Selected MALDI Imaging Publications - Bruker Daltonics

Innovation with Integrity 

MALDI Imaging in the 

Scientific Literature 

Selected Publications 

Mass Spectrometry

Figure 1: Drug Imaging / Histopharmacology: 

Correlation of 

histology and ion image in dog 

liver sections. The ion image (50 

µm spatial resolution) shows the 

lapatinib metabolite M10, localized 

only in the regions associated with 

the inflammation. Adapted from: 

Castellino et al. (2011) Bioanalysis 

3(21),2427-41 with kind permission 

of the authors. 

Selected MALDI Imaging Publications 

Reviews 

Selected review articles on different MALDI 

Imaging application areas 

Castellino S, Groseclose MR, Wagner D (2011( 

MALDI imaging mass spectrometry: bridging 

biology and chemistry in drug development. 

Bioanalysis. 3(21):2427-41 

This review explains how the combination 

of MALDI imaging with histology can solve 

problems in drug development. The review 

is illustrated with real life example from 

ClaxoSmithKline‘s DMPK unit, the data are 

acquired on a Solarix instrument. Highlights of 

the presented data include a Lapatinib metabolite 

that is only found in inflamed lesions in 

the liver, and another metabolite that is highly 

enriched in small bile ducts. 

Schwamborn K, Caprioli RM (2010) 

Molecular imaging by mass spectrometry - 

looking beyond classical histology. 

Nat Rev Cancer. 10(9):639-46 

A comprehensive review on imaging mass 

spectrometry in clinical cancer research. 

Walch A, Rauser S, Deininger SO, Hofler H 

(2008) 

MALDI Imaging mass spectrometry for direct 

tissue analysis: a new frontier for molecular 

histology. 

Histochem Cell Biol. 130(3):421-34 

A general review on imaging mass spectrometry 

with a detailed description on the importance 

of the combining histological information 

with MALDI Imaging. 

H&E stain 

Serial tissue sections 

Rauser S, Deininger SO, Suckau D, Hofler H, 

Walch A (2010) 

Approaching MALDI molecular imaging for 

clinical proteomic research: current state and 

fields of application. 

Expert Rev Proteomics. 7(6):927-41 

This review focuses on the application of 

MALDI Imaging for clinical proteomic research. 

It features an extensive discussion and explanation 

of relevant clinical questions where MALDI 

Imaging can be applied. 

Bonnel D, Legouffe R, Willand N, Baulard A, 

Hamm G, Deprez B, Stauber J (2011) 

MALDI Imaging techniques dedicated to drugdistribution 

studies. 

Bioanalysis. 3(12):1399-406 

This review describes the use of MALDI 

Imaging for drug-distribution studies. It also 

features an extensive discussion of Brukers 

innovative FAST-SRM mode for drug imaging 

on MALDI-TOF instruments. 

Grassl J, Taylor NL, Millar AH (2011) 

Matrix-assisted laser desorption/ionisation 

mass spectrometry imaging and its development 

for plant protein imaging. 

Plant Methods. 7(1):21 

This review describes the use of MALDI 

Imaging for plant research. 

Ion images

Clinical Research 

MALDI Imaging is perfectly suited for clinical 

biomarker studies. This sections contains 

selected papers that have used MALDI 

Imaging to study human patient samples 

Meding S, Nitsche U, Balluff B, Elsner M, 

Rauser S, Schone C, Nipp M, Maak M, Feith M, 

Ebert MP, Friess H, Langer R, Hofler H, Zitzelsberger 

H, Rosenberg R, Walch A (2012) 

Tumor Classification of Six Common Cancer 

Types Based on Proteomic Profiling by MALDI 

Imaging. 

J Proteome Res. 11(3):1996-2003 

The carcinoma of unknown primary (CUP) is a 

challenge in clinical oncology. MALDI imaging 

can classify tissue based on protein profiles. In 

this article, the authors describe how molecular 

classifiers allowed to differentiate different 

types of tumor. Interestingly, they were able to 

classify liver metastases from a known origin 

(colon) correctly. This shows that, in principle, a 

classification of metastases of unknown origin 

might be possible. 

Balluff B, Rauser S, Meding S, Elsner M, 

Schone C, Feuchtinger A, Schuhmacher C, 

Novotny A, Jutting U, Maccarrone G, Sarioglu 

H, Ueffing M, Braselmann H, Zitzelsberger 

H, Schmid RM, Hofler H, Ebert MP, Walch A 

(2011) 

MALDI Imaging Identifies Prognostic Seven- 

Protein Signature of Novel Tissue Markers in 

Intestinal-Type Gastric Cancer. 

Am J Pathol. 179(6):2720-9 

A comprehensive MALDI imaging study of 

gastric cancer: A seven protein signature was 

found to correlate with survival in a discovery 

cohort (n= 63). Three of the proteins were 

identified and the findings from the discovery 

cohort were subsequently validated on a larger 

independet cohort by immunohistochemistry. 

Hardesty WM, Kelley MC, Mi D, Low RL, 

Caprioli RM (2011) 

Protein signatures for survival and recurrence in 

metastatic melanoma. 

