 Gold ( High Grade ) 100Vial 3096.005829 graticulesoptics
07D00970 75/300 STAGGERED PATTERN Copper 100Vial 500.0272452 graticulesoptics
07D00971 75/300 STAGGERED PATTERN Copper with Rhodium 100Vial 613.34 graticulesoptics
07D01970 75/300 STAGGERED PATTERN Nickel 100Vial 592.3768665 graticulesoptics
07D02970 75/300 STAGGERED PATTERN Gold 25Vial 840.4006503 graticulesoptics
07D00917 100/400 Copper 100Vial 384.3921421 graticulesoptics
03D00802 100/400 Copper 100Vial 384.3921421 graticulesoptics
07D01917 100/400 Nickel 100Vial 462.0359196 graticulesoptics
03D00803 100/400 Nickel 100Vial 462.0359196 graticulesoptics
03D00825 100/400 Gold 100Vial 2847.099027 graticulesoptics
07D30915 50/200 Copper 100Vial 499.3875109 graticulesoptics
07D31915 50/200 Nickel 100Vial 555.1300433 graticulesoptics
07D30916 75/300 Copper 100Vial 499.3875109 graticulesoptics
07D31916 75/300 Nickel 100Vial 555.1300433 graticulesoptics
07D00927 50/50 Copper 100Vial 694.1584771 graticulesoptics
07D01927 50/50 Nickel 100Vial 823.6779218 graticulesoptics
07D02927 50/50 Gold 25Vial 1131.245511 graticulesoptics
07D00928 100/100 Copper 100Vial 694.1584771 graticulesoptics
07D01928 100/100 Nickel 100Vial 823.6779218 graticulesoptics
07D02928 100/100 Gold 25Vial 1149.279859 graticulesoptics
08L07928 100/100 Titanium 25Vial 4186 graticulesoptics
08L05928 100/100 Molybdenum 25Vial 2188.051058 graticulesoptics
08L06928 100/100 Stainless Steel 25Vial 1235.188939 graticulesoptics
07D00929 100/200 Copper 100Vial 694.1584771 graticulesoptics
07D01929 100/200 Nickel 100Vial 823.6779218 graticulesoptics
07D02929 100/200 Gold 25Vial 1149.279859 graticulesoptics
08L07929 100/200 Titanium 25Vial 4186 graticulesoptics
08L05929 100/200 Molybdenum 25Vial 2188.051058 graticulesoptics
08L06929 100/200 Stainless Steel 25Vial 1235.188939 graticulesoptics
07D00930 100/300 Copper 100Vial 694.1584771 graticulesoptics
07D01930 100/300 Nickel 100Vial 823.6779218 graticulesoptics
07D02930 100/300 Gold 25Vial 1149.279859 graticulesoptics
08L07930 100/300 Titanium 25Vial 4758 graticulesoptics
08L06930 100/300 Stainless Steel 25Vial 1235.188939 graticulesoptics
07D00931 100/400 Copper 100Vial 694.1584771 graticulesoptics
07D01931 100/400 Nickel 100Vial 823.6779218 graticulesoptics
07D00948 90 MESH Copper 100Vial 370.4774944 graticulesoptics
07D01948 90 MESH Nickel 100Vial 416.7610912 graticulesoptics
07D02948 90 MESH Gold 25Vial 0 graticulesoptics
07D00949 135 MESH Copper 100Vial 370.4774944 graticulesoptics
07D01949 135 MESH Nickel 100Vial 416.7610912 graticulesoptics
07D02949 135 MESH Gold 25Vial 611.5283702 graticulesoptics
07D00950 180 MESH Copper 100Vial 370.4774944 graticulesoptics
07D01950 180 MESH Nickel 100Vial 416.7610912 graticulesoptics
07D02950 180 MESH Gold 25Vial 611.5283702 graticulesoptics
07D00951 270 MESH Copper 100Vial 370.4774944 graticulesoptics
07D01951 270 MESH Nickel 100Vial 416.7610912 graticulesoptics
07D02951 270 MESH Gold 25Vial 611.5283702 graticulesoptics
07D00952 360 MESH Copper 100Vial 370.4774944 graticulesoptics
07D01952 360 MESH Nickel 100Vial 416.7610912 graticulesoptics
07D02952 360 MESH Gold 25Vial 611.5283702 graticulesoptics
03D00603 H5 PATTERN Copper 100Vial 401.0549327 graticulesoptics
03D00604 H5 PATTERN Nickel 100Vial 440.5029588 graticulesoptics
03D00605 H5 PATTERN Gold 100Vial 3188.886004 graticulesoptics
07D30948 90 MESH Copper 100Vial 499.3875109 graticulesoptics
07D31948 90 MESH Nickel 100Vial 555.1300433 graticulesoptics
07D30949 135 MESH Copper 100Vial 499.3875109 graticulesoptics
07D30950 180 MESH Copper 100Vial 499.3875109 graticulesoptics
07D31950 180 MESH Nickel 100Vial 555.1300433 graticulesoptics
07D32950 180 MESH Gold 25Vial 791.5439602 graticulesoptics
07D30951 270 MESH Copper 100Vial 499.3875109 graticulesoptics
07D31951 270 MESH Nickel 100Vial 555.1300433 graticulesoptics
07D32951 270 MESH Gold 25Vial 791.5439602 graticulesoptics
07D30952 360 MESH Copper 100Vial 499.3875109 graticulesoptics
07D31952 360 MESH Nickel 100Vial 555.1300433 graticulesoptics
07D32952 360 MESH Gold 25Vial 791.5439602 graticulesoptics
07D00918 0.4mm HOLE GRIDS Copper 100Vial 532.5483532 graticulesoptics
07D01918 0.4mm HOLE GRIDS Nickel 100Vial 624.9590071 graticulesoptics
07D02918 0.4mm HOLE GRIDS Gold 25Vial 0 graticulesoptics
08L07918 0.4mm HOLE GRIDS Titanium 25Vial 3068 graticulesoptics
08L05918 0.4mm HOLE GRIDS Molybdenum 25Vial 1312.942613 graticulesoptics
08L06918 0.4mm HOLE GRIDS Stainless Steel 25Vial 1110.587984 graticulesoptics
07D00919 0.6mm HOLE GRIDS Copper 100Vial 532.5483532 graticulesoptics
03D00814 0.6mm HOLE GRIDS Copper 100Vial 532.5483532 graticulesoptics
07D01919 0.6mm HOLE GRIDS Nickel 100Vial 624.9590071 graticulesoptics
03D00815 0.6mm HOLE GRIDS Nickel 100Vial 624.9590071 graticulesoptics
07D02919 0.6mm HOLE GRIDS Gold 25Vial 878.7646285 graticulesoptics
08L05919 0.6mm HOLE GRIDS Molybdenum 25Vial 1312.942613 graticulesoptics
07D00920 0.8mm HOLE GRIDS Copper 100Vial 532.5483532 graticulesoptics
07D01920 0.8mm HOLE GRIDS Nickel 100Vial 624.9590071 graticulesoptics
07D00921 1.0mm HOLE GRIDS Copper 100Vial 532.5483532 graticulesoptics
03D00812 1.0mm HOLE GRIDS Copper 100Vial 532.5483532 graticulesoptics
07D01921 1.0mm HOLE GRIDS Nickel 100Vial 624.9590071 graticulesoptics
03D00813 1.0mm HOLE GRIDS Nickel 100Vial 624.9590071 graticulesoptics
07D02921 1.0mm HOLE GRIDS Gold 25Vial 878.7646285 graticulesoptics
08L07921 1.0mm HOLE GRIDS Titanium 25Vial 3068 graticulesoptics
08L05921 1.0mm HOLE GRIDS Molybdenum 25Vial 1312.942613 graticulesoptics
08L04921 1.0mm HOLE GRIDS Aluminium 25Vial 1303.391567 graticulesoptics
08L06921 1.0mm HOLE GRIDS Stainless Steel 25Vial 1110.587984 graticulesoptics
07D00922 1.5mm HOLE GRIDS Copper 100Vial 532.5483532 graticulesoptics
07D01922 1.5mm HOLE GRIDS Nickel 100Vial 624.9590071 graticulesoptics
08L07922 1.5mm HOLE GRIDS Titanium 25Vial 3068 graticulesoptics
08L05922 1.5mm HOLE GRIDS Molybdenum 25Vial 1312.942613 graticulesoptics
08L04922 1.5mm HOLE GRIDS Aluminium 25Vial 1341.839806 graticulesoptics
08L06922 1.5mm HOLE GRIDS Stainless Steel 25Vial 1110.587984 graticulesoptics
07D30980 1.5mm HOLE GRIDS WITH HANDLE Copper 100Vial 532.4176 graticulesoptics
07D31980 1.5mm HOLE GRIDS WITH HANDLE Nickel 100Vial 625.3000547 graticulesoptics
07D00923 2.0mm HOLE GRIDS Copper 100Vial 532.5483532 graticulesoptics
07D01923 2.0mm HOLE GRIDS Nickel 100Vial 624.9590071 graticulesoptics
07D00924 2mm x 0.980mm HOLE GRIDS Copper 100Vial 532.5483532 graticulesoptics
07D01924 2mm x 0.980mm HOLE GRIDS Nickel 100Vial 624.9590071 graticulesoptics
07DSPEC 2mm x 0.980mm HOLE GRIDS Copper with Rhodium 100Vial 693.94 graticulesoptics
07D02924 2mm x 0.980mm HOLE GRIDS Gold 25Vial 0 graticulesoptics
08L07924 2mm x 0.980mm HOLE GRIDS Titanium 25Vial 3068 graticulesoptics
08L05924 2mm x 0.980mm HOLE GRIDS Molybdenum 25Vial 1312.942613 graticulesoptics
08L04924 2mm x 0.980mm HOLE GRIDS Aluminium 25Vial 1341.427647 graticulesoptics
08L06924 2mm x 0.980mm HOLE GRIDS Stainless Steel 25Vial 1110.587984 graticulesoptics
07D00925 0.42mm SLOT GRIDS Copper 100Vial 532.5483532 graticulesoptics
07D01925 0.42mm SLOT GRIDS Nickel 100Vial 624.9590071 graticulesoptics
07D02925 0.42mm SLOT GRIDS Gold 25Vial 0 graticulesoptics
08L07925 0.42mm SLOT GRIDS Titanium 25Vial 3068 graticulesoptics
08L05925 0.42mm SLOT GRIDS Molybdenum 25Vial 1312.942613 graticulesoptics
08L04925 0.42mm SLOT GRIDS Aluminium 25Vial 1341.427647 graticulesoptics
08L06925 0.42mm SLOT GRIDS Stainless Steel 25Vial 1110.587984 graticulesoptics
