FT-ICR
Human kidney tissue was surgically removed during a full
nephrectomy, and remnant tissue was processed for research
purposes by the Cooperative Human Tissue Network (CHTN)
at Vanderbilt University Medical Center. Human biospecimens
were collected in compliance with the CHTN protocols,
institutional IRB policies, and the National Cancer Institute’s
best practices for procurement of remnant surgical research
material. The tissue was flash-frozen over an isopentane-dry
ice slurry and embedded in carboxymethylcellulose. Tissue
sections were cryosectioned with a thickness of 10 μm and
thaw-mounted onto indium tin-oxide-coated glass slides (Delta
Technologies, Loveland, CO, USA). 1,5-Diaminonaphthalene
(DAN) was applied to the tissue surface using a TM Sprayer M3
(HTX Technologies, Chapel Hill, NC, USA). The sample was
imaged (50 μm pitch) directly after matrix application with
a 15T MALDI Fourier-transform ion cyclotron resonance (FT-
ICR) mass spectrometer (Solarix, Bruker Daltonics, Billerica,
MD, USA). Briefly, data were generated from m/z 300-2000
with a 4M file size in negative ion mode.
qTOF
C57BL6/J mice were retro-orbitally infected with S. aureus Newman and sacrificed humanely five days post-infection. Kidneys were flash frozen on an isopentane/dry ice slurry and embedded in 2.6\% carboxymethylcellulose. 5 μm thick sections were collected using a Leica Biosystems CM3050S cryostat and thaw-mounted onto indium tin oxide coated glass slides. Sections were washed with cold 150 mM ammonium formate for 45 seconds for three total washes. 5 mg of 4-(dimethylamino)cinnamic acid matrix was applied using an in-house sublimation device. MALDI IMS data were collected in negative ion mode from m/z 400-2000 using a Bruker MALDI timsTOF fleX platform with a 5 μm step size, 25% laser power, and 25 shots per pixel.
%I 4TU.ResearchData