J Proteomics. 74(7):1002-14 

A study that utilised MALDI Imaging for survival 

and reccurence in metastatic melanoma. 

Twelve proteins could be associated with 

survival and three proteins were found to be 

associated with reccurence. 

Le Faouder J, Laouirem S, Chapelle M, 

Albuquerque M, Belghiti J, Degos F, Paradis V, 

Camadro JM, Bedossa P (2011) 

Imaging mass spectrometry provides fingerprints 

for distinguishing hepatocellular carcinoma 

from cirrhosis. 

J Proteome Res. 10(8):3755-65 

The differentiation of liver carcinoma and its 

predecessor, liver cirrhosis, was analyzed by 

MALDI Imaging. Among other mass signals, 

Ubiquitin was found to be up-regulated in the 

tumors, as was confirmed by immunohistochemistry. 

This paper also uses a supervised 

classification to differentiate cancerous from 

cirrhotic regions and shows how molecular 

images can be constructed from classification 

results („Classimaging“). 

Han EC, Lee YS, Liao WS, Liu YC, Liao HY, 

Jeng LB (2011) 

Direct tissue analysis by MALDI-TOF mass 

spectrometry in human hepatocellulat carcinoma. 

Clin Chim Acta. 412(3-4):230-9 

At present, prognosis of hepatocellular carcinoma 

(HCC) relies primarily on histopathological 

imaging applications after surgical removal. 

In this study, MALDI Imaging was utilised for 

classification of various tumour to supplement 

current histophatological analysis in tumour 

margins. A progressive change of fourteen 

peaks was observed in a distance-dependent 

manner from non-tumour to tumour regions of 

the junction section of HCC. 

Zaima N, Sasaki T, Tanaka H, Cheng XW, 

Onoue K, Hayasaka T, Goto-Inoue N, Enomoto 

H, Unno N, Kuzuya M, Setou M (2011) 

Imaging mass spectrometry-based histopathologic 

examination of atherosclerotic lesions. 

Atherosclerosis. 217(2):427-32 

This article describes the investigation of 

artherosclerotic lesions of ApoE deficient mice 

and humans with peripheral artery occlusive 

disease using IMS-based histopathologic 

examination (IbHE). Several lipids such as 

cholesterol linoleate (CE 18:2) and cholesterol 

oleate (CE 18:1) were specifically found in the 

lipid rich regions of both human and mouse, 

whilst triacylglycerol was localised in the 

lipid-rich regions of human samples. Other 

molecules were found to be localised in the 

calcified regions.

Schwamborn K, Krieg RC, Jirak P, Ott G, 

Knuchel R, Rosenwald A, Wellmann A (2010) 

Application of MALDI Imaging for the diagnosis 

of classical Hodgkin lymphoma. 

J Cancer Res Clin Oncol. 136(11):1651-5 

In this study, Hodgkin lymphoma was investigated 

by MALDI Imaging. the differentially 

expressed protein profiles allowed the differentiation 

between classical Hodgkin lymphoma 

and lymphadenitis with a sensitivity and specificity 

of 83.92 and 89.37%, respectively. 

Tanaka H, Zaima N, Yamamoto N, Sagara D, 

Suzuki M, Nishiyama M, Mano Y, Sano M, 

Hayasaka T, Goto-Inoue N, Sasaki T, Konno H, 

Unno N, Setou M (2010) 

Imaging mass spectrometry reveals unique 

lipid distribution in primary varicose veins. 

Eur J Vasc Endovasc Surg. 40(5):657-63 

Various lipid molecules have been measured 

in varicose and normal human veins. The 

accumulation of various specific lipids (identified 

using a combination of imaging mass 

spectrometry and MS/MS) in varicose veins 

suggested that inflammation with abnormal 

lipid metabolism is involved in the devlopment 

of varicose veins. 

Oppenheimer SR, Mi D, Sanders ME, 


Molecular analysis of tumor margins by MALDI 

mass spectrometry in renal carcinoma. 


In surgery it is important to evaluate that the 

tumor margin has been completely resected. 

The tumor margin is usually evauated by histology. 

The MALDI Imaging analysis of tumor 

margins in renal carcinoma in this study revelaed 

that proteins that characteristically upregulated 

in the tumour were also up-regulated 

in an area adjacent to the tumor, indicating a 

molecular tumor margin that is different from 

the histological tumor margin. 

Balluff B, Elsner M, Kowarsch A, Rauser S, 

Meding S, Schuhmacher C, Feith M, Herrmann 

K, Rocken C, Schmid RM, Hofler H, Walch A, 

Ebert MP (2010) 

Classification of HER2/neu status in gastric 

cancer using a breast-cancer derived proteome 

classifier. 


A paper that follows on an earlier breast cancer 

study (Rauser et al. J. Proteome Res. , 2010). 

It showed that a protein profile based classifier, 

trained to classify Her2 status in breast cancer, 

was also able to classify Her 2 status in an 

independent gastric cancer cohort, providing 

strong evidence of the robustness of MALDI 

Imaging based tissue classifications. 

