Supplementary Information
Fully Automated Laser Ablation Liquid Capture Surface
Analysis using NanoElectrospray Ionization Mass Spectrometry
Matthias Lorenz, Olga S. Ovchinnikova, and Gary J. Van Berkel*
Organic and Biological Mass Spectrometry Group, Chemical Sciences Division,
Oak Ridge National Laboratory, Oak Ridge, TN 37831-6131, USA
*Corresponding Author
Phone: 865-574-1922
FAX: 865-574-4961
E-mail: vanberkelgj@ornl.gov
This manuscript has been authored by a contractor of the U.S. Government under
contract DE-AC05-00OR22725. Accordingly, the U. S. Government retains a paid-up,
nonexclusive, irrevocable, worldwide license to publish or reproduce the published form of this
contribution, prepare derivative works, distribute copies to the public, and perform publicly and
display publicly, or allow others to do so, for U.S. Government purposes.
Figure S1. Absolute capture efficiency. Comparison of the integrated signal intensities of the
cationic dye basic blue 7 (SRM m/z 478  434, CE = 65 eV), generated from LA/LCSA of a
spot (120 µm x 139 µm) and a 60 s direct liquid extraction analysis (LESA®) of a laser etched
square (122 µm x 122 µm) of a thin-film layer of blue sharpie marker containing basic blue 7 on
a glass microscope slide. Solvent: 80/20/0.1 (v/v/v) methanol/water/formic acid. Sampling
conditions LESA®: 1 µL droplet, 2 µL total solvent volume, 3 µL re-aspirated, 60 s sampling.
Sampling conditions LA/LCSA: 0.4 µL droplet, 1.2 mm probe-to-sample distance, 2 µL total
solvent volume, 3 µL re-aspirated. Laser: 355 nm, Nd:YAG, 1 kHz, (120 µm x 139 µm) spot
Signal per Surface Area (a.u./mm2)
Signal per Surface Area (a.u. / mm2)
size, 9 µJ pulse energy on the surface, 15 s ablation.
4
1.5x10
4
1.0x10
3
5.0x10
0.0
LESA
LA/LCSA
LESA
Laser
LESA
Sampling
Mode
Sampling Mode
Figure S2. Analyte dissolution and mixing. Extracted ion chronograms for rhodamine B (SRM
m/z 443  399, CE = 65 eV black trace) and internal standard 0.1 µM crystal violet (SRM m/z
372  356, CE = 65 eV, blue trace) generated from a 1 s laser ablation of a thin-film layer of
pink sharpie marker containing rhodamine B on a glass microscope slide followed by a 1 µL, 2
µL, 3 µL and 4 µL re-aspiration of the capture solvent back into the pipette tip. All signal
intensities were normalized to the maximum recorded; the signal intensity for crystal violet (blue
line) was multipled by 10. Solvent: 80/20/0.1 (v/v/v) methanol/water/formic acid. Sampling
conditions: 0.2 µL droplet, 1.2 mm probe-to-sample distance, 7.5 µL total solvent volume.
Laser: 355 nm, Nd:YAG, 1 kHz, 155 µm spot size, 9 µJ pulse energy on the surface, 1 s ablation.
Rel. Int. (%)
100
1 µL re-aspiration
volume
2 µL
3 µL
4 µL
50
0
0
5
0
5
time (min)
0
5
0
5
10
Figure S3. Full scan product ion spectrum (m/z 100-600) of m/z 595 (CID with 40 eV) obtained
from laser drilling of the brown pigmented tissue material of an Alstroemeria Yellow King
flower leaf followed by LESA® analysis directly on the ablation spot (i.e., LA/LESA®). Solvent:
80/20/0.1 (v/v/v) methanol/water/formic acid. Sampling conditions LESA®: 1 µL droplet, 10
µL total solvent volume, 3 µL re-aspirated, 60 s sampling. Laser: 355 nm, Nd:YAG, 1 kHz, 120
µm spot size, 9 µJ pulse energy on the surface, 15 s exposure.
Rel. Int. (%)
100
287
308 Da
50
0
100
200
300
400
m/z
500
600