07D00926 7 HEXAGON GRIDS Copper 100Vial 532.5483532 graticulesoptics
07D01926 7 HEXAGON GRIDS Nickel 100Vial 624.9590071 graticulesoptics
07D02926 7 HEXAGON GRIDS Gold 25Vial 878.7646285 graticulesoptics
03D00816 2mm x 980µm SLOTS Copper 100Vial 414.058056 graticulesoptics
03D00824 2mm x 980µm SLOTS Copper with Rhodium 100Vial 482.82 graticulesoptics
03D00817 2mm x 980µm SLOTS Nickel 100Vial 454.1376905 graticulesoptics
03D00818 0.2 x 1.5mm SLOTS Copper 100Vial 414.3086346 graticulesoptics
03D00819 0.2 x 1.5mm SLOTS Nickel 100Vial 454.1376905 graticulesoptics
03D00820 0.2 x 0.5mm SLOTS Copper 100Vial 414.3086346 graticulesoptics
03D00821 0.2 x 0.5mm SLOTS Nickel 100Vial 454.1376905 graticulesoptics
07D30918 0.4mm HOLE GRIDS Copper 100Vial 555.1300433 graticulesoptics
07D31918 0.4mm HOLE GRIDS Nickel 100Vial 694.1584771 graticulesoptics
07D30919 0.6mm HOLE GRIDS Copper 100Vial 555.1300433 graticulesoptics
07D31919 0.6mm HOLE GRIDS Nickel 100Vial 694.1584771 graticulesoptics
07D30920 0.8mm HOLE GRIDS Copper 100Vial 555.1300433 graticulesoptics
07D31920 0.8mm HOLE GRIDS Nickel 100Vial 694.1584771 graticulesoptics
07D30921 1.0mm HOLE GRIDS Copper 100Vial 555.1300433 graticulesoptics
07D31921 1.0mm HOLE GRIDS Nickel 100Vial 694.1584771 graticulesoptics
07D32921 1.0mm HOLE GRIDS Gold 25Vial 978.7732896 graticulesoptics
07D30924 2mm x  980µm SLOT GRIDS Copper 100Vial 555.1300433 graticulesoptics
07D31924 2mm x  980µm SLOT GRIDS Nickel 100Vial 694.1584771 graticulesoptics
07D32924 2mm x  980µm SLOT GRIDS Gold 25Vial 978.7732896 graticulesoptics
07D30925 0.42mm SLOT GRIDS Copper 100Vial 555.1300433 graticulesoptics
07D31925 0.42mm SLOT GRIDS Nickel 100Vial 694.1584771 graticulesoptics
07D30926 7 HEXAGON GRIDS Copper 100Vial 555.1300433 graticulesoptics
07D31926 7 HEXAGON GRIDS Nickel 100Vial 694.1584771 graticulesoptics
07D00932 100 MESH FINDERS Copper 100Vial 1193.556784 graticulesoptics
07D01932 100 MESH FINDERS Nickel 100Vial 1361.163959 graticulesoptics
07D02932 100 MESH FINDERS Gold 25Vial 1860.816891 graticulesoptics
07D00933 200 MESH FINDERS Copper 100Vial 1193.556784 graticulesoptics
07D01933 200 MESH FINDERS Nickel 100Vial 1361.163959 graticulesoptics
07D02933 200 MESH FINDERS Gold 25Vial 1860.816891 graticulesoptics
07D00965 200 MESH AHERA TYPE INDEX NEW Copper 100Vial 432.64 graticulesoptics
07D00966 200 MESH AHERA TYPE INDEX NEW Copper 100Vial 568.36 graticulesoptics
07D00934 300 MESH FINDERS Copper 100Vial 1315.299465 graticulesoptics
07D01934 300 MESH FINDERS Nickel 100Vial 1361.163959 graticulesoptics
07D02934 300 MESH FINDERS Gold 25Vial 1999.845324 graticulesoptics
07D00935 400 MESH FINDERS Copper 100Vial 1315.299465 graticulesoptics
07D01935 400 MESH FINDERS Nickel 100Vial 1508.695674 graticulesoptics
07D02935 400 MESH FINDERS Gold 25Vial 1999.845324 graticulesoptics
07D00940 200 MESH HIGH OPEN AREA Copper 100Vial 362.4346886 graticulesoptics
07D00941 400 MESH HIGH OPEN AREA Copper 100Vial 362.4346886 graticulesoptics
07D00942 75 MESH FREEZE FRACTURE Copper 100Vial 370.4774944 graticulesoptics
07D01942 75 MESH FREEZE FRACTURE Nickel 100Vial 416.4889252 graticulesoptics
07D02942 75 MESH FREEZE FRACTURE Gold 25Vial 611.5283702 graticulesoptics
07D00936 CHIEN GRID Copper 100Vial 568.8233959 graticulesoptics
07D00944 150 MESH TISSUE PROCESSING SCREEN DIAMETER 3.8mm Copper 100Vial 568.8233959 graticulesoptics
07D01944 150 MESH TISSUE PROCESSING SCREEN DIAMETER 3.8mm Nickel 100Vial 740.0157501 graticulesoptics
07D00945 200 MESH TISSUE PROCESSING SCREEN DIAMETER 3.8mm Copper 100Vial 740.0157501 graticulesoptics
07D01945 200 MESH TISSUE PROCESSING SCREEN DIAMETER 3.8mm Nickel 100Vial 740.0157501 graticulesoptics
07D00946 300 MESH TISSUE PROCESSING SCREEN DIAMETER 3.8mm Copper 100Vial 740.0157501 graticulesoptics
07D01946 300 MESH TISSUE PROCESSING SCREEN DIAMETER 3.8mm Nickel 100Vial 740.0157501 graticulesoptics
03D00600 H1 PATTERN, 200 MESH Copper 100Vial 401.0549327 graticulesoptics
03D00601 H1 PATTERN, 200 MESH Nickel 100Vial 440.1525323 graticulesoptics
03D00602 H1 PATTERN, 200 MESH Gold 100Vial 3186.859599 graticulesoptics
03D00620 H4 PATTERN, 400 MESH Copper 100Vial 401.0549327 graticulesoptics
03D00621 H4 PATTERN, 400 MESH Nickel 100Vial 440.5029588 graticulesoptics
03D00622 H4 PATTERN, 400 MESH Gold 100Vial 3188.886004 graticulesoptics
03D00606 H9 PATTERN, 50 MESH Copper 100Vial 389.2140351 graticulesoptics
03D00607 H9 PATTERN, 50 MESH Nickel 100Vial 420.3642738 graticulesoptics
03D00608 H9 PATTERN, 50 MESH Gold 100Vial 3069.328109 graticulesoptics
03D00822 HF52 PATTERN, 100 MESH Copper 100Vial 414.3086346 graticulesoptics
03D00823 HF52 PATTERN, 100 MESH Nickel 100Vial 454.1376905 graticulesoptics
03D00753 H2 REFERENCE PATTERN, 200 MESH LONDON FINDER Copper 100Vial 725.40537 graticulesoptics
03D00754 H2 REFERENCE PATTERN, 200 MESH LONDON FINDER Nickel 100Vial 832.5311163 graticulesoptics
03D00755 H2 REFERENCE PATTERN, 200 MESH LONDON FINDER Gold 100Vial 4879.438969 graticulesoptics
03D00759 H7 REFERENCE PATTERN, 400 MESH Copper 100Vial 725.40537 graticulesoptics
03D00760 H7 REFERENCE PATTERN, 400 MESH Nickel 100Vial 832.5311163 graticulesoptics
03D00761 H7 REFERENCE PATTERN, 400 MESH Gold 100Vial 5025.822138 graticulesoptics
03D00756 H6 REFERENCE PATTERN Copper 100Vial 725.40537 graticulesoptics
03D00757 H6 REFERENCE PATTERN Nickel 100Vial 832.5311163 graticulesoptics
03D00758 H6 REFERENCE PATTERN Gold 100Vial 4879.438969 graticulesoptics
03D00609 H12 FOLDED GRID/REFERENCE PATTERN Copper 25Vial 694.1584771 graticulesoptics
03D00632 H12 FOLDED GRID/REFERENCE PATTERN Nickel 25Vial 823.7471425 graticulesoptics
03D00631 H12 FOLDED GRID/REFERENCE PATTERN Copper with Rhodium 25Vial 862.42 graticulesoptics
03D00610 HF14 200 MESH FREEZE FRACTURE Copper with Rhodium 100Vial 408.72 graticulesoptics
03D00611 HF14 200 MESH FREEZE FRACTURE Nickel 100Vial 420.3642738 graticulesoptics
03D00612 HF14 200 MESH FREEZE FRACTURE Gold 100Vial 3069.328109 graticulesoptics
03D00750 HF15 REFERENCE PATTERN Copper with Rhodium 100Vial 725.40537 graticulesoptics
03D00751 HF15 REFERENCE PATTERN Nickel 100Vial 832.5311163 graticulesoptics
03D00752 HF15 REFERENCE PATTERN Gold 100Vial 5025.822138 graticulesoptics
07D30932 100 MESH FINDERS Copper 100Vial 1193.556784 graticulesoptics
07D31932 100 MESH FINDERS Nickel 100Vial 1361.163959 graticulesoptics
07D30933 200 MESH FINDERS Copper 100Vial 1193.556784 graticulesoptics
07D31933 200 MESH FINDERS Nickel 100Vial 1361.163959 graticulesoptics
07D32933 200 MESH FINDERS Gold 25Vial 1863.440069 graticulesoptics
07D30934 300 MESH FINDERS Copper 100Vial 1193.556784 graticulesoptics
07D31934 300 MESH FINDERS Nickel 100Vial 1361.163959 graticulesoptics
07D32934 300 MESH FINDERS Gold 25Vial 1863.440069 graticulesoptics
07D30935 400 MESH FINDERS Copper 100Vial 1193.556784 graticulesoptics
07D31935 400 MESH FINDERS Nickel 100Vial 1508.695674 graticulesoptics
07D32935 400 MESH FINDERS Gold 25Vial 2035.914022 graticulesoptics
07D00937 100 MESH COORDINATE Copper 100Vial 1193.556784 graticulesoptics
07D01937 100 MESH COORDINATE Nickel 100Vial 1378.807934 graticulesoptics
07D00938 200 MESH COORDINATE Copper 100Vial 1193.556784 graticulesoptics
07D01938 200 MESH COORDINATE Nickel 100Vial 1378.807934 graticulesoptics
07D00939 300 MESH COORDINATE Copper 100Vial 1193.556784 graticulesoptics
07D01939 300 MESH COORDINATE Nickel 100Vial 1378.807934 graticulesoptics
07D30942 75 MESH FREEZE FRACTURE Copper 100Vial 499.3875109 graticulesoptics
07D31942 75 MESH FREEZE FRACTURE Nickel 100Vial 555.1300433 graticulesoptics

