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1 MALDI-TOF-MS : imaging X (MRI/MRS) (CT) (PET)
2 ( ) 15:39 UTC Imaging mass spectrometry(ims) MALDI (Matrix-Assisted Laser Desorption Ionization ) SIMS (Secondary Ion Mass Spectrometry) DESI (desorption electrospray ionization)
3 MALDI Matrix Sample LASER Matrix LASER H + Imaging mass spectrometry(ims) Acc Drift D Modified from: Lottspeich, Zorbas, eds Bioanalytik, Spektrum Akademischer Verlag, 1998 and M.Stoeckli et al. Analytical BioChemistry 311 (2002)33-39
4 Principle of MALDI Tissue Imaging 200 µm pixels m/z MALDI XY ( ) MALDI-MS
5 MALDI-Imaging MALDI-TOF MS MALDI-TOF MS MALDI-TOF/TOF MS selected reaction monitoring(srm) MALDI catch freeze OCT Cut cryosection C OCT compound
6 MALDI - Leica OCT milliq 1 70% 1% 2 70% 3 100% Coplin Jar ITO Glass Slides for MALDI Imaging : Part No MTP slide-adapter II : Part No Glass Slides for MALDI Imaging Part No MTP slide-adapter II Part No MALDI - OCT compound µm 10 12μm (# Glass Slides for MALDI Imaging :BDAL)
7 MALDI %EtOH %EtOH %EtOH pure EtOH ) 15 Neuropeptide Direct tissue analysis using matrix-assisted laser desorption/ionization mass spectrometry: practical aspects of sample preparation Schwartz SA, Reyzer ML, Caprioli RM, J. Mass Spectrom. (2003); 38(7): Profiling Imaging Droplets Spray coating Acquire mass spectra Protein images m/z
8 MALDI - : 1ml 10ml TLC-Sprayer
9 ImagePrep ImagePrep
![Diameter [µm] 100 µm](/docs-images/77/74858474/images/10-2.jpg "scattered light")
10 Frequency of Occurrence [%] µm max 50µm Diameter [µm] 100 µm scattered light intensity time
11 scattered light intensity thick thin dry wet Time [min] ImagePrep (50µm crystals) 50 µm Sinapinic acid (20g/l in 50% acetonitrile)
12 TIC ( )
13 ImagePrep 50 µm Sample by courtesy of: Matthias Riemenschneider Neurochemistry and Neurogenetics, Dept. of Psychiatry and Psychotherapy Klinikum rechts der Isar, Munich 50μm MS
14 1/5 Richard Caprioli, Vanderbilt Univ. TN-USA fleximaging Imaging MS ClinProTools
15 ROI 1 ROI 3 ROI 2 ROI 4 MALDI / /
16 MALDI MALDI LC-MS/MS /
17 MALDI LC-MS/MS /??? MALDI
18 ( ) ( : (*) ( : MALDI
19 Connective Tissue Tumor Clone 1 Tumor Clone 2 Inflammation Deininger, S., Ebert, M., Fütterer A., Gerhard M., Röcken, C. (2008) J. Proteome Res. 7(12), Milk duct ( )
20 MALDI 1mm MALDI MALDI H&E
21 Tissue Section Groseclose et al., J. Mass Spectrom. 42, (2007)
22 a.u a.u m/z m/z vs. m/z 2141 MBP TOF/TOF LIFT MS/MS MASCOT m/z 2141 Myelin Basic Protein MASCOT Score: 123 Abs. Int. * 1000 a T P V V H F b D E N P V V H F F K N y I N R P T V K F F H V N E D Q T PV VH y13b7 y11b9 NPVV NP y14b8 y14b6 KNI QD HF y7b14 y17b3 y10b10 FKN y8b13 b 3 NPV y14b7 y H b y y a 1 R P T a 5 DEN y16b5 FK y8b y b 4 FF y9b FFK b 5 FFKN PVVHFF y9b12 HFFKNI NPVVHFFKN y9b13 y13b11 y10b14 y14b13 ENP VHF QDEN HFFKN y15b6 y11b10 VHFFKN HFFKNIVT y17b5 y10b13 y11b13 y10b HFF DENPV VVHFFK y 6 VVHFFKN y 14 y10b11 y16b7 y12b12 y12b b a 14 a 8 y b NPVVHFFK a 6 y 13 y 7 b 10 y14b a y VHFFKNI b 13 b 7 a 10 y11b y a 7 y 9 b y 11 b 8 FFKNI y9b b y y a y b y 16 5 I m/z
23 ImageID TM MALDI LC- MALDI FFPE
