Low-Voltage Scanning Electron Microscopy of Mammalian Fertilization In Vitro: Preparation of Oocytes

1997 ◽  
Vol 3 (3) ◽  
pp. 193-202 ◽  
Author(s):  
Mark W. Tengowski ◽  
Gerald Schatten
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
High Pressure ◽  
Low Voltage ◽  
Fertilization In Vitro ◽  
Aldehyde Fixation ◽  
Bovine Oocytes ◽  
Voltage Scanning ◽  
Scanning Electron

Abstract: To optimize specimen-processing protocols for investigations of fertilization in mammals, with high-resolution, low-voltage scanning electron microscopy (LVSEM), bovine oocytes matured in vitro were prepared by either aldehyde fixation, conductive staining, or high-pressure cryoimmobilization with freeze-substitution. Samples prepared by these different preparative techniques were coated with either platinum or gold and imaged with an LVSEM operating at 1.5 keV. Additionally, aldehyde-fixed oocytes, sputter-coated with gold, were imaged with a conventional scanning electron microscope (cSEM). The results show that bovine oocytes prepared by aldehyde fixation or cryoimmobilized by high-pressure freezing produced superior images of the zona pellucida (ZP) compared with those from the conductive stained samples. A comparison of LVSEM and cSEM images suggests that the gold-coating employed in cSEM preparation obscures the ZP detail seen with the LVSEM, but not with the cSEM. Application of these procedures provide not only a new view of the ZP but may be useful in elucidating the molecular mechanism of gamete interactions during the fertilization process.

Novel Approaches to Low-Voltage Scanning Electron Microscopy

1990 ◽  
Vol 48 (1) ◽  
pp. 366-367
Author(s):  
Arthur V. Jones
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Low Voltage ◽  
Electron Optics ◽  
Current Interest ◽  
New Concepts ◽  
Opportune Time ◽  
Novel Approaches ◽  
Voltage Scanning ◽  
Scanning Electron

In comparison with the developers of other forms of instrumentation, scanning electron microscope manufacturers are among the most conservative of people. New concepts usually must wait many years before being exploited commercially. The field emission gun, developed by Albert Crewe and his coworkers in 1968 is only now becoming widely available in commercial instruments, while the innovative lens designs of Mulvey are still waiting to be commercially exploited. The associated electronics is still in general based on operating procedures which have changed little since the original microscopes of Oatley and his co-workers.The current interest in low-voltage scanning electron microscopy will, if sub-nanometer resolution is to be obtained in a useable instrument, lead to fundamental changes in the design of the electron optics. Perhaps this is an opportune time to consider other fundamental changes in scanning electron microscopy instrumentation.

scholarly journals Low Voltage Scanning Electron Microscopy

2002 ◽  
Vol 10 (2) ◽  
pp. 22-23 ◽  
Author(s):  
David C Joy ◽  
Dale E Newbury
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
High Spatial Resolution ◽  
Low Voltage ◽  
Incident Beam ◽  
Operational Mode ◽  
X Ray ◽  
Rapid Fall ◽  
Voltage Scanning ◽  
Scanning Electron

Low Voltage Scanning Electron Microscopy (LVSEM), defined as operation in the energy range below 5 keV, has become perhaps the most important single operational mode of the SEM. This is because the LVSEM offers advantages in the imaging of surfaces, in the observation of poorly conducting and insulating materials, and for high spatial resolution X-ray microanalysis. These benefits all occur because a reduction in the energy Eo of the incident beam leads to a rapid fall in the range R of the electrons since R ∼k.E01.66. The reduction in the penetration of the beam has important consequences.

Low voltage scanning electron microscopy

Micron ◽  
1996 ◽  
Vol 27 (3-4) ◽  
pp. 247-263 ◽  
Author(s):  
David C. Joy ◽  
Carolyn S. Joy
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Low Voltage ◽  
Voltage Scanning ◽  
Scanning Electron

On Low Voltage Scanning Electron Microscopy and Chemical Microanalysis

2000 ◽  
Vol 6 (4) ◽  
pp. 307-316 ◽  
Author(s):  
E.D. Boyes
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Spatial Resolution ◽  
Low Voltage ◽  
Energy Dispersive ◽  
Beam Specimen ◽  
Topographic Analysis ◽  
X Ray ◽  
Voltage Scanning ◽  
Scanning Electron

AbstractThe current status and general applicability of scanning electron microscopy (SEM) at low voltages is reviewed for both imaging (low voltage scanning electron microscopy, LVSEM) and chemical microanalysis (low voltage energy-dispersive X-ray spectrometry, LVEDX). With improved instrument performance low beam energies continue to have the expected advantages for the secondary electron imaging of low atomic number (Z) and electrically non-conducting samples. They also provide general improvements in the veracity of surface topographic analysis with conducting samples of all Z and at both low and high magnifications. In new experiments the backscattered electron (BSE) signal retains monotonic Z dependence to low voltages (<1 kV). This is contrary to long standing results in the prior literature and opens up fast chemical mapping with low dose and very high (nm-scale) spatial resolution. Similarly, energy-dispersive X-ray chemical microanalysis of bulk samples is extended to submicron, and in some cases to <0.1 μm, spatial resolution in three dimensions at voltages <5 kV. In favorable cases, such as the analysis of carbon overlayers at 1.5 kV, the thickness sensitivity for surface layers is extended to <2 nm, but the integrity of the sample surface is then of concern. At low beam energies (E0) the penetration range into the sample, and hence the X-ray escape path length out of it, is systematically restricted (R = F(E05/3)), with advantages for the accuracy or elimination of complex analysis-by-analysis matrix corrections for absorption (A) and fluorescence (F). The Z terms become more sensitive to E0 but they require only one-time calibrations for each element. The new approach is to make the physics of the beam–specimen interactions the primary factor and to design enabling instrumentation accordingly.

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