HTX Applications

  • Forensics
  • Environmental
  • Proteomics
  • Drug development

HTX Applications

Forensic Applications

Application of MALDI MS Imaging to Forensic Science

It is essential that forensic researchers utilize highly effective analytical techniques that produce results with the highest level of confidence so that evidence can be upheld in court.  Thanks to the large dynamic range of MALDI Mass Spectrometers, researchers are able to analyze a wide range of compounds in unknown samples.  Information on both endogenous and exogenous compounds can be obtained in a single experiment without extensive sample separation and preparation.


MALDI MS, a Natural Fit for Forensic Labs

Forensic labs already rely heavily on Mass Spectrometry (MS) to analyze trace compounds on solid samples (e.g.:  GC/MS analysis of explosives) or body fluids (e.g.: LC/MS analysis of urine and blood for drug of abuse detection), and such test results, are fully acceptable in court.  Forensic labs also rely routinely on DNA test kits to match suspects with crime scene evidence.  MALDI Mass Spectrometry, with its ability to directly analyze intact bio-molecules that contain genetic information (e.g.: proteins), as well as a wide range of other chemicals, is therefore, a natural fit for forensic labs.


The Power of MALDI MS Imaging

Scientists began applying MALDI MS Imaging to the analysis of forensic evidence and are now able to obtain new types of valuable information.  In a recent study, Dr Simona Francese at Sheffield-Hallam University (UK), working within the team of Professor Malcolm Clench, has used MALDI MS Imaging to capture fingerprint ridge patterns simultaneous to the detection of embedded compounds.  The study shows how the analytical power of mass spectrometry can be used in sexual assault cases to detect both endogenous species (e.g.: oleic acid), as well as exogenous species (e.g.: condoms lubricants) in fingerprints, and consequently, can link a suspect to a crime scene and other criminal misconduct.

HTX TECH NOTE #32   A Rapid Methodology for the Analysis of Latent Fingermarks by MALDI MS Imaging

HTX Tech Note #32 Sheffield-Hallam team develop a Rapid Methodology for the Analysis of Latent Fingermarks by MALDI MS Imaging
  Examples of endogenous species imaging (Images provided by Drs. Rob Bradshaw and Simona Francese, Sheffield-Hallam University).

Examples of endogenous species imaging (Images provided by Drs. Rob Bradshaw and Simona Francese, Sheffield-Hallam University).

Environmental Applications

APPLICATION OF MALDI MS IMAGING TO ENVIRONMENTAL SCIENCE

MALDI Mass Spectrometry (MS) is a great analytical platform for environmental science because it offers excellent sensitivity over a large dynamic range and rapid analysis time.  In one single experiment, scientists are able to obtain information on a wide range of compounds, known and unknown, without the need for prior labeling.  Both man made compounds and natural environmental compounds can be detected and analyzed.  Practical application include monitoring know substances in wildlife and studying the localization of compounds throughout the animal body.


US EPA biologist uses the TM-Sprayer to produce Mass Spectrometry imaging of proteins in Zebra Fish

TM-Sprayer was used to deposit a matrix solution with 5 mg/ml SPA, 250 ul/min flow rate, 1200 velocity, 1 min dry time between passes, with 8 passes, 90 degree turn between each pass and 1 mm offset.

Proteins present in ovaries and eggs are imaged by Mass spectrometry imaging (AB Sciex MALDI TOF 4800)

Proteins present in ovaries and eggs are imaged by Mass spectrometry imaging (AB Sciex MALDI TOF 4800)


COMPOSITION AND SPATIAL ANALYSIS IN ONE EXPERIMENT

Mass Spectrometry offers the ability to provide compositional information on a wide range of compounds such as animal proteins without the need to first generate specific antibodies.  This can help scientists provide rapid answers following an environmental crisis.

Mapping of Bio-Molecules in Animal Organs

In a paper published in 2009, Ruibing Chen et al showed how MALDI MS Imaging can be used to produce 3D mapping of bio-molecules in a Cancer borealis crab brain.  The identification and confirmation of key lipids was achieved by using the MS/MS fragmentation.

Three Dimensional Mapping of Neuropeptides and Lipids in Crustacean Brain by Mass Spectral Imaging

Proteomic Applications

APPLICATION OF MALDI MASS SPECTROMETRY TO PROTEOMICS DISCOVERY

MALDI Mass Spectrometry (MS) is a proven analytical platform for proteomics discovery that offers excellent sensitivity and rapid analysis time. In one single experiment, scientists are able to obtain information on a wide range of compounds, known and unknown, without the need for prior labeling. The MALDI source is especially well suited to proteomics because large molecules such as proteins, peptides and lipids can be difficult to analyze without damaging their structure. The soft ionization process of MALDI provides the ability to study intact bio-molecules and when combined to the power of tandem mass spectrometry offers the analytical resolution needed to explicit complex molecular structures.