24 MALDI Imaging 20μm
25 Lipid Imaging at 10μm Smartbeam laser: Industry-leading performance in MALDI imaging at 2 khz Different lipid species in mouse brain measured at a spatial resolution of 10 µm on an ultraflextreme. from: Chaurand P et al. Sublimation of new matrix candidates for high spatial resolution imaging mass spectrometry of lipids: enhanced information in both positive and negative polarities after 1,5-diaminonapthalene deposition. Anal Chem. 2012;84(4):
26 1 2 3 MALDI SRM (Selected Reaction Monitoring) ImagingMS
27 MALDI SRM (Selected Reaction Monitoring) interest peaks MS MS/MS m/z MS/MS MS/MS1 MS/MS2 SRM FAST-SRM Imaging
28 FAST-SRM (OLZ) MS-IMS OLZ(m/z 313) 100mg/kg p.o. 2h 100mg/kg p.o. 6h 100mg/kg p.o. 2h 100mg/kg p.o. 6h Control 20mg/kg p.o. 2h 20mg/kg p.o. 2h N-desmethyl metabolite (m/z 299) Control 20mg/kg p.o. 2h 20mg/kg p.o. 6h 100mg/kg p.o. 2h 100mg/kg p.o. 6h 100mg/kg p.o. 2h 100mg/kg p.o. 6h Control 20mg/kg p.o. 2h 20mg/kg p.o. 6h Control 20mg/kg p.o. 2h 20mg/kg p.o. 6h :
29 (OLZ) SRM-IMS(m/z 313->256) control 8mg/kg p.o. 2h 20mg/kg p.o. 2h 100mg/kg p.o. 2h
30 2 HOCH 2 CH HO CH 2 O CH 2 CF 2 OCF 2 m OCF 2 CF 2 n OCF 2 CH 2 O CH 2 CH 2 OH CH OH HOCH 2 CH HO CH 2 O CH 2 CF 2 OCF 2 m OCF 2 CF 2 n OCF 2 CH 2 O CH 2 CH 2 CH OH O CH 2 CH 2 OH CH OH SEM 60
31 Intens. [a.u.] Intens. [a.u.] Intens. [a.u.] Intens. [a.u.] x BS01_NaTFA 0:D13 MS Mn; Mw; BS02_NaTFA 0:F14 MS Mn; Mw; BS04_NaTFA 0:I14 MS Mn; Mw; BS06_NaTFA 0:L14 MS Mn; Mw; m/z sample 1 PFPE A:B mix ratio 100 : 0 sample 2 PFPE A:B mix ratio 98 : 2 sample 3 PFPE A:B mix ratio 79 : 21 sample 4 PFPE A:B mix ratio 0 : 100 Intens. [a.u.] Intens. [a.u.] Intens. [a.u.] Intens. [a.u.] x BS01Disk_NaTFA 0:E16 MS Mn; Mw; BS02Disk_NaTFA 1:C14 MS Mn; Mw; BS04Disk_NaTFA 0:C15 MS Mn; Mw; BS06Disk_NaTFA 1:C15 MS Mn; Mw; m/z 61 1 Endgroup A HOCH 2 CH HO CH 2 O CH 2 CF 2 OCF 2 m OCF 2 CF 2 n OCF 2 CH 2 O CH 2 CH 2 OH CH OH Endgroup B HOCH 2 CH HO CH 2 O CH 2 CF 2 OCF 2 m OCF 2 CF 2 n OCF 2 CH 2 O CH 2 CH 2 CH OH O CH 2 CH 2 OH CH OH B (%) No.1 No.2 No.3 No No.1 No.2 No.3 No.4 No.1 No.2 No.3 No.2 2 No.4 No.3 2
32 2 Imaging MS A B 2mm m/z m/z m/z C (Mono) D (Bis) E (Mono) F m/z (Bis) 2mm 2mm 2mm 2mm A; / B; 1500 C; m/z D; m/z E; m/z F; m/z
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質量分析セミナー autoflex III LRF 韮澤崇 Bruker Daltonics K.K. MALDI Matrix Assisted Laser Desorption / Ionization マトリックス支援レーザー脱離イオン化 2 MALDI の基本原理 Matrix と Sample の混合結晶 LASER による Matrix の励起 脱離およびイオン化 ( プロトン化 ) LASER
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Mass Spectroscopy(MS) EI electron impact, electron impact ionization, electron bombardment ionization ( ) IUPAC electron ionization ( ) ( 31 (1997)) MS m/z( / ) 1 C 12.011 C 98.9% 13 C 1.1% m/z m z m/z
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2017 5 26 350 Restek time time time increasing time μ = = k = t r t o t o = t r ' t o van Deemter Equation H A B H = height equivalent theoretical plate (HETP) A = Eddy diffusion term B = Molecular diffusion
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