MALDITOFTOFMS在蛋白质组研究中的应用.pptVIP

TOF/TOF 质谱（Tandem Spectrometer with CID） Ion Source #2 Ion Source #1 Precursor Ion Selector (TIS) Collision Cell Mass Analyzer #2 Detector Mass Analyzer #1 Generates and Accelerates Ions TOF 1 Separates Ions Induces CID Fragmentation of Precursor Re-Accelerates Fragments TOF 2 with Mirror Separates Fragments Selects Ions . * . 如何实现MS/MS分析？ 凭借时间离子选择装置（Timed-Ion-Selector，TIS)选择母离子，在CID中与一定压力的惰性气体分子发生碰撞得到碎片离子，然后进行TOF分离和检测。 TIS具有两种功能：一是可以抑制来自基质和样品中不感兴趣的低质量的离子；二是为MS/MS分析选择母离子。 操作步骤：依据选择离子达到的时间适时的施加电压（关门）偏转离子的飞行轨道或接地（开门）使离子通过。 . * . Timed-Ion-Selector Voltage Detector Detector Output Time Intensity + + + Mass 50 Mass 100 Mass 500 Mass 50 Accel Drift + + + Time = 0 - + . * . Timed Ion Selector Voltage Detector Mass 50 Mass 100 Mass 500 Detector Output Mass 50 Time Intensity Accel Drift Time = A - + + + + Mass 50 Mass 100 Mass 500 Mass 50 . * . Timed Ion Selector Time = B Voltage Detector Detector Output Time Intensity Accel Drift + + + Mass 50 Mass 100 Mass 500 Mass 50 . * . Timed Ion Selector Voltage Detector Detector Output Time Intensity Accel Drift Time = C - + C + + + Mass 50 Mass 100 Mass 500 Mass 50 . * . Timed Ion Selector Voltage Detector Detector Output Time Intensity Accel Drift Time = D + - C + + + Mass 50 Mass 100 Mass 500 Mass 50 . * . Pulsed laser 3. Ions are accelerated by an electric field to the same kinetic energy; they separate according to mass as they drift thru the field-free region of the flight tube Flight tube 1. Sample + matrix dried on target plate High vacuum Tim e High voltage 2. Sample is irradiated with laser, clock starts to measure time-of-flight 20 - 25 kV 5. A data system controls instrument parameters, acquires signal vs. time, and processes the data 4. Ions strike the detector at different times depending on the mass/charge ratio 一般MALDI质谱的分析步骤 . * . 离子源 基质辅助激光解吸电离(MALDI) 基本原理：将分析物分散在基质(Matrix)分子中并形成共结晶, 当用激光(337 nm的氮激光或355 nm的固体激光器)照射晶体时, 样品分子获得激光能量解吸附，基质－样品之间发生电荷转移使样品分子电离。 离子源特点 1、