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DOES TRANSMISSION ELECTRON MICROSCOPE PRODUCE 3D IMAGES SERIAL

DOES TRANSMISSION ELECTRON MICROSCOPE PRODUCE 3D IMAGES SERIAL

In our case, a ribbon containing 10-15 serial sections is cut using the diamond knife, and then carefully separated from the knife using an eyelash. The length of the serial-section ribbons is chosen to ensure that they can be picked up using an iron ring with an inner diameter of 7 mm. Ultrathin serial sections (100 nm thick) are cut using an ultramicrotome with a diamond knife.

Preparation and arrangement of serial-section ribbons.

The dimensions of the trapezoid should be trimmed within an upper edge length of 0.7 mm, a lower edge length of 1 mm, and a side height of 0.5 mm. It is important that the upper and lower edges of the trapezoid be trimmed in parallel to ensure that the serial sections are produced with a ribbon-shaped configuration.

8),9)įirst, a resin block is trimmed into a trapezoidal shape using a razor blade. Here, we describe the various steps of serial-section SEM for the Golgi apparatus. Serial-section SEM is a technique in which serial ultrathin sections are mounted on a rigid substrate, and images of these sections are obtained by SEM (Fig. Preparation of samples for serial-section SEM In this report, we provide an introduction to serial-section SEM and discuss its application to the 3D structural analysis of the Golgi apparatus.

In addition, wide areas of the samples can be observed at one time, and any given section can be re-observed any number of times if necessary. First, the complex grid manipulation required for serialsection transmission electron microscopy (TEM) is not required, allowing serial ultrathin sections to be collected with ease. 4-7) This is commonly referred to as serial-section SEM (or array tomography), and it offers a number of advantages over other imaging methods. There have also been reports on 3D reconstruction of SEM images of serial ultrathin sections mounted on a rigid substrate such as glass. Such techniques include FIB/SEM, 1,2) which is a block-face observation method using a focused ion beam (FIB), and serial block-face SEM (SBF/SEM), 3) in which an ultramicrotome is installed inside an SEM system. In recent years, techniques involving three-dimensional (3D) reconstruction of consecutive scanning electron microscopy (SEM) images of resin-embedded specimens have been the subject of increased attention for the structural analysis of biological specimens.