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. 2008 Dec 1;278(2-3):143-149.
doi: 10.1016/j.ijms.2008.04.005.

A Minimalist Approach to MALDI Imaging of Glycerophospholipids and Sphingolipids in Rat Brain Sections

Hay-Yan J Wang  1 , Shelley N Jackson, Jeremy Post, Amina S Woods
Affiliations

A Minimalist Approach to MALDI Imaging of Glycerophospholipids and Sphingolipids in Rat Brain Sections

Hay-Yan J Wang et al. Int J Mass Spectrom. .

Abstract

Matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) is a powerful tool that has allowed researchers to directly probe tissue molecular structure and drug content with minimal manipulations, while maintaining anatomical integrity. In the present work glycerophospholipids and sphingolipids images were acquired from 16 µm thick coronal rat brain sections using MALDI-MS. Images of phosphatidylinositol 38:4 (PI 38:4), suifatide 24:1 (ST 24:1), and hydroxyl sulfatide 24:1 (ST 24:1 (OH)) were acquired in negative ion mode, while the images of phosphatidylcholine 34:1 (PC 34:1), potassiated phosphatidylcholines 32:0 (PC32:0 + K(+)) and 36:1 (PC 36:1 +K(+)) were acquired in positive ion mode. The images of PI 38:4 and PC 36:1+K(+) show the preferential distribution of these two lipids in gray matter; and the images of two sulfatides and PC 32:0+K(+) show their preferential distribution in white matter. In addition, the gray cortical band and its adjacent anatomical structures were also identified by contrasting their lipid makeup. The resulting images were compared to lipid images acquired by secondary ion mass spectrometry (SIMS). The suitability of TLC sprayers, Collison Nebulizer, and artistic airbrush were also evaluated as means for matrix deposition.

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Figures

Figure 1
Figure 1
Surface light microscope pictures of a target plate sprayed with DHB. Left panel: Sprayed with TLC sprayer. Middle panel: Sprayed with Collison Nebulizer over a tissue section. Note the section was not covered by the matrix. Right panel: Sprayed with air brush. Note the homogeneous deposition of matrix over the target surface as compared to the TLC deposition method. Scale bar: 5 mm.
Figure 2
Figure 2
Light microscope photograph of a rat brain section. CC: corpus callosum; CTX: cortex; FMJ: forceps major of corpus callosum; HIP: hippocampus; MRN: midline raphe nuclei; PAG: periaquiductal gray; RF: rhinal fissure; SNR: substantia nigra par reticulata; VTA: ventral tegmental area. *: Figure 4A region; #: Figure 4B region; @: Figure 6A region; %: Figure 6B region. Scale bar: 5 mm.
Figure 3
Figure 3
images of three phospholipids acquired in negative ion mode: left panel: phosphatidylinositol 38:4, m/z 885; middle panel: sulfatide 24:1, m/z 888; right panel: hydroxy sulfatide 24:1, m/z 906. The color scale to the right of each panel represents the raw ion count. Scale bar: 5 mm.
Figure 4
Figure 4
Figure 4A: Negative ion mode MALDI TOF spectrum acquired from gray matter region indicated in Figure 1. Figure 4B: Negative ion mode MALDI TOF spectrum acquired from white matter region indicated in Figure 1.
Figure 4
Figure 4
Figure 4A: Negative ion mode MALDI TOF spectrum acquired from gray matter region indicated in Figure 1. Figure 4B: Negative ion mode MALDI TOF spectrum acquired from white matter region indicated in Figure 1.
Figure 5
Figure 5
images of three common lipids acquired in positive ion mode: left panel: phosphatidylcholine 34:1, m/z 760; middle panel: potassiated phosphatidylcholine 32:0, m/z 772; right panel: phosphatidylcholine 36:1, m/z 826. The color scale to the right of each panel represents the raw ion count. Scale bar: 5 mm.
Figure 6
Figure 6
Figure 6A: Positive ion mode MALDI-TOF spectrum acquired from the gray matter region indicated in Figure 1. Figure 6B: Positive ion mode MALDI-TOF spectrum acquired from the white matter region indicated in Figure 1.
Figure 6
Figure 6
Figure 6A: Positive ion mode MALDI-TOF spectrum acquired from the gray matter region indicated in Figure 1. Figure 6B: Positive ion mode MALDI-TOF spectrum acquired from the white matter region indicated in Figure 1.
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