HOW MALDI MS IMAGING CHANGES OUR UNDERSTANDING OF BIOLOGICAL COMPOSITION

By using Mass Spectrometry data to create molecular images scientists are able to visualize the spatial distribution of proteins and other bio-molecules.  Thanks to the wide dynamic range of Mass Spectrometers, each experiment actually produces a multitude of images and the ability to visualize multiple species at once.

In the following example, a sagital section of a rat brain has been analyzed by MALDI MS Imaging to produce high resolution maps for species with mass range <1,000 Daltons.  Each specie is characterized by its m/z (mass/ion charge) ratio and an imaging software is used to render relative concentration based on a luminosity gradient (Images A-F).  Using the same data, a user can also produce a multicolor image (Image G) where one specie is represented in blue and the other in red, while still using a gradient to render relative concentration.

Further analysis is done to identify species of interest and this information can then be used in future work to formally identify specific species.  In the images below (H-J and K-N) different lipids have been identified.  From these images, one can clearly see that M 385.4 [Cholesterol – H]+ is only present in certain areas of the brain, whereas M 429.3 [Vitamin E –H]+ is present throughout the tissue.  An optical microscope image of the tissue (O) is used to verify the correspondence of Mass Spectrometric data and with histological data.

Drug Development Applications

APPLICATION OF MALDI MASS SPECTROMETRY IMAGING (MSI) TO DRUG DEVELOPMENT

MALDI MS Imaging is a powerful technique that is especially useful for researchers in the pharmaceutical field. Researchers can visualize the effects of new therapies by determining the precise distribution of drugs and drug metabolites in a tissue section. Researchers can also extend their understanding of therapeutic pathways by comparing the proteomic make up of healthy and diseased organisms.


IMAGING OF DRUG METABOLISM WITHOUT RADIOACTIVE ISOTOPE LABELING

The ability to detect molecules with mass ranges up to 500,000 Daltons makes mass spectrometry a favored detection technique for drug research.  In addition, the precise characterization of bio-markers, without the need for radio isotope labeling or tagging, simplifies processes and enables de novo identification.

“Top-Down” analysis:  Direct measure of the mass (m/z) of intact, small or large, bio-molecules and possible analysis of molecule fragments for additional structure information.

“Bottom-Up” analysis:  Direct measure of the mass (m/z) of fragmented bio-molecules and analysis of these fragment molecules by LC/MS or MALDI/MS to identify the original molecule.  A typical application is the digestion of proteins into peptides (e.g.: trypsin digestion) and the de novo sequencing of the original protein based on the identification of its sub-components.


MAPPING LARGE QUANTITIES OF DATA INTO EASY TO INTERPRET IMAGES

By relating the analytical MALDI MS data to the X-Y coordinates of the MALDI laser at the time of analysis, MALDI MS Imaging provides researchers the ability to determine the exact distribution pattern of drugs and drug metabolites throughout a tissue section or an entire study animal section.  The MS data is easily exported into an imaging software package, such as NOVARTIS’ BioMap (Free download at www.msi-imaging.com), for image generation and presentation.


UNDERSTANDING BIOLOGICAL PATHWAYS AND VISUALIZING THE EFFECT OF POTENTIAL TREATMENTS

Understanding the distribution of specific proteins in a tissue, as well as other molecules such as lipids and metabolites, can be of great help to understand complex diseases, such as Alzheimer’s.  The “Molecular Images” generated by MALDI MS can be used as molecular signature for pathological progression, and therefore, be also used to evaluate the benefits of new therapies.


APPLICATIONS OF MALDI MS IMAGING IN DRUG DEVELOPMENT

  • Make earlier go/no-go decisions based on drug distribution.
  • Optimize drug delivery mechanism (E.g.:  Application to dermatology by observing the penetration pattern of drugs in epidermal tissue.  Selection of optimum carriers).
  • Identify compounds faster by obtaining accurate mass information directly from tissue samples.
  • Determine precise compound distribution by acquiring high-spatial resolution images.

METHOD DEVELOPMENT RESOURCES

HTX Imaging provides full method development services to help labs optimize sample preparation prior to MS Imaging.  Optimized sample preparation helps increase analytes extraction and signal intensity for improved Mass Spec data.  The unique ability to control spray temperature is key in accelerating extraction while reducing delocalisation.

In the example below, scientists at Sheffield-Hallam University and GSK optimized flow rate and stage velocity for improved extraction of a new drug candidate.  Spray nozzle temperature of 80 Deg C allows very fast sample preparation and flow rates 5 to 10 times higher than other available matrix sprayers.