 

 

EM Industries Inc 代理


EM Industries Inc 代理

简要描述:EM Industries Inc代理,EM Industries Inc上海代理,EM Industries Inc北京代理, EM Industries Inc代理, EM Industries Inc一级代理, EM Industries Inc 代理
上海金畔生物科技有限公司 EM Industries Inc专业代理,具体产品信息欢迎电询:021-50837765

详细介绍

产品咨询

世界*实验材料供应商 EM Industries Inc 上海金畔生物为其中国代理, EM Industries Inc 在一直是行业的*,一直为广大科研客户提供zui为优质的产品和服务,上海金畔生物一直秉承为中国科研客户带来的产品,的服务, EM Industries Inc 就是为了给广大科研客户带来更加完善的产品和服务,您的满意将是我们zui大的收获

 EM Industries Inc 中国代理, EM Industries Inc 上海代理, EM Industries Inc 北京代理,EM Industries Inc 广东代理, EM Industries Inc 江苏代理EM Industries Inc 湖北代理,EM Industries Inc 天津,EM Industries Inc 黑龙江代理,EM Industries Inc 内蒙古代理,EM Industries Inc 吉林代理,EM Industries Inc 福建代理, EM Industries Inc 江苏代理, EM Industries Inc 浙江代理, EM Industries Inc 四川代理,

 

EM Industries, is the US affiliate of Merck KGaA, Darmstadt, Germany. With sales in excess of $5 billion, Merck KGaA is a global supplier to the food, beverage and supplement industries. We are a leading producer of Hi-DHA Omega 3-Fatty Acids, Vitamins, Minerals and specialized forms of Sorbitol and Mannitol. 

 

上海金畔生物科技有限公司

5-diagnostics EM-101210说明书

5-diagnostics EM-101210说明书


5-diagnostics中国代理

 

Ref. No. EM-101210

Emo-test HIT Confirm®

Emo-test HIT Confirm® 通过流式细胞仪测量血小板活化,可以快速可靠地确认肝素诱导的血小板减少症 (HIT)。

HIT CONFIRM – 一种处理 HIT 测试的新方法

供体血小板的选择——对所有测试的挑战,对 HIT 的挑战更少
确认 HITT 在几分钟内确认,而不是几天
易用性 – 按需提供,24/7
使用微升,而不是毫升
质量和更好的标准化

尺寸 

20 次测试

SKU:EM-101210

 

HIT CONFIRM – 利用流式细胞术的力量 – 确认肝素诱导的血小板减少症 (HIT)

Emo-test HIT Confirm® 由 Emosis 开发,旨在满足临床医生和实验室获得 HIT 活动测试的需求,该测试将随时可用、按需、24/7、负担得起且易于使用专家。

HIT Confirm 测试可用于医院越来越多的流式细胞仪。新一代更小、更便宜的台式流式细胞仪,更易于非专业细胞仪使用,支持更多医院获得如此强大的技术。

Emo-test HIT Confirm® 是一种基于流式细胞术的 IVD 诊断试剂盒,旨在确认患者的 HIT 抗体是否存在并导致肝素-抗体-PF4 复合物的形成,该复合物将与供体血浆的血小板(富血小板血浆 – PRP),激活它们并对患者产生潜在的破坏性后果。

在血小板活化过程中,血小板膜会引起表面标志物表达的变化。这些血小板活化标志物可以在流式细胞术中使用荧光标记的配体进行研究。
HIT Confirm 测试仅使用一个步骤(一次孵育),两个荧光团针对血小板标记物 (CD 41) 和活化血小板标记物 (CD 62)。与 2 种水平的肝素(0.3 U/mL 和 100 U/mL)孵育 30 分钟后,通过流式细胞术(1 种激光,2 种颜色)计数 10,000 个事件获得相关活化血小板的比例。

基于从供体(用于 PRP)获得的血小板,使用称为 HEPLA(Emosis 开发)的血小板活化指数来解释结果,以获得更好的标准化结果

 

swissci EM-GRID-BOX-10说明书

swissci EM-GRID-BOX-10说明书

 

Cryo-EM

 

低温电磁

事实

  • 防静电聚合物
  • 提供的标签
  • 4个职位

主要特征

  • 新颖机制
  • 高性价比
  • 编号位置

 

 

关于

格形盒构成了Cryo-EM工作流程的重要组成部分,因为它们可以在液氮存储杜瓦瓶中将玻璃化的样品转移和存储在标准EM格上。SWISSCI Cryo EM Grid Box是专门为改善当前解决方案的许多功能而开发的。SWISSCI Cryo EM Grid Box是与剑桥MRC分子生物学实验室紧密合作开发的。SWISSCI Cryo EM网格盒由特殊的聚合物配方制成,可在低温下实现零静电。该盒子有四个菱形的插槽,带有识别号。由于盖子是用标准实验室镊子旋转的,因此不需要特殊工具。包装盒随附编号的压敏粘合低温标签。

产品规格

•防静电聚合物
•四个编号的菱形位置
•可选使用编号的压力粘合低温标签
•标签的位置允许在存储和使用过程中轻松查看标识号
•是液氮存储系统的理想选择
•标准圆形尺寸
•易于操作用镊子转动盖子
•易于使用且经济

订单信息

项目: Cryo EM Grid Box
代码: EM-GRID-BOX-10 EM-GRID-BOX-20 EM-GRID-BOX-50 
详细信息:每包10、20和50包  

 

 

mitegen Cryo-EM Pucks产品

mitegen Cryo-EM Pucks产品

品牌:mitegen

 

Cryo-EM Pucks

使用我们用于存储和跟踪低温EM网格盒的先进系统,在存储和运输过程中保持冷冻EM样品的安全和有序。

我们的Cryo-EM Pucks具有12个编号井,一个透明盖子和易于取用的镊子槽口,便于检索网格盒。每个冰球都可以使用仅有的序列号进行定制,以便您了解每个样品的位置,并为下一次显微镜访问时间做好准备。

MiTeGen Cryo-EM Pucks现在具有Lid Lock机制。这使得圆盘可以容易地装入存储和运输手杖中。请参阅下面的完整说明以了解更多信息。

在储存和运输过程中保持样品安全有序,使您的实验室能够专注于研究。我们设计了先进的系统,可以在低温条件下轻松跟踪Cryo-EM标本网格。

  • 每个圆盘有十二(12)个分度井
  • 井的顶部和底部的井数,便于查看
  • *的盖子锁定机制可以固定盖子,方便手杖装载

  • 持有圆形冷冻网格盒
  • 冰球深度适用于针式盖子式Cryo Grid Boxes
  • 当使用带有平面式盖子的Cryo Grid Box时,每个冰球多可存储24个Cryo Grid Box
  • 特殊的镊子插槽可以轻松安全地移除Cryo Grid Box
  • 搁架运输手杖多可容纳七个冰球(兼容CX100 Dryshipper)
  • Shelved Storage Cane可存放10个冰球(与HC34,HC35和VHC35存储杜瓦瓶兼容)

一套完整的Cryo-EM冰球和工具:

  • 7 – Cryo-EM Pucks
  • 1 – Shelled Puck运输手杖(兼容CX100)
  • 1 – 搁置式存储手杖(兼容HC34)
  • 1 – 斜角冷冻钳
  • 1 – 保护性存储盒
  • 每个冰球的定制雕刻

 

MiTeGen Cryo-EM Puck Starter Kit

SKU: M-CEM12-STK1

$2,397.00

MiTeGen Cryo-EM Pucks (standard, 12 hole) (free custom engraving available)SKU: M-CEM12-1

$105.00

Set of Seven (7) MiTeGen Cryo-EM Pucks (standard, 12 hole) (free custom engraving available)SKU: M-CEM12-7

$700.00

Shelved Puck Shipping CaneSKU: M-CP-111-065

$892.00

Bent Cryo-Tongs (for puck manipulation)SKU: M-CP-111-030

$115.00

Shelved Storage Cane (HC34 Compatible) holds 10 CryoEM pucksSKU: M-CEMSTC-1

$400.00

Protective Storage CaseSKU: M-CEMPSC-1

$290.00

Free Custom Engraving of Cryo-EM PucksSKU: M-CEM12-ENGRAVE

$0.00

Multi-Color Puck OptionSKU: M-CP-CustomColors

$0.00

HC34 High-Capacity Liquid Nitrogen Refrigerator (HC34)SKU: TW-HC34

$1,260.00

Cryo-EM Puck Dewar and InsertSKU: M-CEM-PD-IS1

$525.00

Categories:  Cryo-EMPucks

 