Willems SM, van Remoortere A, van Zeijl R, 

Deelder AM, McDonnell LA, Hogendoorn PC 

(2010) 

Imaging mass spectrometry of myxoid sarcomas 

identifies proteins and lipids specific to 

tumour type and grade, and reveals biochemical 

intratumour heterogeneity. 

J Pathol. 222(4):400-9 

An article that investigates the expression 

of proteins and lipids in myxoid sarcomas. 

Peptide, protein and lipid profiles have been 

correlated with tumor grades and intra-tumor 

heterogeneity has been investigated by hiearchical 

clustering of imaging data. In particular, 

lipid changes observed in myxoid liposarcomas 

could be related to pathways known to be 

affected during tumor development. 

Rauser S, Marquardt C, Balluff B, Deininger 

SO, Albers C, Belau E, Hartmer R, Suckau D, 

Specht K, Ebert MP, Schmitt M, Aubele M, 

Hofler H, Walch A (2010) 

Classification of HER2 receptor status in breast 

cancer tissues by MALDI Imaging mass spectrometry. 


In this article, MALDI Imaging has been used 

to classify HER2 receptor status in human 

breast cancer specimens. Protein profiles 

correlated strongly with HER2 expression. A 

protein signal with a particularly high correlation 

with HER2 expression has been identified as 

cysteine rich intestinal protein 1, a protein that 

was previously not known to be associated 

with HER2 expression. 

Cazares LH, Troyer D, Mendrinos S, Lance RA, 

Nyalwidhe JO, Beydoun HA, Clements MA, 

Drake RR, Semmes OJ (2009) 

Imaging mass spectrometry of a specific 

fragment of mitogen-activated protein kinase/ 

extracellular signal-regulated kinase kinase 

kinase 2 discriminates cancer from uninvolved 

prostate tissue. 

Clin Cancer Res. 15(17):5541-51 

Prostate cancer is a cancer type in which the 

differentiation of begning tissue and cancrous 

tissue may be difficult. In this study prostate 

cancer and normal tissue was differentiated by 

MALDI Imaging. The original findings in a small 

discovery set were verified by the analysis of 

a validation set with 54 tissue sections. One 

marker that correlated with the tumor was

identified as a fragment of mitogen-activated 

protein kinase/extracellular signal regulated 

kinase kinase kinase 2 (MEKK2). 

Groseclose MR, Massion PP, Chaurand P, 


High-throughput proteomic analysis of formalinfixed 

paraffin-embedded tissue microarrays 

using MALDI Imaging mass spectrometry. 

Proteomics. 8(18):3715-24 

The analysis of tissue microarrays is of great 

interest, since it allows the analysis of a large 

number of samples in a relatively short time. 

This article describes how MALDI Imaging 

can be used to analyze FFPE tissue by tryptic 

digestion and how tumor types can be classified 

based on their peptide patterns. The result 

of the classification of the individual pixels is 

visualized as image („Class imaging“). 

Lemaire R, Menguellet SA, Stauber J, 

Marchaudon V, Lucot JP, Collinet P, 

Farine MO, Vinatier D, Day R, Ducoroy P, 

Salzet M, Fournier I (2007) 

Specific MALDI Imaging and profiling for 

biomarker hunting and validation: fragment of 

the 11S proteasome activator complex, Reg 

alpha fragment, is a new potential ovary cancer 

biomarker. 


Early paper that shows the power of MALDI 

Imaging in clincial proteomic research for the 

analysis of ovary cancer. A specific 84 amino 

acid fragment from the protein Reg-alpha was 

identified as tumor marker and validated with 

other techniques, including immunohistochemsitry. 

Figure 2: Class Imaging: Visual representation of the statistical 

classification of four biopsies compared to the assignment based on 

histology. From: Groseclose et. al (2008),Proteomics 8(18) 3715 

© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim 

Drug imaging / Xenobiotics / 

Pharmaceutical research 

MALDI Imaging can be used to directly 

measure drugs, metabolites and xenobiotics in 

tissue 

Seppala U, Francese S, Turillazzi S, Moneti G, 

Clench M, Barber D (2011) 

In situ imaging of honeybee (Apis mellifera) 

venom components from aqueous and aluminum 

hydroxide-adsorbed venom immunotherapy 

preparations. 

J Allergy Clin Immunol. 129(5):1314-1320.e3 

An article that utilised MALDI Imaging to 

assess purified aqueous and aluminum hydroxide 

(Al[OH]3)adsorbed purified honeybee 

(Apis mellifera) venom (HBV) immunotherapy 

preparations in situ, which can reduce the incidence 

of side effects associated with venom 

immunotherapy. The study assessed the 

distribution of HBV components and showed 

that the majority of low-molecular-weight HBV 

components are rapidly removed from the site 

of venom immunotherapy administration. Furthermore, 

Al(OH)3-adsorbed HBV preparation 

demonstrated a depot effect, prolonging the 

availability of bee venom allergens at the site of 

administration. 

