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Matrix-Assisted Laser Desorption/Ionization (MALDI):Matrix molecules absorb laser light, enter an excited state, and collide with sample molecules, facilitating
charge transfer to create ions.
Mass Spectrometric Imaging for biomedical tissue analysisKamila Chughtai and Ron M.A. HeerenChem Rev. Vol.110(5): pp3237–3277, 2010.
MALDI-TOFinstrument
Conventional MALDI plate
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Vacuum sublimation is used to apply an even microscopically thin uniform layer of matrix
compound onto tissue sectionwithout the need for solvents.
Sublimation: the transition of a substance from solid togas phase without an intermediate liquid phase.
MALDI matrices for lipid imaging:
DHB: 2,5-dihydrobenzoic acid (+ve mode)
1,5-diaminonapthalene (-ve mode)
Cryosectioning onto Indium Tin Oxide (ITO) coated glass slides and scanning digital image of slide for
“teaching” FlexControl software on MALDI-TOF.
Minimum 2400dpicryosection image
Cryosectioning 10% gelatinembedded tissue block
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Vacuum sublimation apparatus for matrix application in MALDI Imaging.
Vacuummicro-valve Pirani vacuum
gaugeDigital vacuum
monitor
Vacuumsublimation
chambervacuumexhaust
Cold trap
Matrix deposition by vacuum sublimation.
Matrixcompound
Cold condensor unitof vacuum
sublimation chamber
ITO (indium-tin-oxide coated)slide with cryosections
140°C
0.05 Torr
5‐10°C
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Slides with matrix applied by vacuum sublimation.
Deposition of the matrixcompound is at the molecularlevel because gaseousmolecules recrystallize at therelatively cold surface of thetissue section attached to thecold condenser.
The uniformity of matrixdeposition onto the slideattached to the coldcondenser surface reflectsthe random Brownian motionof the released gaseousmatrix molecules.
Conventional MALDI plate MALDI plate for cryosections
Adapted MALDI plate holds slides forMALDI-IMS.
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Quantitative and SpatialAnalysis of Lipids Involved in
Acute Kidney Injury.
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SWATH (MS/MSALL) on 5600 Triple-TOF Mass Spectrometer
Normalizedkidney weight
Sham surgery or IR
8-10 weeksold C57Bl6/J
NephrectomyMALDI-Imaging MS
on a Bruker-TOF Mass Spectrometer
10% gelatincryosections
SWATH (Sequential Window Acquisition of all Theoretical Mass Spectra
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
Pla
sma
crea
tin
ine
(mg
/dL
)
SHAM controlIR (0.5/6 hrs)
PAS
H&E 100m
Sham IR 0.5/6 hrs IR 0.5/24 hrs
IR (0.5/24 hrs)
*
*
*
0.9
1.0
1.1
1.2
1.3
Plasma creatinine and kidney histology in mice subjected to ischemia/reperfusion (IR) related