 

mitegen中国代理, mitegen上海代理, mitegen北京代理,mitegen广东代理, mitegen江苏代理mitegen湖北代理,mitegen天津代理, mitegen黑龙江代理, mitegen内蒙古代理, mitegen吉林代理, mitegen福建代理,  mitegen江苏代理, mitegen浙江代理,  mitegen四川代理,mitegen湖南代理,

 

上海金畔生物科技有限公司是实验试剂一站式采购服务商

1:强大的进口辐射能力,血清、抗体、耗材、大部分限制进口品等。

2:产品种类齐全,经营超过700多品牌,基本涵盖所有生物实验试剂耗材。

3:提供加急服务,货品一般1-2周到货。

4:富有竞争力的价格优势,绝大部分价格有优势。

5:多年积累良好的信誉,大部分客户提供货到付款服务。客户包括清华、北大、交大、复旦、中山等100多所高校,ROCHE,阿斯利康、国药、fisher等知药企。

6:我们还是Santa,Advanced Biotechnologies Inc,Athens Research & Technology,bangs,BBInternational,crystalchem,dianova,FD Neurotechnologies,Inc. FormuMax Scientific,Inc, Genebridege, Glycotope Biotechnology GmbH; iduron,Innovative Research of America, Ludger, neuroprobe,omicronbio, Polysciences,prospecbi, QA-BIO,quickzyme,RESEARCH DIETS,INC,sterlitech;sysy,TriLink BioTechnologies,Inc;worthington-biochem,zyagen等几十家国外公司代理。

7:我们还是invitrogen,qiagen,MiraiBioam,sigma;neb,roche,merck, rnd,BD, GE,pierce,BioLegend等知*批发,欢迎合作。

 

 

 

ESIBIO EM2202共培养分析方案

 

PROTOCOL

VascuNet™ Pericyte Co-Culture Assay

VascuNet™Pericyte共培养试验

 

INTRODUCTION 介绍

Vasculogenesis and angiogenesis are the mechanisms responsible for the development of a vascular network. Both endothelial cells and pericytes are key cell types in these processes. While endothelial cells form the lining of the vessels, pericytes are essential to the development of functional vasculature by stabilizing established vessel structures and facilitating local remodeling for network expansion. In addition to conveying structural support, pericytes are also integral in directing endothelial cells via cell-to-cell contact and paracrine signaling. Pericytes have been shown to co-localize with endothelial cells in both normal and abnormal vasculature, and have been implicated in playing a central role in numerous pathologies, including tumorigenesis, neurodegenerative disorders, and diabetic retinopathy.

血管生成和血管生成是血管网络发育的机制。内皮细胞和周细胞都是这些过程中的关键细胞类型。而内皮 细胞是血管的内层,周细胞通过稳定已建立的血管结构和促进网络e的局部重塑,对功能血管的发展至关重要。 xpansion。除了传递结构支持外,周细胞还通过细胞与细胞的接触和旁分泌信号来引导内皮细胞。周细胞已被证明为共室细胞。 在正常和异常血管系统中,内皮细胞与血管内皮细胞结合,并参与多种病理学中的中心作用,包括肿瘤发生、神经退行性疾病。 糖尿病视网膜病变。

 

The VascuNet Pericyte Co-Culture Assay combines human embryonic stem cell (ESI-017)-derived pericytes (PC-M cells) with primary human umbilical vein endothelial cells (HUVECs) in a co-culture system designed for a 96-well plate format. These unique PC-M cells display several key properties of pericytes, including expression of CD146, pro-angiogenic function, and effective stabilization of endothelial tube networks. The HUVECs and PC-M cell co- culture system supports vasculogenic tube assembly, resulting in the generation of extensive tube networks that persists at least 4 to 6 days in culture. Vasculogenic tube networks formed with the VascuNet Pericyte Co-Culture Assay persist over 4 times longer in culture than those formed by other assay systems, allowing researchers to study the relevant timing of delivery and long-term efficacy of pro- and anti-angiogenic compounds.

 

人胚胎干细胞(esi-017)衍生周细胞(pc-M细胞)与原代人脐静脉内皮细胞(HUVECs)在共培养系统中的结合。 设计的96井板格式。这些*的pc-M细胞显示了周细胞的几个关键特性,包括CD 146的表达、促血管生成功能以及内皮细胞的有效稳定。 光管网络。HUVECs和pc-M细胞共培养系统支持血管生成管的组装,从而产生了在Cultur至少持续4至6天的广泛的管状网络。 e.用VascuNet Pericyte共培养法形成的致血管网络在培养过程中的持续时间是其他检测系统的4倍以上,这使得研究人员可以研究该方法。 促血管生成化合物和抗血管生成化合物的时间安排和长期疗效。

IMPORTANT TIPS  重要提示

  • All work with live cells should be completed in a sterile biological safety cabinet designated for tissue culture. 所有活细胞的工作应在组织培养的无菌生物安全柜内完成。
  • Do not allow either PC-M cells or HUVECs to proliferate to more than 90% confluency during expansion. At high confluency, dense cell clusters may form, which can decrease vasculogenic tube assembly in mono- and co-cultures. Be sure to resuspend cells to single-cell suspension before plating for the angiogenic assay.不允许PC-M细胞或HUVECs在扩张过程中增殖到90%以上。在高度汇合的情况下,可形成密集的细胞团簇,从而减少血管生成管的组装。 单一文化和共同文化。在进行血管生成试验之前,一定要将细胞重新悬浮到单细胞悬浮液中.
  • Solvents used to dissolve test reagents, such as DMSO. may have inherent pro- or anti-angiogenic properties For this reason, all test reagents should be resuspended in diluents with no greater than 0.1% (v/v) of the solvent.用于溶解试验试剂的溶剂,如dmso,可能具有固有的亲或抗血管生成特性,因此,所有试验试剂应重新悬浮在不超过0的稀释剂中。 .1%(v/v)的溶剂。
  • Optimal vascular tube growth and stability is achieved when the HUVECs and PC-M cells are plated at a total of 42,000 cells per well in a ratio of 20:1 for HUVECs:PC-M cells, respectively.当HUVECs与PC-M细胞以42,000细胞/孔的比例分别以20:1的比例镀膜时,可获得宜的血管生长和稳定性。

 

REQUIRED MATERIALS所需材料

The VascuNet Pericyte Co-Culture Assay kit contains PC-M cells, HUVECs, and all media components for cell expansion and vasculogenic assay. Each kit undergoes extensive quality control to ensure reproducible vasculogenic tube assembly.

Vascune-周细胞共培养检测试剂盒含有PC-M细胞、HUVECs和所有用于细胞扩增和血管生成测定的培养基成分。每个套件都进行了全面的质量控制,以确保RPR。 可排卵的血管生成管组装。

 

VascuNet™ Pericyte Co-Culture Assay Kit Contents and Storage Conditions

 

Kit Component

Quantity

Storage Condition

Cells:

PC-M cells (ESI-017-derived pericytes) p19

1 vial

≥ 5.0 × 105 cells/vial

Liquid Nitrogen

HUVECs (secondary donor pool) p5

1 vial

≥ 1.0 × 106 cells/vial

Liquid Nitrogen

VascuNet Growth Medium Components:

VascuNet Basal Medium

475 mL

2 to 8°C

Recombinant Human VEGF

0.5 mL

-20°C

Recombinant Human EGF

0.5 mL

-20°C

Recombinant Human IGF-1

0.5 mL

-20°C

Recombinant Human FGF basic

0.5 mL

-20°C

Ascorbic Acid

0.5 mL

-20°C

Heparin Sulfate

0.5 mL

-20°C

Hydrocotrisone Hemisuccinate

0.5 mL

-20°C

FBS

25 mL

-20°C

L-Glutamine

25 mL

-20°C

VascuNet Assay Medium Components:

VascuNet Basal Assay Medium

95 mL

2 to 8°C

L-Glutamine

5 mL

-20°C

Negative Control:

Suramin Hexasodium Salt

0.5 mL,

1 mM in H2O

-20°C

 

Ensure that kit components are stored at the indicated temperatures upon kit arrival. The VascuNet Pericyte Co- Culture Assay components are stable for a minimum of 3 months from date of receipt when stored as directed.

确保组件在组件到达时以的温度存储。VascuNet Pericyte共同培养测试组件从收到时起至少3个月内是稳定的。 按指示存储。

 

 

 

Additional Required Reagents and Materials

 

 

附加所需试剂和材料

  • Sample test compounds to assay
  • 样品检测化合物
  • Corning® Matrigel® Growth Factor Reduced (GFR) Basement Membrane Matrix, *LDEV- Free
  • Corning Matrigel生长因子减少(GFR)基底膜基质,*LDEV-无
  • Phosphate Buffered Saline (PBS)
  • 磷酸盐缓冲盐(PBS)
  • Accutase® Cell Detachment Solution
  • 酸性磷酸酶细胞脱离液
  • Trypan blue or alternative cell viability assay
  • 台盼蓝或替代细胞活力测定
  • 0.22 µm sterile filtration unit, 500 mL 
  • 0.22毫升无菌过滤装置,500 mL
  • 0.22 µm sterile filtration unit, 150 mL
  • 0.22毫升无菌过滤装置,150 mL
  • T150 tissue culture flasks
  • T 150组织培养瓶
  • T75 tissue culture flasks
  • T75组织培养瓶
  • 96-well tissue culture plate

96孔组织培养板

EXPERIMENTAL OVERVIEW

 

Experimental Timeline

Assay Set-up

 实验时间线

 

Row 1: Control Row

H: HUVEC Monoculture Reference (40,000 cells/well)

P: PC-M Monoculture Reference (2,000 cells/well)

C: Co-Cultures (20:1 ratio of HUVEC:PC-M; 40,000 HUVECs and 2,000 PC-M cells/well)

Blue: HUVEC & PC-M Cell Co-Culture (Positive Control)

Yellow: HUVEC & PC-M Cell Co-Culture with 50 µM Suramin Hexasodium Salt (Negative Control)

 

第1行:控制行

 

h:HUVEC单一培养参考资料(40,000个细胞/井)

 

P:PC-M单细胞培养参比(2,000个细胞/井)

 

C:共培养(20:1的HUVEC:PC-M;40,000 HUVECs和2,000个PC-M细胞/井)

 

蓝:HUVEC&PC-M细胞共培养(阳性对照)

 

黄:HUVEC&PC-M细胞与50 M苏拉明六钠盐共培养(阴性对照

 

Figure 1. Example assay set-up in a 96-well plate. The first row of the VascuNet Pericyte Co-Culture Assay is a control row, containing triplicate samples of monoculture wells and positive and negative control co-culture wells. The remaining 84 wells of the plate may be used for test compounds. Test compound assays may be performed following the same mono- and co-culture set-up as the control row, or simply as co-culture wells.