Goodwin RJ, Mackay CL, Nilsson A, Harrison 

D, Farde L, Andren PE, Iverson SL (2011) 

Qualitative and quantitative MALDI Imaging 

of the positron emission tomography ligands 

raclopride (a D2 dopamine antagonist) and 

SCH 23390 (a D1 dopamine antagonist) in rat 

brain tissue sections using a solvent-free dry 

matrix application method. 

Anal Chem. 83(24):9694-701 

An article that describes a dry matrix preparation 

for the detection of two dopamine receptor 

antagonists. Rats were dosed at 5 mg/kg and 

the distribution of the drug in the brain could be 

detected. 

Enthaler B, Pruns JK, Wessel S, Rapp C, 

Fischer M, Wittern KP (2011) 

Improved sample preparation for MALDI-MSI 

of endogenous compounds in skin tissue sections 

and mapping of exogenous active compounds 

subsequent to ex-vivo skin penetration. 

Anal Bioanal Chem. 402(3):1159-67 

In this article, MALDI Imaging was utilised for 

the determination of the pathway and depth of 

penetration of an exogenous model compound, 

Nile Red, after ex-vivo penetration into 

porcine skin. Additionally, the localisation of

endogenous compounds, including cholesterol 

sulphate, was measured directly on tissue at a 

resolution of approximately 30 m. 

Miki A, Katagi M, Kamata T, Zaitsu K, 

Tatsuno M, Nakanishi T, Tsuchihashi H, 

Takubo T, Suzuki K (2011) 

MALDI-TOF and MALDI-FTICR Imaging mass 

spectrometry of methamphetamine incorporated 

into hair. 

J Mass Spectrom. 46(4):411-6 

This paper describes the potential use of 

MALDI Imaging in forensics. Cocaine was 

measured directly in human hair and the spatial 

profile of cocaine in the hair could be translated 

to a temporary consumption profile. 

Marshall P, Toteu-Djomte V, Bareille P, Perry H, 

Brown G, Baumert M, Biggadike K (2010) 

Correlation of skin blanching and percutaneous 

absorption for glucocorticoid receptor agonists 

by matrix-assisted laser desorption ionization 

mass spectrometry imaging and liquid extraction 

surface analysis with nanoelectrospray 

ionization mass spectrometry. 

Anal Chem. 82(18):7787-94 

In this study, the percutaneous absorption of 

three novel nonsteroid glucocorticoid receptor 

agonists has been investigated. 

Nilsson A, Fehniger TE, Gustavsson L, 

Andersson M, Kenne K, Marko-Varga G, 

Andren PE (2010) 

Fine mapping the spatial distribution and 

concentration of unlabeled drugs within tissue 

micro-compartments using imaging mass 

spectrometry. 

PLoS One. 5(7):e11411 

This article describes the use of MALDI 

Imaging for monitoring in vivo transport of 

unlabelled drugs within specific organ tissue 

compartments. A reference compound (tiotropium) 

would be tracked and quantified within 

the lungs of dosed rats, using systematic pointby-point 

MS and MS/MS sampling across the 

tissue. 

Francese S, Lambardi D, Mastrobuoni G, 

la Marca G, Moneti G, Turillazzi S (2009) 

Detection of honeybee venom in envenomed 

tissues by direct MALDI MSI. 

J Am Soc Mass Spectrom. 20(1):112-23 

In this study, MALDI Imaging has been used 

to map the diffusion and distribution of venom 

allergen and toxins in skin. 

Acquadro E, Cabella C, Ghiani S, Miragoli L, 

Bucci EM, Corpillo D (2009) 

Matrix-assisted laser desorption ionization 

imaging mass spectrometry detection of a 

magnetic resonance imaging contrast agent in 

mouse liver. 

Anal Chem. 81(7):2779-84 


Imaging to detect a magnetic resonance 

imaging (MRI) contrast agent in both frozen and 

paraformaldehyde-fixed mouse livers after its in 

vivo administration. 

Dekker LJ, van Kampen JJ, Reedijk ML, 

Burgers PC, Gruters RA, Osterhaus AD, 

Luider TM (2009) 

A mass spectrometry based imaging method 

developed for the intracellular detection of HIV 

protease inhibitors. 

Rapid Commun Mass Spectrom. 23(8):1183-8 

HIV protease inhibitors have been measured in 

cultured cells using MALDI-TOF and MALDI- 

FTMS. 

Cornett DS, Frappier SL, Caprioli RM (2008) 

MALDI-FTICR Imaging mass spectrometry of 

drugs and metabolites in tissue. 

Anal Chem. 80(14):5648-53 

In this work, the authors show how the high 

precision of FTMS mass measurements 

increase the confidence in the results of drug 

imaging experiments. 

Meistermann H, Norris JL, Aerni HR, 

Cornett DS, Friedlein A, Erskine AR, 

Augustin A, De Vera Mudry MC, Ruepp S, 

Suter L, Langen H, Caprioli RM, Ducret A 

(2006) 

Biomarker discovery by imaging mass spectrometry: 

transthyretin is a biomarker for gentamicin-induced 

nephrotoxicity in rat. 

Mol Cell Proteomics. 5(10):1876-86 

In this article, MALDI Imaging has been used 

to find a surrogate biomarker in kidney that indicates 

the nephrotoxic effect of an administered 

drug, gentamicin.

