Correlative mass spectrometry imaging, applying time-of-flight secondary ion mass spectrometry and atmospheric pressure matrix-assisted laser desorption/ionization to a single tissue section.

RATIONALE: Mass spectrometry imaging (MSI) is a powerful tool for mapping the surface of a sample. Time-of-flight secondary ion mass spectrometry (TOF-SIMS) and atmospheric pressure matrix-assisted laser desorption/ionization (AP-MALDI) offer complementary capabilities. Here, we present a workflow to apply both techniques to a single tissue section and combine the resulting data for the example of human colon cancer tissue. METHODS: Following cryo-sectioning, images were acquired using the high spatial resolution (1 µm pixel size) provided by TOF-SIMS. The same section was then coated with a para-nitroaniline matrix and images were acquired using AP-MALDI coupled to an Orbitrap mass spectrometer, offering high mass resolution, high mass accuracy and tandem mass spectrometry (MS/MS) capabilities. Datasets provided by both mass spectrometers were converted into the open and vendor-independent imzML file format and processed with the open-source software MSiReader. RESULTS: The TOF-SIMS and AP-MALDI mass spectra show strong signals of fatty acids, cholesterol, phosphatidylcholine and sphingomyelin. We showed a high correlation between the fatty acid ions detected with TOF-SIMS in negative ion mode and the phosphatidylcholine ions detected with AP-MALDI in positive ion mode using a similar setting for visualization. Histological staining on the same section allowed the identification of the anatomical structures and their correlation with the ion images. CONCLUSIONS: This multimodal approach using two MSI platforms shows an excellent complementarity for the localization and identification of lipids. The spatial resolution of both systems is at or close to cellular dimensions, and thus spatial correlation can only be obtained if the same tissue section is analyzed sequentially. Data processing based on imzML allows a real correlation of the imaging datasets provided by these two technologies and opens the way for a more complete molecular view of the anatomical structures of biological tissues.

[A prospective comparison of video colonoscopy and CT colonography in asymptomatic patients screened for colorectal cancer]. / CT-Kolonographie versus Videokoloskopie zum Nachweis kolorektaler Läsionen in der Vorsorgepopulation.

BACKGROUND AND OBJECTIVE: It was the aim of this study to compare the sensitivity and specificity of low-dose CT colonography (CTC) with that of optical colonoscopy (OC) in asymptomatic patients undergoing these tests in a screening program for colonic cancer. PATIENTS AND METHODS: 58 patients (mean age 62.6 years) were included. They underwent low dose CTC and, immediately afterwards, colonoscopy. The colonoscopists were unaware of the CTC findings. A "second look" was performed if a lesion seen in CTC had been missed in the first colonoscopy. RESULTS: A total of 150 lesions were detected and histologically confirmed. 136 were found to be polypoid lesions, classified as either hyperplastic polyps (n = 66) or polyps with intraepithelial neoplasia (n = 70). In the per-patient analysis only 22.4 % of patients had no polypoid lesion, 27.6 % had at least one hyperplastic and 50.0 % had at least one adenomatous lesion. Sensitivity for adenomas of all size categories was calculated 55.7 % for CTC and 92.9 % for OC. This marked difference (both for the detection of individual lesions and the per-patient analyses) does not reach significance in the two-sided McNemar test. CONCLUSIONS: There was a high prevalence of lesions with intraepithelial neoplasia in this screening group. OC had a higher sensitivity than CTC in the detection of lesions smaller than 10 mm.

Parallel processing of large datasets from NanoLC-FTICR-MS measurements.

A new approach for automatic parallel processing of large mass spectral datasets in a distributed computing environment is demonstrated to significantly decrease the total processing time. The implementation of this novel approach is described and evaluated for large nanoLC-FTICR-MS datasets. The speed benefits are determined by the network speed and file transfer protocols only and allow almost real-time analysis of complex data (e.g., a 3-gigabyte raw dataset is fully processed within 5 min). Key advantages of this approach are not limited to the improved analysis speed, but also include the improved flexibility, reproducibility, and the possibility to share and reuse the pre- and postprocessing strategies. The storage of all raw data combined with the massively parallel processing approach described here allows the scientist to reprocess data with a different set of parameters (e.g., apodization, calibration, noise reduction), as is recommended by the proteomics community. This approach of parallel processing was developed in the Virtual Laboratory for e-Science (VL-e), a science portal that aims at allowing access to users outside the computer research community. As such, this strategy can be applied to all types of serially acquired large mass spectral datasets such as LC-MS, LC-MS/MS, and high-resolution imaging MS results.