 

图1.在一个96井的平板上进行测试。VascunetPericyte共培养试验的行是对照行,包含三份单一培养井的样本以及阳性和阴性的样本。 控制共培养井。该板的其余84口井可用作试验化合物。测试复合测试可以按照与控制行相同的单文化和共培养设置执行。 或者简单地说是共文化水井。

 

 

EXPERIMENTAL PROTOCOL

 

 

 

EXPANSION OF HUVECs AND PC-M CELLS

HUVECs和PC-M细胞的扩增

The HUVECs and PC-M cells must be thawed and expanded individually for two days before they can be plated for the experimental set-up in a co-culture format.

Monitor cell proliferation, and do not allow either the HUVECs or PC-M cells to proliferate to more than 90% confluency during expansion. At high confluency dense cell clusters may form, which can decrease vasculogenic tube assembly in mono- and co-cultures.

HUVECs和PC-M细胞必须单独解冻和扩张两天,然后才能以共同培养的形式进行实验设置。

 

监测细胞增殖,不允许HUVECs或PC-M细胞在扩张过程中增殖至90%以上。在高汇合度时,可形成密集的细胞团簇。 单培养和共培养时血管生成管的组装。

 

DAY -4

 

Prepare VascuNet Growth Medium for HUVECs and PC-M Cell Expansion

 

  1. Remove the following VascuNet Growth Medium supplements from -20°C storage and allow the following reagents to thaw at 2 to 8°C:

rhVEGF, rhEGF, rhIGF-1, rhFGF basic, Ascorbic Acid, Heparin Sulfate, Hydrocortosone Hemisuccinate, FBS, L-Glutamine

HUVECs和PC-M细胞扩增用VascuNet生长培养基的制备

 

将以下VascuNet生长介质从-20℃储存中移除,并允许下列试剂在2至8℃解冻:

 

rhVEGF,rhEGF,rhIGF-1,rhFGF碱性,抗坏血酸,硫酸肝素,半琥珀酸氢皮质酮,FBS,L-谷氨酰胺

  1. Prepare VascuNet Growth Medium by combining all the reagents listed below.

VascuNet Basal Medium 475 mL

rhVEGF 0.5 mL

rhEGF 0.5 mL

rhIGF-1 0.5 mL

rhFGF basic 0.5 mL

Ascorbic Acid 0.5 mL

Heparin Sulfate 0.5 mL Hydrocotrisone Hemisuccinate 0.5 mL FBS 25 mL

  • Glutamine 25 mL

通过结合下面列出的所有试剂,准备VascuNet生长培养基。

 

VascuNet Basal培养基475 mL

 

rhVEGF 0.5 mL

 

rhEGF 0.5 mL

 

rhIGF-1 0.5 mL

 

rhFGF碱性0.5 mL

 

抗坏血酸0.5mL

 

硫酸肝素0.5 mL氢曲松半琥珀酸0.5 mL FBS 25 mL

 

L-谷氨酰胺25 mL

 

  1. Filter sterilize the VascuNet Growth Medium using a 0.22 µm pore size, low protein-binding filter unit into a sterile 500 mL bottle.滤器用0.22m孔径、低蛋白质结合的过滤装置消毒VascuNet培养基,制成无菌500 mL瓶。
  2. Transfer 75 mL of the VascuNet Growth Medium to a sterile 100 mL bottle and place in a 37°C water bath for 30 minutes to warm.将75 mL的VascuNet培养基转移到无菌的100 mL瓶中,置于37°C水浴中30分钟取暖。
  3. Store the remaining VascuNet Growth Medium at 2 to 8°C for up to 2 weeks.将剩余的VascuNet培养基保存在2至8°C处,多2周。

 

 

Thaw and Plate HUVECs for Expansion 用于膨胀的解冻板HUVECs

  1. Thaw the vial of HUVECs briefly in a 37°C water bath.将HUVECs瓶在37°C水浴中短暂解冻。
  2. Transfer the entire volume of the vial into 4 mL of pre-warmed VascuNet Growth Medium in a 15 mL conical tube.将整个体积的小瓶转移到4毫升的预热VascuNet生长培养基中,放入15 mL的锥形管中。

 

  1. Centrifuge the cell suspension for 5 minutes at 200 x g. 200×g离心细胞悬液5分钟。

 

  1. Aspirate the supernatant and gently resuspend the cell pellet in 10 mL of VascuNet Growth Medium.

 

吸出上清液,在10 mL的VascuNet培养基中轻轻悬浮细胞颗粒。

 

 

 

 

 

  1. Count the total number of viable cells using Trypan blue.使用台盼蓝计算活细胞总数。
  2. Add 10 mL of VascuNet Growth Medium to each of two T150 flasks.在两个T 150瓶中各加入10 mL VascuNet生长培养基。
  3. Divide the HUVEC suspension evenly between the two T150 flasks, such that each flask contains approximately 3.75 to 4.5 × 105 HUVECs at a density of 2.5 to 3.0 × 103 HUVECs/cm2.将HUVEC悬浮液均匀地分成两个T 150瓶,使每个瓶在2.5~3.0×103HUVECs/cm2的密度范围内含有约3.75~4.5×105个HUVECs。
  4. Add a sufficient volume of the VascuNet Growth Medium to each flask to bring the total volume to 30 mL per T150 flask.在每个瓶中加入足够量的VascuNet生长培养基,使总体积达到每T 150瓶30 mL。

 

  1. Incubate the cells overnight at 37°C with 5% CO2 humidified atmosphere.细胞在37°C和5%CO2加湿气氛中过夜

 

 

DAY -2

 

Thaw and Plate PC-M Cells for Expansion 用于膨胀的解冻板PC-M电池

  1. Transfer 45 mL of VascuNet Growth Medium to a sterile 50 mL conical tube and warm in a 37°C water bath for 30 minutes.将45 mL的VascuNet培养基转移到无菌的50 mL锥形管中,在37°C水浴中加热30 min。
  2. Thaw the vial of PC-M cells briefly in a 37°C water bath. Transfer the entire contents of the vial into 4 mL of pre-warmed VascuNet Growth Medium in a 15 mL conical tube.将PC-M细胞在37°C水浴中解冻。将小瓶的全部内容转移到4mL预加热的VascuNet生长培养基中,放入15 mL的锥形管中。
  3. Centrifuge the cell suspension for 5 minutes at 200 x g.200×g离心细胞悬液5分钟。
  4. Aspirate the supernatant and gently resuspend the cell pellet in 10 mL of VascuNet Growth Medium.吸出上清液,在10 mL的VascuNet培养基中轻轻悬浮细胞颗粒。

 

  1. Count the total number of viable cells using Trypan blue.使用台盼蓝计算活细胞总数。
  2. Add 10 mL of VascuNet Growth Medium to each of two T75 flasks.将10毫升VascuneT培养基加入到两个T75烧瓶中。
  3. Divide the PC-M cell suspension evenly between the two T75 flasks, such that each flask contains approximately 2.5 × 105 PC-M cells at a density of 1.0 × 104 PC-M cells/cm2.将PC-M细胞悬浮液均匀地划分到两个T75瓶之间,使每瓶容量约为2.5×105个PC-M细胞,密度为1.0×104pC-M细胞/cm2。

Note: The PC-M cells are plated at a higher density than the HUVECs.注:PC-M细胞的密度高于HUVECs.

  1. Add a sufficient volume of the VascuNet Growth Medium to each flask to bring the total volume to 15 mL per T75 flask.在每个瓶中加入足够量的VascuNet生长培养基,使总体积达到每T75瓶15 mL。
  2. Incubate the cells overnight at 37°C with 5% CO2 humidified atmosphere.细胞在37°C和5%CO2加湿气氛中过夜。

 

 

Exchange Medium in HUVEC Culture HUVEC培养中的交换培养基

  1. Warm 60 mL of VascuNet Growth Medium at 37°C.37°C温60 mL VascuNet培养基。
  2. Observe HUVEC cultures under microscope to assess cell proliferation, confluency, and overall health.显微镜下观察HUVEC培养,观察细胞增殖、融合及整体健康状况。
  3. Aspirate the medium in both of the T150 culture flasks, and replace with 30 mL of fresh VascuNet Growth Medium per flask.在T150培养瓶中抽吸培养基,每瓶更换30毫升新鲜VascuneT培养基。
  4. Incubate cells at 37°C with 5% CO2 humidified atmosphere.