Model Organisms 

A selection of studies that have used MALDI 

Imaging for general biological or biomedical 

research on model organisms 

Qureshi A, Subathra M, Grey A, Schey K, 

Del Poeta M, Luberto C (2010) 

Role of sphingomyelin synthase in controlling 

the antimicrobial activity of neutrophils against 

Cryptococcus neoformans. 

PLoS One. 5(12):e15587 

This study performed MALDI Imaging on lungs 

infected with Cryptococcus neoformans to 

provide new insights into the key host cellular 

pathway(s) necessary to control infection 

caused from inhalation of Cryptococcus neoformans. 

MALDI Imaging revealed specific lipid 

(SM - sphingomyelin) species are associated 

with neutrophil infiltration at the site of the 

infection. 

Magalhaes BS, Melo JA, Leite JR, Silva LP, 

Prates MV, Vinecky F, Barbosa EA, Verly RM, 

Mehta A, Nicoli JR, Bemquerer MP, 

Andrade AC, Bloch C Jr (2008) 

Post-secretory events alter the peptide content 

of the skin secretion of Hypsiboas raniceps. 

Biochem Biophys Res Commun. 377(4):1057- 

61 

In this article, MALDI Imaging demonstrated 

that mature raniseptin peptides (a novel family 

of antimicrobial peptides characterised from 

skin secretion of the anuran Hypsibas raniceps) 

are in fact secreted as intact molecules within a 

defined glandular domain of the dorsal skin, 

Figure 3: Statistical Analysis: Spatial segmentation map based on 

hierarchical clustering of a rat whole body section and principal component 

analysis of selected organs. From: Speir. et al. Proceedings of 

the 60th ASMS Conference on MassSpectrometry and Allied Topics, 

May 20-24,2012 

challenging the physiological role of the observed 

raniseptin fragments, identified only as part 

of the crude secretion. 

Brand GD, Krause FC, Silva LP, Leite JR, 

Melo JA, Prates MV, Pesquero JB, Santos EL, 

Nakaie CR, Costa-Neto CM, Bloch C Jr (2006) 

Bradykinin-related peptides from Phyllomedusa 

hypochondrialis. 

Peptides. 27(9):2137-46 link 

Frog mucus contains biologically active peptides 

that are e.g. protective of fungal infections 

or predators. MALDI Imaging has been used to 

map such peptides directly in frog skin. 

Neuroscience 

Selected MALDI Imaging papers in neuroscience 

Hanrieder J, Karlsson A, Falth M, Eriksson- 

Mammo S, Bergquist J, Andersson M (2011) 

L-DOPA-induced dyskinesia is associated with 

regional increase of striatal dynorphin peptides 

as elucidated by imaging mass spectrometry. 

Mol Cell Proteomics. 10(10):M111.009308 

MALDI Imaging was used for characterization, 

localization, and relative quantification of striatal 

neuropeptides in a rat model of l-DOPA-induced 

dyskinesia (LID) in Parkinson‘s disease. 

Yuki D, Sugiura Y, Zaima N, Akatsu H, 

Hashizume Y, Yamamoto T, Fujiwara M, 

Sugiyama K, Setou M (2011) 

Hydroxylated and non-hydroxylated sulfatide 

are distinctly distributed in the human cerebral 

cortex. 

Neuroscience. 193:44-53 

This article studies the distribution of sulfatide 

lipids in human cerebral cortex. It was found 

that hydroxylated and non-hydroxylated sulfatides 

are differently distributed in white and 

grey matter. An alzheimer brain has also been 

investigated and showed a similar distribution 

of sulfatides compared to normal tissue. 

Yang H, Sugiura Y, Ikegami K, Konishi Y, 

Setou M (2011) 

Axonal gradient of arachidonic acid-containing 

phosphatidylcholine and its dependence on 

actin dynamics. 

J Biol Chem. 287(8):5290-300 

This article used MALDI Imaging to characterise 

the distribution of phosphatidylcholine (PC) 

species in cultured neurons of superior cervical 

ganglia.

Sugiura Y, Taguchi R, Setou M (2011) 

Visualization of spatiotemporal energy 

dynamics of hippocampal neurons by mass 

spectrometry during a kainate-induced seizure. 

PLoS One. 6(3):e17952 

In this paper, MALDI Imaging combined with 

capillary electrophoresis (CE) mass spectrometry 

were utilised to visualise energy metabolism 

by imaging energy-related metabolites 

in mouse brains to reveal the spatiotemporal 

energy metabolism of the mouse hippocampus. 

Sugiura Y, Shimma S, Konishi Y, Yamada MK, 

Setou M (2008) 

Imaging mass spectrometry technology and 

application on ganglioside study; visualization 

of age-dependent accumulation of C20-ganglioside 

molecular species in the mouse hippocampus. 

PLoS One. 3(9):e3232 

The distrubtion of ganglioside molecular 

species in the brain was revealed by MALDI 

Imaging, enabling discrimination of species 

raised from both oligosaccharide and ceramide 

structure by the difference in mass-to-charge 

ratio and structural elucidation by MSMS. 

