Imaging Mass Microscope
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High-resolution imaging offered by optical microscopes is required not only for pharmacokinetic analysis, but also for toxicity testing and toxicity mechanism analysis. Analysis of the retina and skin requires imaging with high spatial resolution.
High spatial resolution imaging of mouse skin
In this experiment, the fat in rat skin was visualized. High spatial resolution imaging clearly showed localization of ceramide in the horny layer and phosphatidylcholine (PC) in the sebaceous gland.
Visualization of Lipid in a Mouse Cerebellum
A wide variety of molecules relate to diseases states. Imaging mass spectrometry using the iMScope TRIO can detect a wide range of molecules within a defined mass range. Therefore, distribution information for several target molecules with different molecular weights can be determined simultaneously during a single measurement. Visualization Profiling
By displaying the signal intensity at molecular weight of each target molecule, the 2D location of the target molecules is visualized. All target molecules can be detected simultaneously by a single measurement. Furthermore, samples are measured at a fast 6 pixels per second*, dramatically shortening your experiment time. In this example, the distribution of phosphatidylcholines (PCs) in the mouse cerebellum was successfully visualized within 3 hours (2.5 mm square). Limitations in immunostaining of lipids have previously made visual mapping of these compounds difficult. However this can easily be achieved using the iMScope TRIO, making this technology very powerful, particularly in areas such as brain function analysis and any biological process where lipids are known to play an important role.
High Spatial Resolution Imaging of Mouse Retina
High-resolution imaging offered by optical microscopes is required not only for pharmacokinetic analysis, but also for toxicity testing and toxicity mechanism analysis. Also, evaluating the phototoxic property of drug candidates requires detailed analysis of the retina and neighboring organs.
10 μm Imaging
In this experiment, two types of phosphatidylcholine (PC) distributions in the mouse retina were visualized. The PC (18:0/22:6) located next to the retinal pigment epithelium, which is about 10 μm thick, and the PC (16:0/18:1) located toward the outside the epithelium, can be detected and visualized distinctively. The images show that the two types of PC are distributed in thin layered structures.
Overlaying MS images and optical microscope morphological images reveals the difference between the amounts of specific molecules in each minute organ and can relate molecular distribution to the biological functions of an organelle and morphological changes.
The above figures show the resultant images generated by overlaying MS images at different m/z (blue and yellow) and optical microscope morphological images. These figures clearly indicate that analysis of the localization of specific molecules combined with morphological images is a useful approach for investigating drug efficiency and toxicity in drug discovery.
Seminolipids in mouse testis
Furthermore, samples are measured at a fast 6 pixels per second*, dramatically shortening your experiment time. In this example, the distribution of seminolipids in the mouse testis was successfully visualized within 3 hours (2.5 mm square).
*Ionization time: 50 ms,
mass range: m/z 500–1000, MS mode
The images show characteristic distributions of seminolipids (m/z 795.5 and 809.5) and phosphatidylinositols (m/z 885.5) in mouse testis. In this example, 9-aminoacridine was vapor-deposited as a matrix on the sample, and mass spectra were measured in negative ion mode. Negative ion spectra are simpler than positive ion, making it easier to identify target molecules. Negative ion analysis is frequently used for low molecular weight metabolite analysis.
In pharmacokinetic analysis, the iMScope TRIO enables the distribution of unchanged and metabolized drugs to be simultaneously mapped in a single measurement, without any labeling. The laser diameter used during mass spectrometry imaging with the iMScope TRIO is continuously variable from 5 to 200 μm, offering low to high spatial resolution. This helps ensure that analyses are performed as efficiently as possible.
Pharmacokinetic analysis of mouse with olaparib administered
Using a mouse administered with olaparib, the pharmacokinetic status of the tumor was visualized. In this way, imaging mass spectrometry is being utilized even in the clinical trial phase of drug discovery, rather than only during basic research.
Pharmacokinetic analysis of mouse with paclitaxel/micellar paclitaxel administered
This shows that micellation of the known anticancer agent paclitaxel improves its retention within the tumor. Consequently, imaging mass spectrometry is being utilized for DDS research as well.
Samples provided by the Division of Developmental Therapeutics,
Research Center for Innovative Oncology, National Cancer Center Hospital East
Visualize the molecular distribution (multiplexed imaging)
MS and MS/MS Imaging
Even if a molecule cannot be identified in the MS spectra, due to multiple molecules coexisting at the same mass (peak), these can be distinguished and visualized (rendered to images) by MS/MS analysis.
In industrial fields, various surface inspection technologies are widely used to ensure stable production of high quality products. Imaging mass spectrometry is capable of analyzing various contaminants that cannot be detected by conventional surface inspection methods, and it is possible to gain new knowledge for higher quality production.
An example using IC chip is shown on the left. A wide variety of optical images and mass spectrometry imaging technologies are used to support cutting-edge research.