37°C条件下,5%CO2加湿培养细胞

 

ANGIOGENESIS ASSAY 血管生成试验

Once the HUVECs and PC-M cells have been expanded, the HUVEC and PC-M cells are plated in co-cultures and monoculture wells of a 96-well plate for the angiogenesis assay. Experimental and control assay wells are established to determine the effects of test compounds on angiogenesis.一旦HUVECs和PC-M细胞被扩增,HUVEC和PC-M细胞被镀在96孔板的共培养和单培养井中进行血管生成实验。试验与控制驴 我们建立了测试井,以确定试验化合物对血管生成的影响。

 

 

DAY 0

 

Prepare 96-well Assay Plate制备96井试井板

 

  1. Thaw sufficient amounts of Growth Factor Reduced (GFR) Matrigel aliquot(s) on ice at 2 to 8°C.在冰上解冻足够数量的生长因子减少(GFR)Matrigel ali“(S)在2到8°C。

Note: Thaw a sufficient amount of GFR Matrigel to coat the total amount of wells used for the experiment, including the control row (see Fig. 1). Each well used in the assay will require 50 µL of Matrigel solution.注:融化足够数量的GFR Matrigel,以覆盖总数量的井用于实验,包括控制排(见图1)。在分析中使用的每一口井都需要50升的MAT。 Rigel溶液

  1. Place a 96-well tissue culture plate and P100 pipet tips at -20°C for 1 hour to chill.放置96孔组织培养板和P 100管尖,温度-20℃,冷藏1小时。
  2. Transfer the chilled 96-well plate, chilled pipet tips, and thawed GFR Matrigel bottle to an ice bucket in the tissue culture hood.将冰镇的96井板、冰镇管尖和融化的GFR Matrigel瓶转移到组织培养罩中的冰桶中。

Note: The GFR Matrigel will solidify rapidly when warmed above 2 to 8°C. To ensure even coating and distribution of the matrix, keep all of the reagents, tips, and plate on ice.注:GFR Matrigel在温度高于2°~8°C时会迅速凝固,以保证基体的均匀涂层和分布,使所有试剂、针尖和平板保持在冰上。

  1. Add 50 µL of GFR Matrigel to each well of the 96-well plate, changing tips often for even plating.在96井板的每口井中加入50升GFR Matrigel,经常改变镀液的。
  2. Incubate the coated 96-well plate at room temperature for 1 hour, then transfer to a 37°C incubator with 5% CO2 humidified atmosphere for 1 to 2 hours prior to plating cells.将包覆的96孔板在室温下孵育1h,再在37℃、5%CO2加湿气氛下孵育1~2小时。

 

 

Prepare VascuNet Assay Medium制备VascuNet检测介质

  1. Thaw the L-Glutamine (for assay medium) at 2 to 8°C. Allow the VascuNet Basal Assay Medium to warm to room temperature.将L-谷氨酰胺(用于检测介质)在2到8°C解冻。允许VascuNet Basal分析介质加热到室温。
  2. Prepare the VascuNet Assay Medium by combining the reagents below.通过组合下面的试剂,准备VascuNet测试介质。

VascuNet Basal Assay Medium 95 mL  VascuNet Basal试验培养基95 mL

L-Glutamine 5 mL   L-谷氨酰胺5mL

 

  1. Filter sterilize the VascuNet Assay Medium using a 0.22 µm pore size, low protein-binding filter and a sterile 100 mL bottle. 滤池用0.22m孔径、低蛋白质结合过滤器和无菌100 mL瓶对VascuNet分析培养基进行消毒。
  2. Transfer 50 mL of VascuNet Assay Medium to a 50 mL conical tube. Warm this aliquot in a 37°C water bath for 30 minutes. The required amount of medium may vary depending on the number of test component wells.将50 mL VascuNet试剂盒转入50 mL锥形管。在37℃的水浴中加热30分钟。所需介质的数量可能会根据测试组件的数量而有所不同。
  3. Store the remaining VascuNet Assay Medium at 2 to 8°C for up to 2 weeks.

将剩余的VascuNet检测介质保存在2至8°C处,多2周。

 

 

 

 

 

 

Harvest HUVECs  收获HUVECs

  1. Aspirate the culture medium from each of the T150 flasks, and add 10 mL of PBS per flask to wash. Aspirate the PBS wash.分别从T 150瓶中抽吸培养基,每瓶加入10 mL PBS进行洗涤。吸入PBS洗涤液。
  2. Add 5 mL of Accutase Cell Detachment Solution to each flask and incubate at room temperature for 5 minutes, or until cells appear rounded.在每瓶中加入5毫升酸性磷酸酶细胞脱落液,在室温下孵育5分钟,或直至细胞呈圆形。
  3. Add 5 mL of VascuNet Assay Medium to each flask. Firmly tap the side of the flask to release the cells from the culture surface.在每个瓶中加入5毫升VascuNet分析培养基。牢固地敲击瓶的侧面,将细胞从培养表面释放出来。
  4. Collect the HUVECs from each flask and transfer the cells within each flask to a 15 mL conical tube. Pipet gently to dissociate any remaining cell clumps.从每个烧瓶收集HUVECs,并将每个烧瓶内的细胞转移到15毫升锥形管中。轻轻吸管以分离的任何剩余的细胞团。
  5. Centrifuge the cells for 5 minutes at 250 x g.250×g离心5分钟。
  6. Aspirate the supernatant from each tube and resuspend the cell pellets to a single cell suspension in 5 mL of VascuNet Assay Medium per tube.每管抽取上清液,再将细胞颗粒悬浮到单个细胞悬液中,每管5 mL VascuNet检测培养基。

Note: It is essential to thoroughly resuspend the cells to single-cell suspension before plating for the angiogenic assay.注:在进行血管生成试验前,必须将细胞*重新悬浮到单细胞悬浮液中。

  1. Combine single cell suspensions (10 mL total volume) and pipet to mix.将单细胞悬液(总体积10 mL)与吸管混合。
  2. Count the total number of viable cells in the combined cell suspension using Trypan blue.用台盼蓝计数组合细胞悬液中的活细胞总数。
  3. Dilute the HUVECs in VascuNet Assay Medium to a concentration of 5 x 105 viable cells/mL.在VascuNet检测培养基中稀释HUVECs至5×105个活细胞/mL。

 

 

 

Harvest PC-M Cells 收获PC-M细胞

  1. Aspirate the culture medium from each of the T75 flasks, and add 5 mL of PBS per flask to wash. Aspirate the PBS wash.分别从T75瓶中抽吸培养基,每瓶加入5 mL PBS进行洗涤。吸入PBS洗涤液。
  2. Add 2.5 mL of Accutase Cell Detachment Solution to each flask and incubate at room temperature for 5 minutes, or until cells appear rounded.每瓶加入2.5mL的酸性磷酸酶细胞脱落液,在室温下孵育5分钟,直至细胞呈圆形。
  3. Add 2.5 mL of VascuNet Assay Medium to each flask. Firmly tap the side of the flask to release the cells from the culture surface.在每个瓶中加入2.5mL的VascuNet分析培养基。牢固地敲击瓶的侧面,将细胞从培养表面释放出来。
  4. Collect the PC-M cells from each flask and transfer to a single 15 mL conical tube. Pipet gently to dissociate any remaining cell clumps.从每个烧瓶中收集PC-M细胞,转移到一个15 mL的锥形管中。轻轻地将任何剩余的细胞团分离。
  5. Centrifuge cells for 5 minutes at 250 x g.250×g离心细胞5 min。
  6. Aspirate the supernatant and resuspend the cell pellet to a single cell suspension in 10 mL of VascuNet Assay Medium.取上清液,在10 mL VascuNet检测培养基中,将细胞颗粒再悬浮于单个细胞悬液中。

Note: It is essential to thoroughly resuspend the cells to single-cell suspension before plating for the angiogenic assay.注:在进行血管生成试验前,必须将细胞*重新悬浮到单细胞悬浮液中。

  1. Measure the total number of viable cells in the sample by performing a cell count using Trypan blue.通过使用台盼蓝执行细胞计数来测量样本中可行的细胞总数。
  2. Dilute the PC-M cells in VascuNet Assay Medium to a concentration of 1 x 105 viable cells/mL.

在VascuNet检测培养基中稀释PC-M细胞,浓度为1×105个活细胞/mL。

 

 

 

 

 

 

Plate HUVECs and PC-M Cells平板HUVECs和PC-M细胞

 

Refer to sample plate set-up diagram (Fig. 1). In addition to the experimental wells, one row of the 96-well plate will be utilized for control wells. Determine the total number of cells needed, in addition to the control row, based on the number of test samples.参考样板设置图(图1).除试验井外,96口井板中的一行井将用于控制井。确定所需单元格的总数,在 到控制行,根据测试样本的数量。

 

  1. For each set of triplicate HUVEC monoculture samples, combine 1.32 x 105 HUVECs with a sufficient volume of VascuNet Assay Medium to bring the total volume to 495 µL per triplicate set.对于每一组三重的HUVEC单培养样品,将1.32×105 HUVEC与足够体积的Vascuneta试验培养基相结合,使总体积为每三套495 L。
  2. Dispense 150 µL of the HUVEC cell suspension into each well of the assay, for a total of 4.0 x 104 HUVECs per monoculture well.将150 L的HUVEC细胞悬液分装于每口培养井中,平均每孔培养4.0×104个HUVECs。
  3. For each set of triplicate PC-M monoculture samples, combine 6.6 x 103 PC-M cells with a sufficient volume of VascuNet Assay Medium to bring the total volume to 495 µL per triplicate set.
  4. Dispense 150 µL of the PC-M cell suspension into each well of the assay, for a total of 0.2 x 104 PC-M cells per monoculture well.
  5. For each set of triplicate co-culture samples, combine 1.32 x 105 HUVECs with 6.6 x 103 PC-M cells. Add a sufficient volume of VascuNet Assay Medium to bring the total volume to 495 µL per triplicate set.对于每组三份PC-M单细胞培养样品,将6.6×103个PC-M细胞与足够体积的VascuNet检测培养基结合,使总体积达到每三份样品495 L。

Note: This will make a 20:1 ratio of HUVEC:PC-M cells.注:这将使HUVEC:PC-M细胞的比例达到20:1.

