Wareham Sc Systems Inc., Cambridge, MA, USA) for complete micro-TEM of tissues. When tested using an inverted microscope and a Leica-Emission Laser Scattering electron microscope (LM-FV1200; Leica Microsystems; Wetzlar, Germany), small blood vessels were observed through scanning electron microscopy and are shown in lower panel of **[Fig. S1](#pone.0089856.s001){ref-type=”supplementary-material”}**. The electron optics arrangement of hematoxylin-eosin (H&E) stroma is shown in lower panel of **[Fig. S2](#pone.
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0089856.s002){ref-type=”supplementary-material”}**. LSM staining was used for determining the location and distribution in HeLa (lower panel) and human osteosarcoma (top panel). Scanning electron microscope (SEM) images were taken with a 200 mesh charge grid. More images were taken with 6 MZYX plates (50 fb), which allowed electron microscopy to be conducted simultaneously with cell phone counting under a microscope. Differentiation of Tissue by Matrigel {#s2j} ———————————— Tumor tissue was diluted with Tris-buffered saline (TBS) after centrifugation for 10 min at 4°C. After 0.5 h, the tissue was carefully transferred to a polytetrafluoroethylene (PTFE) filter and the sample (10 mg) was shaken for 6 min at room temperature in a high-power hood, frozen in −80°C for 20 min, and stored at −80°C until further processing.
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Matrigel medium (Millipore Sigma; Menomun Bioscience, Milan, Italy) was substituted for the PTFE filter for one technical replicate. This new medium was stored on a special freezer block in the S. Pont-Mercier de Paulique, France on 1 guage, 2 guage and 2 guage. Results and Discussion {#s3} ====================== Our studies were set in temporal and spatial phase in the cortex, and evaluated the degree of co-localization and internalization with two main metrics. The use of real in vivo imaging with the z-bio MINT beam scanner gives a good guide for examining the spatio-temporal relationships between cortical regions and the surrounding tissue when using only in situ inactivation techniques. Our new MINT based on atomic force microscopy in the z-direction with an instrument called S. Pont-Mercier is more sensitive than S. Pont-Merle for high resolution imaging of cortical tissues.
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The S. Pont-Mercier instrument measures the modality transfer of the z-plane for 1.5 Hz exposure mode. The z-transposition is not required under some experimental conditions, including those exposed to laser diffraction in the microscope ([@pone.0089856-Lucic1], [@pone.0089856-Lucic2], [@pone.0089856-Lucic3], [@pone.0089856-Pekka1].
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We measured three different spatial distributions (**[Fig. 1A](#pone-0089856-g001){ref-type=”fig”}**) in the primary cortex, the cerebellum, and the temporal cortex. The first and second quantities range between 32 and -8.5°, and 16°, respectively, in the cerebellum. The only significant difference between the two regions was between the cerebellum and temporal cortex. Quantification was performed by using three different algorithms, which were applied see this determine relative changes in the corresponding pixels of the z-regions of the tissue. Scaling of the intensity was used according to the TPS, in different units of the MINT imaging device; we found that the relative changes were in lower intensity units (30%, median) at both anterior and posterior segments. 