Examples of Fourier transform ion cyclotron resonance mass spectrometry developments: from ion physics to remote access biochemical mass spectrometry.

The application of Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) for high resolution biomolecular analysis has increased greatly after 30 years of innovation since its conception in 1974. FT- ICR-MS can now routinely be used for the analysis of complex organic mixtures such as biological or petrochemical samples. Many of these new possibilities have been the results of many different instrumental developments. This paper provides a mini review of selected instrumental developments that now allow these measurements. The development of soft ionization techniques such as electrospray ionization and matrix assisted laser desorption and ionisation was crucial for the analysis of biological macromolecules. Improved ion transport optics led to an increase in sensitivity. New ICR cell designs complement the capabilities of FT-ICR-MS by allowing a more thorough study of the mechanism and kinetics of ion reactions in the gas-phase. A selected example of electron capture dissociation (ECD) employs these developments to investigate the role of peptide conformation in ECD. Improved electronics and software allow faster and more flexible experiments. All these improvements led to an increase in speed and sensitivity that are necessary to couple FT-MS to fast separation techniques such as nano-high performance liquid chromatography. The modern FT-ICR-MS instruments can be incorporated in virtual organizations allowing remote access to unique infrastructure. This concept of remote experimentation opens new possibilities for scientific collaborations between expert scientists at different locations and allows the efficient use of this expensive instrumentation.

Haloacetates in fog and rain.

Atmospheric haloacetates can arise from photochemical degradation of halogenated hydrocarbons and from direct anthropogenic emissions. Furthermore, there is also evidence of natural sources although these are quantitatively uncertain. As haloacetates are highly soluble in water, hydrometeors are most significant for their deposition. Fogwater (96 samples) and rainwater samples (over 100 samples) were collected from July 1998 to March 1999 at an ecological research site in northeastern Bavaria, Germany. They were analyzed for monofluoroacetate (MFA), difluoroacetate (DFA), trifluoroacetate (TFA), monochloroacetate (MCA), dichloroacetate (DCA), trichloroacetate (TCA), monobromoacetate (MBA), and dibromoacetate (DBA). The major inorganic ions were also determined. High concentrations of up to 11 microg/L MCA, 5 microg/L DCA, 2 microg/L TCA, and 2 microg/L TFA were found in fogwater associated with westerly winds. Backward trajectories were calculated to determine the origin of the air masses. MBA and DBA have highest concentrations in fogwater advected with air originating from the Atlantic, suggesting the marine origin of these two compounds. All analyzed substances show higher average concentrations in fog than in rain. Estimates of the deposition of haloacetates suggest that the contribution of fog may be more important than rain for the total burden of a forest ecosystem.

[Intraoperative transesophageal echocardiography in large heart tumors]. / Intraoperative transösophageale Echokardiographie bei grossen Herztumoren.

A 46-year-old woman known to have an asymptomatic left atrial tumour suddenly developed dyspnoea, tachycardia (up to 140 beats/min) and a fall in systolic blood pressure to 80 mm Hg, 12 days after an extensive radical operation for metastatic ovarian carcinoma. Echocardiography demonstrated a large left atrial tumour which impaired left ventricular filling. A previously healthy 42-year-old man reported increasing exertional dyspnoea with retrosternal feeling of tightness. A 2/6 systolic murmur was audible over the cardiac apex, a 3/6 diastolic murmur heard maximally over the 2nd intercostal space, and there was a third heart sound. The chest X-ray film demonstrated pulmonary venous congestion and there was an enlarged P-wave in the ECG suggesting left-atrial enlargement. Echocardiography revealed a myxoma-like tumour in the left atrium. A thoracotomy was performed in both patients. Using intra-operative transoesophageal echocardiography, the attachment of the tumour (in both instances a myxoma), the course of the operation and, after tumour removal, normal valvar and cardiac function were demonstrated. In case 2, the echocardiographic findings justified an atypical approach via the right atrium. In both patients removal of the tumour and the postoperative course were without complication.