Plants 

MALDI Imaging in the investigation of plants 

Poth AG, Mylne JS, Grassl J, Lyons RE, Millar 

AH, Colgrave ML, Craik DJ (2012) 

Cyclotides associate with leaf vasculature and 

are the products of a novel precursor in Petunia 

(Solanaceae). 

J Biol Chem. doi:10.1074/jbc.M112.370841 

Cyclotides are defense related cylic peptides. In 

this study the spatial distribution of cyclotides 

was assessed by MALDI Imaging. The major 

cyclotide component showed a non-uniform 

distribution in petunia leaves. 

Holscher D, Shroff R, Knop K, Gottschaldt M, 

Crecelius A, Schneider B, Heckel DG, 

Schubert US, Svatos A (2009) 

Matrix-free UV-laser desorption/ionization (LDI) 

mass spectrometric imaging at the single-cell 

level: distribution of secondary metabolites of 

Arabidopsis thaliana and Hypericum species. 

Plant J. 60(5):907-18 

A paper that describes the direct analysis 

of secondary metabolites in Hypericum and 

Arabodopsis. The distribution of Naphtodianthrones 

were measured in Hypericum leavesm 

placenta, stamens and styli. The highest achieved 

spatial resolution was 10 micrometers. 

MALDI Imaging methodology 

MALDI Imaging is a field that is under research 

itself. This section shows interesting ongoing 

development in this field 

Thomas A, Charbonneau JL, Fournaise E, 

Chaurand P (2012) 

Sublimation of new matrix candidates for high 

spatial resolution imaging mass spectrometry 

of lipids: enhanced information in both positive 

and negative polarities after 1,5-diaminonapthalene 

deposition. 

Anal Chem. 84(4):2048-54 

As matrix sublimation has demonstrated to 

be a powerful approach for high-resolution 

MALDI Imaging of lipids, this study aimed to 

evaluate current and novel matrix candidates 

for high spatial resolution MALDI Imaging mass 

spectrometry of lipids from tissue section after 

deposition by sublimation. 

Angel PM, Spraggins JM, Baldwin HS, 

Caprioli R (2012) 

Enhanced sensitivity for high spatial resolution 

lipid analysis by negative ion mode matrix assisted 

laser desorption ionization imaging mass 

spectrometry. 

Anal Chem. 84(3):1557-64 

A paper that shows high spatial resolution of 10 

micrometer with an Ultraflextreme on tissue. 

Yang J, Caprioli RM (2011) 

Matrix sublimation/recrystallization for imaging 

proteins by mass spectrometry at high spatial 

resolution. 

Anal Chem. 83(14):5728-34 

A paper that investigates improved sample 

preparation methods based on matrix sublimation. 

It shows the technical possibilities of 

resolution obtainable with Brukers advanced 

instrumentation. 

Khatib-Shahidi S, Andersson M, Herman JL, 

Gillespie TA, Caprioli RM (2006) 

Direct molecular analysis of whole-body animal 

tissue sections by imaging MALDI mass spectrometry. 

Anal Chem. 78(18):6448-56 

A paper that shows how different substance 

classes can be measured in whole body sections 

of rats.

Method - Computational 

Computational methods are an important part 

of MALDI Imaging analysis. This section lists 

interesting papers in this field 

Trede D, Schiffler S, Becker M, Wirtz S, 

Steinhorst K, Strehlow J, Aichler M, Kobarg 

JH, Oetjen J, Dyatlov A, Heldmann S, Walch A, 

Thiele H, Maass P, Alexandrov T (2012) 

Exploring Three-Dimensional Matrix-Assisted 

Laser Desorption/Ionization Imaging Mass 

Spectrometry Data: Three-Dimensional Spatial 

Segmentation of Mouse Kidney. 

Anal Chem. doi:10.1021/ac300673y 

This paper describes an efficient computational 

pipeline which allows analysis and interpretation 

of three dimensional MALDI Imaging 

datasets. 

Andersson M, Groseclose MR, Deutch AY, 


Imaging mass spectrometry of proteins and 

peptides: 3D volume reconstruction. 

Nat Methods. 5(1):101-8 

The article presents an optimised step-by-step 

procedure describing how to make 3D volume 

reconstructions of MALDI Imaging data, as 

applied to the rat brain. 

Sinha TK, Khatib-Shahidi S, Yankeelov TE, 

Mapara K, Ehtesham M, Cornett DS, Dawant 

BM, Caprioli RM, Gore JC (2008) 

Integrating spatially resolved three-dimensional 

MALDI IMS with in vivo magnetic resonance 

imaging. 

Nat Methods. 5(1):57-9 

The article describes a method for integration 

of 3D volume reconstructions of proteomic 

MALDI Imaging datasets of whole mouse 

heads with in vivo high-resolution images from 

other imaging modalities (magnetic resonance 

imaging). 

Hanselmann M, Kirchner M, Renard BY, 

Amstalden ER, Glunde K, Heeren RM, 

Hamprecht FA (2008) 

Concise representation of mass spectrometry 

images by probabilistic latent semantic analysis. 