  1. Dispense 150 µL of the combined cell suspension into each well of the assay, for a total of 4.2 x 104 cells per co-culture well.将150 L的PC-M细胞悬液注入每孔,每孔培养一次,共培养0.2×104个PC-M细胞。
  2. Incubate the plate at 37°C with 5% CO2 humidified atmosphere undisturbed for 4 to 6 hours to allow cells to adhere and initial tube networks to form.每组三份共培养样品,将1.32×105个HUVECs与6.6×103个PC-M细胞结合.加入足够量的VascuNet检测介质,使总体积达到每三份495升 部分,段

 

 

Addition of the Test Compounds添加试验化合物

Following the 4 to 6 hour incubation period, visualy inspect the wells to confirm attachment and tube formation in both HUVEC monocultures and co-culture conditions (see Fig. 2). After confirmation of initial tube formation,

the cells are ready to be treated with the test reagents. For best results, this is done 4 to 6 hours after plating, but must be completed within 24 hours after cells are plated.在4到6小时的潜伏期后,目视检查水井,以确定在HUVEC单细胞培养和共培养条件下的附着和管状形成(见图2)。确认后 初的管状,这些细胞已准备好用试验试剂处理。为了取得良好的效果,这是在电镀后4至6小时,但必须在24小时内完成电池被镀。

 

Figure 2. VascuNet HUVEC and PC-M Cell Co-Culture at 4 hours. Co-Culture of HUVEC and PC-M cells seeded at a 20:1 ratio begin formation of tube networks as early as 4 hours.

图2.VascuNet HUVEC和PC-M细胞共培养4小时.人脐静脉内皮细胞(HUVEC)与PC-M细胞按20:1比例共培养,4小时开始形成管状网络。

 

 

 

 

 

 

Positive Control Wells 正控井

Positive control wells contain cells in co-culture at a 20:1 (HUVEC:PC-M cell) ratio in VascuNet Assay Medium. No additional compounds are added to these wells.阳性对照井在VascuNet检测培养基中含有20:1(HUVEC:PC-M细胞)比例的细胞。在这些井中不添加任何额外的化合物。

 

Negative Control Wells负压井

The negative control samples are run in triplicate co-culture conditions with 50 µM Suramin Hexasodium Salt.阴性对照样品一式三份,共培养条件为50 m苏拉明六钠盐。

 

  1. Thaw the vial of Suramin Hexasodium Salt for 1 hour at room temperature.在室温下解冻苏拉明六钠盐1小时。Add 25 µL of 1 mM Suramin Hexasodium Salt to 475 µL of VascuNet Assay Medium, for a final Suramin Hexasodium Salt concentration of 50 µM. Warm the 50 µM Suramin Hexasodium Salt solution to 37°C before use.在475升VascuNet分析介质中加入25升1毫米苏拉明六钠盐,使终的六钠浓度为50 M,然后将50 M苏拉明六钠盐溶液加热至37°C。 e使用。
  2. Aspirate the VascuNet Assay Medium from each well of cells and replace with 150 µL per well of the 50 µM Suramin Hexasodium Salt solution.从每口细胞中抽取VascuNet检测介质,用50 M苏拉明六钠盐溶液中的每口150毫升代替。

 

Experimental Wells 实验井

Experimental wells should be tested in co-culture wells in triplicates. Triplicate monoculture samples can also be run for comparison.实验威尔斯应在共培养威尔斯中试验三次。三份的单一栽培样品也可用于比较。

 

  1. Prepare additional pro- or anti-angiogenic test compounds by diluting with VascuNet Assay Medium to desired concentrations. Warm diluted test compound solutions to 37°C before use.用VascuNet分析培养基稀释至所需浓度,制备额外的亲或抗血管生成试验化合物。使用前将温热稀释的复合溶液稀释至37°C。
  2. Aspirate VascuNet Assay Medium from each well to be assayed and replace with 150 µL per well of VascuNet Assay Medium containing the appropriate test compound in solution.从每口井中抽吸VascuNet检测介质,用含适当试液的VascuNet分析介质每井150 L代替。

Day 1 to Day 4+ 第1天到第4天

 

  1. Observe and image the cells every 4 to 24 hours to monitor vasculogenic tube assembly and stability.

Note: Vascular tube formations in co-cultures containing both HUVECs and PC-M cells will be stable for at least 4 days in culture without the need to replace media or add exogenous factors.每4至24小时观察和成像细胞,以监测血管生成管的组装和稳定性。

 

注:在含有HUVECs和PC-M细胞的共培养中,血管管状细胞在培养中至少稳定4天,不需要更换培养基或添加外源因子。

 

EXPECTED RESULTS预期结果

The following data describes the expected results for co-culture and monoculture conditions at the recommended cell seeding density of 42,000 cells per well containing a 20:1 ratio of HUVECs to PC-M cells (40,000 HUVECs and 2,000 PC-M cells).以下数据描述了在建议的细胞播种密度为每井42,000个细胞时,共培养和单一培养条件下的预期结果,其中含有20:1的HUVECs与pc-m的比例。 细胞(40,000 HUVECs和2,000个PC-M细胞).

  • HUVEC monocultures will begin to form tube networks as early as 4 hours. A complete tube network can be observed at 24 hours. HUVEC tubes lacking PC-M cell support should destabilize by Day 2 of the angiogenesis assay and do not re-assemble.HUVEC单细胞早可在4小时内形成管状网络。24小时可观察到完整的管状网络。缺乏pc-M细胞支持的HUVEC管应在第2天发生不稳定。 血管生成实验和不重新组装。

 

  • PC-M cell monocultures do not form complete tube networks. Minimal branching may be observed.PC-M细胞不形成完整的管状网络.可以观察到小分支。

 

  • Co-cultures form tube networks within four days and are maintained for up to 6 days.共培养在4天内形成管状网络,并维持6天

 

  • The addition of 50 µM Suramin Hexasodium Salt (negative control) to both monocultures and co- cultures will reduce tube formation by > 90% within 4 to 24 hours after treatment. The presence of Suramin Hexasodium Salt will prevent tube re-assembly for at least 4 days.在单一培养和共培养中加入50μm苏拉明六钠盐(阴性对照),可在处理后4~24小时内使试管形成减少90%以上。苏拉米的存在 N六钠盐可以防止管重新组装至少4天。

 

Image Timeline

Figure 3. Stability of tube structures over time. HUVEC monocultures (rows 1 and 2) seeded at 120,000 cells/cm2 and stained with Vybrant DiO (green), show tube formation on Day 1 post seeding. By day 2, degredation of the vessels is already observed. In contrast, tube structures with multiple branching points are visible from Day 1 and are still stable at Day 6 in the HUVEC and PC-M cell co-cultured wells plated at a 20:1 ratio (rows 3 and 4). PC-M cells are stained red with Vybrant Dil.

Images were taken at 4X magnification.

图3.随着时间的推移,管状结构的稳定性。HUVEC单眼(第1行和第2行)在12万个细胞/cm2下播种,用VybrantDio(绿色)染色,在播种后第1天出现管状形成。白天 2、已观察到船舶的高度。相比之下,具有多个分支点的管状结构从第1天就可以看到,在HUVEC和pc-M细胞共培养中,在第6天仍保持稳定。 红色水井按20:1比例镀制(第3和第4行)。PC-M细胞用Vybrant Dil染红。

 

图像以4X放大倍数拍摄。

 

 

 

 

 

 

 

APPENDIX

 

COMMERCIAL SOURCES:商业来源:

 

Reagent

Source

Catalog Number

VascuNet Pericyte Co-Culture Assay

ESI BIO

EM-2202

Corning® Matrigel® Growth Factor Reduced (GFR) Basement

Membrane Matrix, *LDEV-Free

 

Corning

 

354230

Accutase® Cell Detachment Solution

Innovative Cell Technologies

AT104 – 100 mL

 

 

REFERENCES

 

Armulik, A., et al. (2005) Endothelial/pericyte interactions. Circ Res 97: 512–523.

Benjamin, L.E., et al. (1998) A plasticity window for blood vessel remodeling is defined by pericyte coverage of the preformed endothelial network and is regulated by PDGF-B and VEGF. Development 125: 1591–1598.

Bergers, G., et al. (2003) Benefits of targeting both pericytes and endothelial cells in the tumor vasculature with kinase inhibitors. J Clin Invest 111: 1287–1295.

Blocki, A., et al. (2013) Not all MSCs can act as pericytes: functional in vitro assays to distinguish pericytes from other mesenchymal stem cells in angiogenesis. Stem Cells Dev 22: 2347–2355.

Gerhardt, H., and Betsholtz, C. (2003) Endothelial-pericyte interactions in angiogenesis. Cell Tissue Res. 314: 15–23.

Hamilton, N.B., et al. (2010) Pericyte-mediated regulation of capillary diameter: a component of neurovascular coupling in health and disease. Front Neuroenergetics 2: 5.

Stratman, A.N., et al. (2009) Pericyte recruitment during vasculogenic tube assembly stimulates endothelial basement membrane matrix formation. Blood 114: 5091–5101.

von Tell, D., et al. (2006) Pericytes and vascular stability. Exp Cell Res 312: 623–629.

Zimmerlin, L., et al. (2010) Stromal vascular progenitors in adult human adipose tissue. Cytometry A 77A: 22–30.

参考( reference的名词复数 )

 

阿穆利克,A,等。(2005)内皮细胞/周细胞相互作用。CIRC RES 97:512—523。

 

本杰明,L.E.,等人。(1998)血管重塑的可塑性窗口是由预先形成的内皮网络的周细胞覆盖来定义的,由PDGF-B和VEGF调节。发展125 : 1591–1598.