Application of MALDI MS Imaging to Forensic Science

It is essential that forensic researchers utilize highly effective analytical techniques that produce results with the highest level of confidence so that evidence can be upheld in court.  Thanks to the large dynamic range of MALDI Mass Spectrometers, researchers are able to analyze a wide range of compounds in unknown samples.  Information on both endogenous and exogenous compounds can be obtained in a single experiment without extensive sample separation and preparation.


MALDI MS, a Natural Fit for Forensic Labs

Forensic labs already rely heavily on Mass Spectrometry (MS) to analyze trace compounds on solid samples (e.g.:  GC/MS analysis of explosives) or body fluids (e.g.: LC/MS analysis of urine and blood for drug of abuse detection), and such test results, are fully acceptable in court.  Forensic labs also rely routinely on DNA test kits to match suspects with crime scene evidence.  MALDI Mass Spectrometry, with its ability to directly analyze intact bio-molecules that contain genetic information (e.g.: proteins), as well as a wide range of other chemicals, is therefore, a natural fit for forensic labs.


The Power of MALDI MS Imaging

Scientists began applying MALDI MS Imaging to the analysis of forensic evidence and are now able to obtain new types of valuable information.  In a recent study, Dr Simona Francese at Sheffield-Hallam University (UK), working within the team of Professor Malcolm Clench, has used MALDI MS Imaging to capture fingerprint ridge patterns simultaneous to the detection of embedded compounds.  The study shows how the analytical power of mass spectrometry can be used in sexual assault cases to detect both endogenous species (e.g.: oleic acid), as well as exogenous species (e.g.: condoms lubricants) in fingerprints, and consequently, can link a suspect to a crime scene and other criminal misconduct.

HTX TECH NOTE #32   A Rapid Methodology for the Analysis of Latent Fingermarks by MALDI MS Imaging

HTX Tech Note #32 Sheffield-Hallam team develop a Rapid Methodology for the Analysis of Latent Fingermarks by MALDI MS Imaging

 

  Examples of endogenous species imaging (Images provided by Drs. Rob Bradshaw and Simona Francese, Sheffield-Hallam University).

 

Examples of endogenous species imaging (Images provided by Drs. Rob Bradshaw and Simona Francese, Sheffield-Hallam University).

APPLICATION OF MALDI MS IMAGING TO ENVIRONMENTAL SCIENCE

MALDI Mass Spectrometry (MS) is a great analytical platform for environmental science because it offers excellent sensitivity over a large dynamic range and rapid analysis time.  In one single experiment, scientists are able to obtain information on a wide range of compounds, known and unknown, without the need for prior labeling.  Both man made compounds and natural environmental compounds can be detected and analyzed.  Practical application include monitoring know substances in wildlife and studying the localization of compounds throughout the animal body.


US EPA biologist uses the TM-Sprayer to produce Mass Spectrometry imaging of proteins in Zebra Fish

TM-Sprayer was used to deposit a matrix solution with 5 mg/ml SPA, 250 ul/min flow rate, 1200 velocity, 1 min dry time between passes, with 8 passes, 90 degree turn between each pass and 1 mm offset.

Proteins present in ovaries and eggs are imaged by Mass spectrometry imaging (AB Sciex MALDI TOF 4800)

Proteins present in ovaries and eggs are imaged by Mass spectrometry imaging (AB Sciex MALDI TOF 4800)


COMPOSITION AND SPATIAL ANALYSIS IN ONE EXPERIMENT

Mass Spectrometry offers the ability to provide compositional information on a wide range of compounds such as animal proteins without the need to first generate specific antibodies.  This can help scientists provide rapid answers following an environmental crisis.

Mapping of Bio-Molecules in Animal Organs

In a paper published in 2009, Ruibing Chen et al showed how MALDI MS Imaging can be used to produce 3D mapping of bio-molecules in a Cancer borealis crab brain.  The identification and confirmation of key lipids was achieved by using the MS/MS fragmentation.

Three Dimensional Mapping of Neuropeptides and Lipids in Crustacean Brain by Mass Spectral Imaging

APPLICATION OF MALDI MASS SPECTROMETRY TO PROTEOMICS DISCOVERY

MALDI Mass Spectrometry (MS) is a proven analytical platform for proteomics discovery that offers excellent sensitivity and rapid analysis time. In one single experiment, scientists are able to obtain information on a wide range of compounds, known and unknown, without the need for prior labeling. The MALDI source is especially well suited to proteomics because large molecules such as proteins, peptides and lipids can be difficult to analyze without damaging their structure. The soft ionization process of MALDI provides the ability to study intact bio-molecules and when combined to the power of tandem mass spectrometry offers the analytical resolution needed to explicit complex molecular structures.


