2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. 如何使用如何使用SpectraMax 微孔读板微孔读板机进机进行快速定量检测行快速定量检测 DNA&RNA 岳岳 兵兵 Product Manager Bing.yuemoldev.com 18501662168 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. 内内容概览容概览 光吸收原理 比色皿、微孔板和超微量 在 SpectraMax 读板机上快速检测DNA & RNA 在SpectraDrop上进行高通量超微量DNA & RNA检测 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. 光吸收检测技术原理光吸收检测技术原理 透射透射率率和和吸光度吸光度 I0 = Ia + It + Ir 入射光强度为 I0，吸收光强度为Ia，透过光强度为It，反射光强度为Ir 因为反射光强度Ir由微孔板材料决定 I0 = Ia + It 朗伯比尔定律 K摩尔吸光系数、a吸光系数,根据C单位决定 透射率透射率 吸收值吸收值 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. 光吸收应用光吸收应用(Absorbance) 直接定量和显色反应直接定量和显色反应 核酸和蛋白质定量核酸和蛋白质定量 酶联免疫检测（酶联免疫检测（ELISAELISA） 酶动力学分析酶动力学分析 细菌内毒素检测细菌内毒素检测 乙酰胆碱脂酶检测乙酰胆碱脂酶检测 细胞活性检测（细胞活性检测（MTTMTT） 环境监测环境监测 等等等等 -0.6-0.4-0.200.20.40.60.811.21.4Series1做标准曲线测定浓度做标准曲线测定浓度 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. DNA、RNA或蛋白浓度检测或蛋白浓度检测 DNA双螺旋中双螺旋中碱基碱基 蛋白质蛋白质酪氨酸酪氨酸、色氨酸、苯丙氨酸成分含量色氨酸、苯丙氨酸成分含量 A = a*c*l a = 物质消光系数物质消光系数 c = 浓度浓度 l= 光光径（径（0.5mm或者或者1mm） SMP软件自动给出软件自动给出 DNA浓度：浓度：OD260=1表示表示 50 g/ml双链双链DNA(dsDNA) DNA纯度：纯度：纯度计算纯度计算(OD260/OD280) DNA纯品纯品: OD260/OD280 = 1.8 决定上决定上 样体积样体积 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. PathCheck光路校正技术（专利）光路校正技术（专利） 比色皿比色皿 光径固定光径固定 通常为通常为1 cm 微孔板微孔板 光径不同，与加液体积有关光径不同，与加液体积有关 1 cm p p 为什么我们需要 PathCheck? 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. 微微孔孔板板 将大量测试管中的实验集中于一个载体上 一块板完成快速大量的检测 Drug Discovery 样品和化学试剂加入孔中 在孔中发生反应 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. 超微量板超微量板 SpectraDrop 2 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. 比色皿适配器比色皿适配器 SpectraCuvette Adapter 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. SpectraMax M5 带带比色皿插槽比色皿插槽 比色皿比色皿 光吸收光吸收 荧光强度荧光强度 化学发光化学发光 6-384孔板孔板 光吸收光吸收 荧光强度荧光强度 荧光偏振荧光偏振 时间分辨荧光时间分辨荧光 化学发光化学发光 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. 微孔板选择微孔板选择 微孔板选择微孔板选择 比色皿选择比色皿选择 普通可见光波段：选择普通全透明板 ，（ Z-dimension 15 mm ） 紫外检测：石英板或紫外透板 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. UV 微孔板选择微孔板选择 微微孔板选孔板选择择 左至右: Quartz, Greiner UV, Costar UV, Polyfiltronics UV and generic polystyrene OD260 values for a water-filled quartz microplate should be 0.030.04 with a standard deviation (SD) 99% aqueous*！）！） SoftMax Pro 软件自动将孔的吸收值换软件自动将孔的吸收值换 算为算为1cm光径的吸收值光径的吸收值 相当于测得了相当于测得了1 cm 光径比色皿的吸光径比色皿的吸 收值收值 可以直接计算样品浓度可以直接计算样品浓度 Absorbance = a*c*p（p=1) 998nm 光吸收跟温度无关光吸收跟温度无关 1 cm p p 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. 专利的光程校正技术专利的光程校正技术 由于加样体积不同造成相同浓度的样品原始OD大小不等 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. 专利的光程校正技术专利的光程校正技术 通过专利的光程校正技术自动 校正之后，OD 都变成相等的了 校正之后的OD 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. SoftMax Pro软件中进行设置软件中进行设置 6/27/2014 Pg. 18 SoftMax Pro 5 SoftMax Pro 6 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. SoftMax Pro Pathcheck Protocol SoftMax Pre-defined Protocol 19 2012 For research use only. Not for use in diagnostic procedures. Trademarks mentioned herein are property of Molecular Devices, LLC or their respective owners. SpectraDrop 超微量检测板 SBS format holder + 2 slide 设计 24 or 64 通量 2uL or 4uL 体积 灵敏度 2ng/uL (dsDNA) 清洗方便 可更换slides 超微量超微