Anal Chem. 80(24):9649-58 

This paper describes a very interesting statistical 

method, named probabibilistic latent 

sematics analysis (pLSA), for the concise representation 

of complex imaging mass spectrometry 

data. Although the data was not recorded 

on Bruker instrumentation,In the meantime, 

pLSA has been implemented in Brukers MALDI 

imaging software package. 

Figure 4: Drug Imaging / Histopharmacology: Sagittal rabbit brain section showing distribution of a cysteine 

conjugate metabolite. (A) Monochromatic ion image showing brain distribution of cysteine conjugate metabolite 

(170 µm spatial resolution). (B) Same ion image shown in rainbow intensity format. (C) Hematoxylin and eosin 

histology slide showing regions of the rabbit brain. From: Castellino et al. (2011) Bioanalysis 3(21),2427-41 with 

kind permission of the authors.

Development 

MALDI imaging papers in developmental 

biology 

Lagarrigue M, Becker M, Lavigne R, 

Deininger SO, Walch A, Aubry F, Suckau D, 

Pineau C (2011) 

Revisiting rat spermatogenesis with MALDI 

Imaging at 20-microm resolution. 

Mol Cell Proteomics. 10(3):M110.005991 

This article investigates the protein expression 

in rat testis. It shows state-of-the-art protein 

images at a spatial resolution of 20 micrometer. 

The observed protein profiles were linked to 

the stages of spermatogenesis as determined 

by histology. The article therefore shows the 

importance of the correlation of molecular 

MALDI images and histology. Some of the 

observed proteins were identified and their 

expression further investigated by immunohistochemistry. 

Goto-Inoue N, Hayasaka T, Zaima N, Setou M 

(2009) 

The specific localization of seminolipid molecular 

species on mouse testis during testicular 

maturation revealed by imaging mass spectrometry. 

Glycobiology. 19(9):950-7 

MALDI Imaging was utilised to show cell-specific 

localisation of each molecular species of 

seminolipid (a unique sulphated glyceroglycolipid, 

which constitutes more than 90% of the 

glycolipid in mammalian testis) during testicular 

maturation. 

Burnum KE, Cornett DS, Puolitaival SM, 

Milne SB, Myers DS, Tranguch S, Brown HA, 

Dey SK, Caprioli RM (2009) 

Spatial and temporal alterations of phospholipids 

determined by mass spectrometry during 

mouse embryo implantation. 

J Lipid Res. 50(11):2290-8 

MALDI Imaging was used to characterise 

the spatial patial and temporal distribution of 

phospholipid species associated with mouse 

embryo implantation. LC-ESI-MSMS validated 

the phospholipid distribution patterns observed 

using MALDI Imaging. 

Burnum KE, Tranguch S, Mi D, Daikoku T, Dey 

SK, Caprioli RM (2008) 

Imaging mass spectrometry reveals unique 

protein profiles during embryo implantation. 

Endocrinology. 149(7):3274-8 

Chaurand P, Rahman MA, Hunt T, Mobley JA, 

Gu G, Latham JC, Caprioli RM, Kasper S (2008) 

Monitoring mouse prostate development by 

profiling and imaging mass spectrometry. 

Mol Cell Proteomics. 7(2):411-23 


Imaging to evaluate protein expression profiles 

in the normal mouse prostate during development, 

at sexual maturation and in adult stages. 

Microbiology 

Interesting papers of the use of MALDI Imaging 

to study microbial interactions 

Liu WT, Kersten RD, Yang YL, Moore BS, 

Dorrestein PC (2011) 

Imaging mass spectrometry and genome 

mining via short sequence tagging identified 

the anti-infective agent arylomycin in Streptomyces 

roseosporus. 

J Am Chem Soc. 133(45):18010-3 

A paper that utilised MALDI Imaging along with 

a peptidogenomic approach (expanded to short 

sequence tagging SST) for discovery of antiinfective 

agent arylomycin and its biosynthetic 

gene cluster in an industrial daptomycin producing 

strain Streptomyces roseosporus. 

Liu WT, Yang YL, Xu Y, Lamsa A, Haste NM, 

Yang JY, Ng J, Gonzalez D, Ellermeier CD, 

Straight PD, Pevzner PA, Pogliano J, Nizet V, 

Pogliano K, Dorrestein PC (2010) 

Imaging mass spectrometry of intraspecies 

metabolic exchange revealed the cannibalistic 

factors of Bacillus subtilis. 

Proc Natl Acad Sci U S A. 107(37):16286-90 

This paper studies molecular interactions in 

bacterial cannibalism. 

Yang YL, Xu Y, Straight P, Dorrestein PC (2009) 

Translating metabolic exchange with imaging 

mass spectrometry. 

Nat Chem Biol. 5(12):885-7 

In this article the metabolic interaction of 

different bacterial strains on an agar plate are 

monitored by MALDI Imaging. This allowed a 

direct molecular characterization of a chemical 

fight between the bacterial strains and provided 

evidence that Bacillus subtilis silenced the 

defensive arsenal of Streptomyces coelicolor.

