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HItachi S-900 Field-Emission Scanning Electron Microscope

Hitachi S-900 FESEM scanning electron microscope with a cold-cathode field-emission emitter. Using a cold field emission source as the electron emitter instead of tungsten wire or LaB6 crystal produces the brightest beam with the smallest electron source and lowest energy spread. This increases the resolution of the SEM, and allows the use of low accelerating voltages, which helps prevent beam damage and penetration and reduces charging artifacts.

The S-900 also has an Autrata high resolution backscattered detector, which allows imaging of backscattered electrons and much lower accelerating voltages and higher resolution than with a semiconductor or Robinson backscattered electron detector. The Autrata detector produces images of as high a resolution as the secondary detector down to 5kV accelerating voltage.

ImmunoSEM

The Autrata detector makes the S-900 particularly useful for imaging colloidal metals conjugated to antibodies or other ligands attached to cell surface receptors and doing immunoSEM. SEMs are good for immunolabeling of surface structures, as the entire specimen can be examined, instead of simply a narrow slice, as is the usual case with sectioned material. Immunolabeling of internal structures can also be studied in the SEM using colloidal metal conjugates using different techniques to open the specimen to expose the interior. Backscattered electrons can also be used to detect colloidal metals within a sample, as the higher energy of the primary electrons allows them to escape from within the sample. Examples of the use of colloidal gold conjugated antibodies for immunoSEM can be seen on the colloidal metals gallery pages. See the "Colloidal metals and conjugates" page for further information.

Unfortunately, due to the complexity of the instrument, users cannot be trained to operate the S-900 field-emission SEM (FESEM). The microscope must be operated by BBPIC staff.

Please contact Ralph Albrecht or Joe Heintz for further information. All images on this page by Philip Oshel

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Click here for instrument scheduling.

Cell surface distribution of L-selectin adhesion protein on neutrophils. 16 nm gold-conjugated secondary antibody against LAM1-116 against L-selectin, Backscattered electron image using 5kV accelerating voltage
Cell surface distribution of L-selectin adhesion protein on neutrophils. 16 nm gold-conjugated secondary antibody against LAM1-116 against L-selectin, Backscattered electron image using 5kV accelerating voltage. From Doug Steeber, Univ. Wisconsin - Milwaukee, with permission. Venturi, G.M., L. Tu, T. Kadono, A.I. Khan, Y. Fujimoto, P. Oshel, C.B. Brock, A.S. Miller, R.M. Albrecht, P. Kubes, D.A. Steeber and T.F. Tedder. Leukocyte Migration is Regulated by L-Selectin Endoproteolytic Release. Immunity 19:1-20. Click on image for a larger, 380 kB version.
Backscattered electron (BSE) image of isolated mitochondria from cultured bovine lymphocytes. These mitochondria are either in the late stage of dividing, or an early stage of fusing. White dots are 18 nm gold particles conjugated to an antibody against Mannheimia haemolytica leukotoxin. Accelerating voltage was 5kV.
Backscattered electron (BSE) image of isolated mitochondria from cultured bovine lymphocytes. These mitochondria are either in the late stage of dividing, or an early stage of fusing. White dots are 18 nm gold particles conjugated to an antibody against Mannheimia haemolytica leukotoxin. Accelerating voltage was 5kV. Dhammika Atapattu, School of Veterinary Medicine, Univ. of Wisconsin, unpublished with permission. Click on image for a larger, 380 kB version.
Secondary electron (SE) image of the same isolated mitochondria.
Secondary electron (SE) image of the same isolated mitochondria. Accelerating voltage was 5kV. Note how BSE image highlights the colloidal gold immunolabel. Dhammika Atapattu, School of Veterinary Medicine, Univ. of Wisconsin, unpublished with permission. Click on image for a larger, 380 kB version.