MALDI-TOF-MS : imaging X (MRI/MRS) (CT) (PET)
2009 8 8 ( ) 15:39 UTC Imaging mass spectrometry(ims) MALDI (Matrix-Assisted Laser Desorption Ionization ) SIMS (Secondary Ion Mass Spectrometry) DESI (desorption electrospray ionization)
MALDI Matrix Sample LASER Matrix LASER H + Imaging mass spectrometry(ims) Acc Drift + + + + + + 2D Modified from: Lottspeich, Zorbas, eds Bioanalytik, Spektrum Akademischer Verlag, 1998 and M.Stoeckli et al. Analytical BioChemistry 311 (2002)33-39
Principle of MALDI Tissue Imaging 200 µm pixels 2500 5000 7500 10000 m/z 12500 15000 17500 20000 MALDI XY ( ) MALDI-MS
MALDI-Imaging MALDI-TOF MS MALDI-TOF MS MALDI-TOF/TOF MS selected reaction monitoring(srm) MALDI catch freeze OCT Cut cryosection -15-20 C OCT compound
MALDI - Leica OCT milliq 1 70% 1% 2 70% 3 100% Coplin Jar ITO Glass Slides for MALDI Imaging : Part No. 237001 MTP slide-adapter II : Part No. 235380 Glass Slides for MALDI Imaging Part No. 237001 MTP slide-adapter II Part No. 235380 MALDI - OCT compound 10 20 µm 10 12μm (#237001 Glass Slides for MALDI Imaging :BDAL)
MALDI - 5 45 70%EtOH 15 30-1 70%EtOH 15 30-2 96%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): 699-708 Profiling Imaging Droplets Spray coating + + + Acquire mass spectra + + + Protein images m/z 18 388
MALDI - : 1ml 10ml TLC-Sprayer
ImagePrep ImagePrep
Frequency of Occurrence [%] 50 40 30 20 10 0 20µm max 50µm 10 20 30 40 50 60 70 80 90 100 Diameter [µm] 100 µm scattered light intensity time
scattered light intensity thick thin dry wet 0 15 30 Time [min] ImagePrep (50µm crystals) 50 µm Sinapinic acid (20g/l in 50% acetonitrile)
500-800 2500 5000 7500 10000 12500 15000 17500 20000 TIC ( )
ImagePrep 50 µm Sample by courtesy of: Matthias Riemenschneider Neurochemistry and Neurogenetics, Dept. of Psychiatry and Psychotherapy Klinikum rechts der Isar, Munich 50μm MS
1/5 Richard Caprioli, Vanderbilt Univ. TN-USA fleximaging Imaging MS ClinProTools
ROI 1 ROI 3 ROI 2 ROI 4 MALDI / /
MALDI MALDI LC-MS/MS /
MALDI LC-MS/MS /??? MALDI
( ) ( : (*) ( : MALDI
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), 5230-5236 Milk duct ( )
MALDI 1mm MALDI MALDI H&E
Tissue Section Groseclose et al., J. Mass Spectrom. 42, 254-262 (2007)
a.u. 13 12 11 10 9 8 7 6 5 4 3 2 1 a.u. 16 15 14 13 12 11 10 9 8 7 6 5 4 3 2 1 750 1000 1250 1500 1750 2000 2250 2500 2750 3000 3250 3500 3750 750 1000 1250 1500 1750 2000 2250 2500 2750 3000 3250 3500 3750 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 45 40 35 30 25 20 15 10 PV VH y13b7 y11b9 NPVV NP y14b8 y14b6 KNI QD HF y7b14 y17b3 y10b10 FKN 344.628 y8b13 b 3 NPV 271.702 y14b7 y 2 229.684 H b 2 372.656 y 3 174.773 y 1 73.865 a 1 R P T 559.678 a 5 DEN y16b5 FK y8b12 471.700 y 4 473.611 b 4 FF y9b11 587.640 FFK b 5 FFKN PVVHFF y9b12 HFFKNI NPVVHFFKN y9b13 y13b11 y10b14 y14b13 ENP VHF QDEN HFFKN y15b6 y11b10 VHFFKN HFFKNIVT y17b5 y10b13 y11b13 y10b16 698.806 1667.452 HFF DENPV VVHFFK y 6 VVHFFKN y 14 y10b11 y16b7 y12b12 y12b13 684.694 1641.939 854.906 b 6 1258.131 a 14 a 8 y 10 1019.887 656.727 1553.384 826.888 b 9 1166.872 NPVVHFFK a 6 y 13 y 7 b 10 y14b12 991.911 584.743 783.757 a 9 1556.282 y 5 1138.985 VHFFKNI b 13 b 7 a 10 y11b14 973.971 755.813 y 8 1121.061 1443.138 a 7 y 9 b 12 1357.223 882.855 y 11 b 8 FFKNI y9b14 1314.424 b 11 1796.514 y 15 2140.902 y 18 2094.526 a 18 2040.678 y 17 1967.675 b 17 1911.835 y 16 5 I 200 400 600 800 1000 1200 1400 1600 1800 2000 m/z
ImageID TM MALDI LC- MALDI FFPE
MALDI Imaging 20μm
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):2048-54
1 2 3 MALDI SRM (Selected Reaction Monitoring) ImagingMS
MALDI SRM (Selected Reaction Monitoring) interest peaks MS MS/MS m/z MS/MS MS/MS1 MS/MS2 SRM FAST-SRM Imaging
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 :
(OLZ) SRM-IMS(m/z 313->256) control 8mg/kg p.o. 2h 20mg/kg p.o. 2h 100mg/kg p.o. 2h
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
Intens. [a.u.] Intens. [a.u.] Intens. [a.u.] Intens. [a.u.] 5000 4000 3000 2000 1000 0 4000 3000 2000 1000 0 8000 6000 4000 2000 0 x10 4 0.8 0.6 0.4 0.2 BS01_NaTFA 0:D13 MS Mn; 1849.7 Mw; 1919.7 BS02_NaTFA 0:F14 MS Mn; 1969.9 Mw; 2038.1 BS04_NaTFA 0:I14 MS Mn; 2065.0 Mw; 2124.3 BS06_NaTFA 0:L14 MS Mn; 2078.0 Mw; 2133.9 1000 1500 2000 2500 3000 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.] 1000 750 500 250 0 x10 4 0.8 0.6 0.4 0.2 0.0 8000 6000 4000 2000 0 4000 2000 BS01Disk_NaTFA 0:E16 MS Mn; 1748.2 Mw; 1825.0 BS02Disk_NaTFA 1:C14 MS Mn; 1988.1 Mw; 2080.4 BS04Disk_NaTFA 0:C15 MS Mn; 2069.8 Mw; 2150.9 BS06Disk_NaTFA 1:C15 MS Mn; 2086.8 Mw; 2168.7 1000 1500 2000 2500 3000 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 60 1500 B (%) 40 20 0 No.1 No.2 No.3 No.4 1000 500 0 No.1 No.2 No.3 No.4 No.1 No.2 No.3 No.2 2 No.4 No.3 2
2 Imaging MS A B 2mm m/z 1375.1 m/z 1383.1 m/z 1673.5 C (Mono) D (Bis) E (Mono) F m/z 1680.5 (Bis) 2mm 2mm 2mm 2mm A; / B; 1500 C; m/z 1375.1 D; m/z 1383.1 E; m/z 1673.5 F; m/z 1680.5 www.bruker.com