— 200 µm 

m/z726.5 PE 36:2 

m/z774.6 PE-p 40:6 

m/z885.6 PI 38:4 

Figure 5: Lipids in mouse cerebellum measured at 10 µm spatial resolution. 

Reprinted from: Thomas A, Charbonneau JL, Fournaise E, Chaurand P. Sublimation of new matrix 

candidates for high spatial resolution imaging mass spectrometry of lipids: enhanced information in both 

positive and negative polarities after 1,5-diaminonapthalene deposition. Anal Chem. 2012;84(4):2048-54 

Copyright (2012) American Chemical Society 

Sample Preparation 

m/z664.6 CerP 18:0 

m/z747.5 PG 34:1 

m/z786.5 PS 36:2 

m/z890.6 ST 24:0 

Data Acquisition 

Figure 6: Bruker offers the complete MALDI Imaging solution. 

m/z700.5 PE-p 34:1 

m/z762.6 PE 38:6 

m/z834.6 PS 40:6 

m/z700.5 + m/z 786.5 

m/z715.6 PA-o 38:1 

m/z766.6 PE 38:4 

m/z878.6 ST 22:0 (OH) 

m/z834.6 + m/z 890.6 

Data Evaluation 

Statistical Analysis

Bruker Author 

Bruker does actively contribute to research in 

the MALDI Imaging field, having introduced 

key innovations in both hard- and software. 

This section lists papers with key conttributions 

form Bruker authors 

Deininger SO, Cornett DS, Paape R, Becker M, 

Pineau C, Rauser S, Walch A, Wolski E (2011) 

Normalization in MALDI-TOF Imaging datasets 

of proteins: practical considerations. 

Anal Bioanal Chem. 401(1):167-81 

Normalization is an important step in the data 

preparation of MALDI Imaging data, mostly to 

account random real-world influences in the 

data. Commonly used normalization techniques 

such as the total ion count, can introduce 

artifacts in rare cases. This paper discusses the 

applicability of different normalization approaches 

and introduces robust methods, such 

as normalization to median intensity. These 

normalization options are included in Brukers 

MALDI Imaging software. 

Alexandrov T, Becker M, Deininger SO, 

Ernst G, Wehder L, Grasmair M, 

von Eggeling F, Thiele H, Maass P (2010) 

Spatial segmentation of imaging mass spectrometry 

data with edge-preserving image 

denoising and clustering. 


This paper describes how MALDI Imaging data 

can be improved by spatially aware de-noising. 

Compared to mere smoothing of the images, 

edge-preserving de-noising can improve 

MALDI images, while keeping highly resolved 

fine structures in the images. 

Agar NY, Kowalski JM, Kowalski PJ, Wong JH, 

Agar JN (2010) 

Tissue preparation for the in situ MALDI MS 

Imaging of proteins, lipids, and small molecules 

at cellular resolution. 

Methods Mol Biol. 656:415-31 

For research use only. Not for use in diagnostic procedures. 

Bruker Daltonik GmbH 

Bremen · Germany 

Phone +49 (0)421-2205-0 

Fax +49 (0)421-2205-103 

sales@bdal.de 

www.bruker.com/maldiimaging 

Bruker Daltonics Inc. 

This chapter presents three methods that 

minimise the delocalisation of analytes during 

matrix deposition, including a method where 

image resolution is not limited by the laser 

focal diameter. 

Deininger SO, Ebert MP, Futterer A, 

Gerhard M, Rocken C (2008) 

MALDI Imaging combined with hierarchical 

clustering as a new tool for the interpretation 

of complex human cancers. 


This paper describes how hierarchical clustering 

can be used to do a highly interactive and 

consice detailed annotation of MALDI Imaging 

datasets. The user interface for the clustering 

and the software to perform the analysis is 

part of Brukers MALDI Imaging software. 

Taban IM, Altelaar AF, van der Burgt YE, 

McDonnell LA, Heeren RM, Fuchser J, 

Baykut G (2007) 

Imaging of peptides in the rat brain using 

MALDI-FTICR mass spectrometry. 

J Am Soc Mass Spectrom. 18(1):145-51 

The first paper that shows MALDI Imaging 

with very high mass spectrometric resolution 

on FTMS instrumentation. 

Holle A, Haase A, Kayser M, Hohndorf J 

(2006) 

Optimizing UV laser focus profiles for improved 

MALDI performance. 

J Mass Spectrom. 41(6):705-16 

The laser is an important part of the MALDI 

process; even being contained in the term 

„MALDI“. Brukers proprietary smartbeam 

laser is widely recognized for its superior 

performance, softness of ionization and spatial 

resolution. This paper describes the principle 

and performance advantages of the smartbeam 

laser. 

Billerica, MA · USA Fremont, CA · USA 

Phone +1 (978) 663-3660 Phone +1 (510) 683-4300 

Fax +1 (978) 667-5993 Fax +1 (510) 490-6586 

ms-sales@bdal.com ms-sales@bdal.com 

Bruker Daltonics is continually improving its products and reserves the right 

to change specifications without notice. © BDAL 06-2012, #701576

Selected MALDI Imaging Publications - Bruker Daltonics

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