 

Bergers,G.等人.(2003)用激酶抑制剂靶向肿瘤血管中的周细胞和内皮细胞的好处。j克莱因投资111:1287-1295。

 

Blocki,A.,等人。(2013)并非所有骨髓间充质干细胞都能作为周细胞:体外功能试验来区分周细胞和其他间充质干细胞在血管生成方面的作用。干细胞研发22:2347-2355。

 

Gerhardt,H.和Betsholtz,C.(2003)内皮-周细胞在血管生成中的相互作用。细胞组织研究314:15-23。

 

汉密尔顿,N.B.,等。(2010)围生体介导的毛细血管直径调节:健康和疾病中神经的血管耦合的一个组成部分。前神经能量学2:5。

 

施特拉曼,A.N.,等。(2009)血管生成管组装过程中围生细胞的招募刺激内皮基底膜基质的形成。血液114:5091-5101。

 

冯泰尔,D.(2006)周细胞和血管稳定性。实验细胞RES 312:623-629。

 

Zimulin,L,等。(2010)成人脂肪组织间质血管祖细胞。细胞计数A 77:22 – 30。

Matrigel is a registered trademark of Corning, Inc.

Accutase is a trademark of Innovative Cell Technologies, Inc.

© ESI BIO 2015. Unless otherwise noted, ESI BIO, ESI BIO logo and all other trademarks are the property of BioTime, Inc

 

 

ESI BIO产品目录:

 

货号

品名

规格

品牌

GS310

HyStem® Hydrogel Kit

2.5 mL

ESIBIO

GS311

HyStem® Hydrogel Kit

7.5 mL

ESIBIO

GS1004

HyStem® Hydrogel Kit

12.5 mL

ESIBIO

GS310P

HyStem® Hydrogel Kit w/ PEGSSDA

2.5 mL

ESIBIO

GS311P

HyStem® Hydrogel Kit w/ PEGSSDA

7.5 mL

ESIBIO

GS312

HyStem®-C Hydrogel Kit

2.5 mL

ESIBIO

GS313

HyStem®-C Hydrogel Kit

7.5 mL

ESIBIO

GS1005

HyStem®-C Hydrogel Kit

12.5 mL

ESIBIO

GS312P

HyStem®-C Hydrogel Kit w/ PEGSSDA

2.5 mL

ESIBIO

GS313P

HyStem®-C Hydrogel Kit w/ PEGSSDA

7.5 mL

ESIBIO

GS314

HyStem®-HP Hydrogel Kit

2.5 mL

ESIBIO

GS315

HyStem®-HP Hydrogel Kit

7.5 mL

ESIBIO

GS1006

HyStem®-HP Hydrogel Kit

12.5 mL

ESIBIO

GS314P

HyStem®-HP Hydrogel Kit w/ PEGSSDA

2.5 mL

ESIBIO

GS315P

HyStem®-HP Hydrogel Kit w/ PEGSSDA

7.5 mL

ESIBIO

GS1007

HyStem® Hydrogel UV QuickSet Kit

2.5 mL

ESIBIO

GS1008

HyStem® Hydrogel UV QuickSet Kit

7.5 mL

ESIBIO

GS450

PEGgel Kit

1 mL

ESIBIO

GS240

DG Water

10 mL

ESIBIO

GS241

DG Water

20 mL

ESIBIO

GS3007

Extralink® Vial

0.5 mL

ESIBIO

GS3006

Extralink® Vial

2.5 mL

ESIBIO

GS3009

Extralink® Lite Vial

0.5 mL

ESIBIO

GS3008

Extralink® Lite Vial

2.5 mL

ESIBIO

5050

Fibronectin

1 mg

ESIBIO

GS231

Gelin-S® Thiol-modified Gelatin

1 mL

ESIBIO

GS230

Gelin-S® Thiol-modified Gelatin

5 mL

ESIBIO

GS222

Glycosil® Hyaluronic Acid

1 mL

ESIBIO

GS220

Glycosil® Hyaluronic Acid

5 mL

ESIBIO

GS217

Heprasil® Hyaluronic Acid

1 mL

ESIBIO

GS215

Heprasil® Hyaluronic Acid

5 mL

ESIBIO

5010-D

Nutragen® Bovine Collagen

50 mL

ESIBIO

GS711

PEGDA

1 mL

ESIBIO

GS700

PEGDA

1 g

ESIBIO

GS705

PEGDA

5 g

ESIBIO

GS755

PEGSSDA

0.5 mL

ESIBIO

5020

PEPTITE-2000® RGD Peptide

5 mg

ESIBIO

5005-B

PureCol® Collagen

100 mL

ESIBIO

5007-A

VitroCol® Collagen

20 mL

ESIBIO

5051

Vitronectin

0.1 mg

ESIBIO

EM2203

ExoSense™ CD63 Exosome ELISA Kit

96 assays

ESIBIO

EM2202

VascuNet™ CoCulture Assay Kit

1 Kit

ESIBIO

ST11006

BioLiteTM SSEA-1 (DyLight 488) anti-Human/Mouse Antibody

100 µL

ESIBIO

ST11008

BioLiteTM TRA-1-81 (DyLight 488) anti-Human Antibody

100 µL

ESIBIO

ST11023

Nestin anti-Human Antibody

100 µL

ESIBIO

ST11003

Oct4 anti-Human/Mouse Antibody

100 µL

ESIBIO

ST11001

Sox2 anti-Human/Mouse Antibody

100 µL

ESIBIO

ST11013

SSEA-1 anti-Human/Mouse Antibody

100 µL

ESIBIO

ST11014

SSEA-3 anti-Human/Mouse Antibody

100 µL

ESIBIO

ST11015

SSEA-4 anti-Human Antibody

100 µL

ESIBIO

ST11018

TRA-1-60 (PE) anti-Human Antibody

100 µL

ESIBIO

ST11016

TRA-1-60 anti-Human Antibody

100 µL

ESIBIO

ST11017

TRA-1-81 anti-Human Antibody

100 µL

ESIBIO

ST12007

LIF, Mouse Recombinant

10 µg

ESIBIO

ST12008

LIF, Mouse Recombinant

100 µg

ESIBIO

ES-84

PureStem® 4D20.8, NCr-fac Progenitor

ea

ESIBIO

ES-98

PureStem® E15, Meso-prx/latp Progenitor

ea

ESIBIO

ES-283

PureStem® 7PEND24, NCr-fac Progenitor

ea

ESIBIO

ES-278

PureStem® 7SMOO32, NCr-fac Progenitor

ea

ESIBIO

ES-250

PureStem® SK11, NCr-fac Progenitor

ea

ESIBIO

ES-268

PureStem® MEL2, NCr-fac Progenitor

ea

ESIBIO

ES-256

PureStem® SM30, NCr-fac Progenitor

ea

ESIBIO

ES-335

PureStem® ES-335 Meso-latp Progenitor

ea

ESIBIO

ES-100

PureStem® E72 BETATROPHIN+ Progenitor

ea

ESIBIO

ES-1001

PureStem® ES-1001 GDF11+ Progenitor

ea

ESIBIO

ES-184

PureStem® EN7 Progenitor

ea

ESIBIO

ES-101

PureStem® ES-101 Progenitor

ea

ESIBIO

ES-139

PureStem® ES-139 Progenitor

ea

ESIBIO

ES-154

PureStem® ES-154 Progenitor

ea

ESIBIO

ES-198

PureStem® ES-198 Progenitor

ea

ESIBIO

ES-199

PureStem® ES-199 Progenitor

ea

ESIBIO

ES-209

PureStem® ES-209, Meso-prx/latp Progenitor

ea

ESIBIO

ES-210

PureStem® ES-210, Ecto-ntu Progenitor

ea

ESIBIO

ES-236

PureStem® ES-236 Progenitor

ea

ESIBIO

ES-170

PureStem® E44 Progenitor

ea

ESIBIO

ES-196

PureStem® W10 Progenitor

ea

ESIBIO

ES-194

PureStem® Z11, Meso Progenitor

ea

ESIBIO

EM-1001

PureStem® Progenitor Growth Media, EPM k01

500 mL

ESIBIO

EM-1002

PureStem® Progenitor Growth Media, EPM k02

500 mL

ESIBIO

EM-1003

PureStem® Progenitor Growth Media, EPM k03

500 mL

ESIBIO

EM-1004

PureStem® Progenitor Growth Media, EPM k04

500 mL

ESIBIO

EM-1005

PureStem® Progenitor Growth Media, EPM k05

500 mL

ESIBIO

EM-1006

PureStem® Progenitor Growth Media, EPM k06

500 mL

ESIBIO

EM-1007

PureStem® Progenitor Growth Media, EPM k07

500 mL

ESIBIO

EM-1008

PureStem® Progenitor Growth Media, EPM k08

500 mL

ESIBIO

EM-2002

HyStem®-4D Chondrogenesis Differentiation Kit

1 Kit

ESIBIO

EM-2007

HyStem®-4D Differentiation Kit

1 Kit

ESIBIO

EM-2001

PureStem® Chondrogenesis Differentiation Kit

1 Kit

ESIBIO

EM-2006

PureStem® Choroid Plexus Differentiation Kit

1 Kit

ESIBIO

EM-2003

PureStem® Osteogenesis Differentiation Kit 01

1 Kit

ESIBIO

EM-2004

PureStem® Osteogenesis Differentiation Kit 02

1 Kit

ESIBIO

ST10035

LY411575

5 mg

ESIBIO

ST10008

PD0325901

2 mg

ESIBIO

ST10009

PD0325901

10 mg

ESIBIO

ST10010

PD0325901

415 µL, 10 mM

ESIBIO

ST10034

PD173074

2 mg

ESIBIO

ST10021

RepSox

5 mg

ESIBIO

ST10027

RG108

5 mg

ESIBIO

ST10024

SB203580

2 mg

ESIBIO

ST10012

SB431542

5 mg

ESIBIO

ST10013

SB431542

10 mg

ESIBIO

ST10014

SB431542

1.3 mL, 10 mM

ESIBIO

ST10025

SP600125

5 mg

ESIBIO

ST10015

Thiazovivin

2 mg

ESIBIO

ST10017

XAV939

2 mg

ESIBIO

ST10018

Y27632

2 mg

ESIBIO

ST10019

Y27632

10 mg

ESIBIO

ST10020

Y27632

625 µL, 10 mM

ESIBIO

 

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