HOW MALDI MS IMAGING CHANGES OUR UNDERSTANDING OF BIOLOGICAL COMPOSITION

By using Mass Spectrometry data to create molecular images scientists are able to visualize the spatial distribution of proteins and other bio-molecules.  Thanks to the wide dynamic range of Mass Spectrometers, each experiment actually produces a multitude of images and the ability to visualize multiple species at once.

In the following example, a sagital section of a rat brain has been analyzed by MALDI MS Imaging to produce high resolution maps for species with mass range <1,000 Daltons.  Each specie is characterized by its m/z (mass/ion charge) ratio and an imaging software is used to render relative concentration based on a luminosity gradient (Images A-F).  Using the same data, a user can also produce a multicolor image (Image G) where one specie is represented in blue and the other in red, while still using a gradient to render relative concentration.

Further analysis is done to identify species of interest and this information can then be used in future work to formally identify specific species.  In the images below (H-J and K-N) different lipids have been identified.  From these images, one can clearly see that M 385.4 [Cholesterol – H]+ is only present in certain areas of the brain, whereas M 429.3 [Vitamin E –H]+ is present throughout the tissue.  An optical microscope image of the tissue (O) is used to verify the correspondence of Mass Spectrometric data and with histological data.

APPLICATION OF MALDI MASS SPECTROMETRY IMAGING (MSI) TO DRUG DEVELOPMENT

MALDI MS Imaging is a powerful technique that is especially useful for researchers in the pharmaceutical field. Researchers can visualize the effects of new therapies by determining the precise distribution of drugs and drug metabolites in a tissue section. Researchers can also extend their understanding of therapeutic pathways by comparing the proteomic make up of healthy and diseased organisms.


IMAGING OF DRUG METABOLISM WITHOUT RADIOACTIVE ISOTOPE LABELING

The ability to detect molecules with mass ranges up to 500,000 Daltons makes mass spectrometry a favored detection technique for drug research.  In addition, the precise characterization of bio-markers, without the need for radio isotope labeling or tagging, simplifies processes and enables de novo identification.

“Top-Down” analysis:  Direct measure of the mass (m/z) of intact, small or large, bio-molecules and possible analysis of molecule fragments for additional structure information.

“Bottom-Up” analysis:  Direct measure of the mass (m/z) of fragmented bio-molecules and analysis of these fragment molecules by LC/MS or MALDI/MS to identify the original molecule.  A typical application is the digestion of proteins into peptides (e.g.: trypsin digestion) and the de novo sequencing of the original protein based on the identification of its sub-components.


MAPPING LARGE QUANTITIES OF DATA INTO EASY TO INTERPRET IMAGES

By relating the analytical MALDI MS data to the X-Y coordinates of the MALDI laser at the time of analysis, MALDI MS Imaging provides researchers the ability to determine the exact distribution pattern of drugs and drug metabolites throughout a tissue section or an entire study animal section.  The MS data is easily exported into an imaging software package, such as NOVARTIS’ BioMap (Free download at www.msi-imaging.com), for image generation and presentation.


UNDERSTANDING BIOLOGICAL PATHWAYS AND VISUALIZING THE EFFECT OF POTENTIAL TREATMENTS

Understanding the distribution of specific proteins in a tissue, as well as other molecules such as lipids and metabolites, can be of great help to understand complex diseases, such as Alzheimer’s.  The “Molecular Images” generated by MALDI MS can be used as molecular signature for pathological progression, and therefore, be also used to evaluate the benefits of new therapies.


APPLICATIONS OF MALDI MS IMAGING IN DRUG DEVELOPMENT

  • Make earlier go/no-go decisions based on drug distribution.
  • Optimize drug delivery mechanism (E.g.:  Application to dermatology by observing the penetration pattern of drugs in epidermal tissue.  Selection of optimum carriers).
  • Identify compounds faster by obtaining accurate mass information directly from tissue samples.
  • Determine precise compound distribution by acquiring high-spatial resolution images.

METHOD DEVELOPMENT RESOURCES

HTX Imaging provides full method development services to help labs optimize sample preparation prior to MS Imaging.  Optimized sample preparation helps increase analytes extraction and signal intensity for improved Mass Spec data.  The unique ability to control spray temperature is key in accelerating extraction while reducing delocalisation.

In the example below, scientists at Sheffield-Hallam University and GSK optimized flow rate and stage velocity for improved extraction of a new drug candidate.  Spray nozzle temperature of 80 Deg C allows very fast sample preparation and flow rates 5 to 10 times higher than other available matrix sprayers.