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For the localization of N in the specimens, the electron spectroscopic images were taken at ΔE = 410 eV, just above the ionization edge (IE) of N (Nk ΔE = 401 eV), to detect the total N signal, and at ΔE = 377 eV, the pre-ionization edge (PIE) of N, as a reference carrying information on the background [39], [40].
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As its dichroic-coated glass fins shimmer with rainbow iridescence in the late summer sun, it could be one of the specimens under the electron microscope buried in the bowels of this new £700m biomedical research facility.
Elaborate data acquisition schemes with autotuning functions minimize exposure of the specimen to the electron beam and sophisticated image analysis routines retrieve a maximum of information from noisy data sets.
Here, we report the development of methods to account for local variations in defocus and beam-induced drift, and the implementation of a data-driven dose compensation scheme that significantly improves the extraction of high-resolution information recorded during exposure of the specimen to the electron beam.
A major factor limiting resolution in cryo-electron tomography is radiation damage of the specimen by the electron beam during acquisition of a tilt series.
Carbon film can reduce the beam-induced charging and movement of the specimen in the electron microscope and it is commonly used as a support for 2D crystals [ 48].
In addition, any specimens which at the electron microscope level, showed faulty sampling or preparation were discarded, leaving us with 16 carcinoma and 15 normal specimens for detailed study.
A 3D reconstruction at 25 Å resolution of native hemoglobin of the polychaete worm Arenicola marina was carried out from frozen-hydrated specimens examined in the electron microscope.
Nevertheless, a residual background gradient remained in the reconstructed phase images, which might stem from specimen charging under the electron beam and therefore may affect (V_{text {Fields}}).
The cumulative dose of electrons for the entire series of 60 70 images was thus within acceptable limits of 5000 electrons/nm2 of the specimen (that included the electron dose spent on searching for suitable areas and for area tracking during data collection).
A morphologic diagnosis may be obtained within 10 min of specimen arrival in the electron microscopic facility.
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