The molecular scanner in microscope mode.
The combination of microscope mode matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) with protein identification methodology: the molecular scanner, was explored. The molecular scanner approach provides improvement of sensitivity of detection and identification of high-mass proteins in microscope mode IMS. The methodology was tested on protein distributions obtained after separation by sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE). High-quality, high-spatial-resolution ion images were recorded on a TRIFT-II ion microscope after gold coating of the MALDI sample preparation on the poly(vinylidenedifluoride) capture membranes. The sensitivity of the combined method is estimated to be 5 pmol. The minimum amount of sample consumed, needed for identification, was estimated to be better than 100 fmol. Software tools were developed to analyze the spectral data and to generate broad mass range and single molecular component microscope mode ion images and single mass-to-charge ratio microprobe mode images.
Luxembourg
Stefan L
SL
FOM Institute for Atomic and Molecular Physics, Kruislaan 407, 1098 SJ Amsterdam, The Netherlands.
Vaezaddeh
Ali R
AR
Amstalden
Erika R
ER
Zimmermann-Ivol
Catherine G
CG
Hochstrasser
Denis F
DF
Heeren
Ron M A
RM
D000818:Animals; D001709:Biotechnology; D002417:Cattle; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D040901:Proteomics / Q000379:methods; D011817:Rabbits; D012542:Scattering, Radiation; D012680:Sensitivity and Specificity; D020539:Sequence Analysis, Protein / Q000295:instrumentation / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation / Q000379:methods*; D018629:Spectrometry, Mass, Secondary Ion / Q000379:methods*
D011506:Proteins