Structural Investigation of Invertebrate Hemoglobins Using the Stem

1980 ◽  
Vol 38 ◽  
pp. 676-677
Author(s):  
O.H. Kapp ◽  
M. Ohtsuki ◽  
S.N. Vinogradov
Keyword(s):  
Scanning Transmission Electron Microscopy ◽  
Structural Investigation ◽  
Uranyl Acetate ◽  
Carbon Substrate ◽  
Distilled Water ◽  
Acetate Solution ◽  
Sedimentation Constant ◽  
Transmission Electron ◽  
Scanning Transmission ◽  
Polypeptide Chains

Scanning transmission electron microscopy (STEM) was used to study the structure of the extracellular hemoglobins of the annelids Lumbricus, Arenicola, and Nephtys. These are giant molecules, possessing a molecular weight of 3-4 x IO6,a sedimentation constant of about 60s and consisting of about five to seven different polypeptide chains (1,2,3). The samples were diluted to 100-200μg/ml with distilled water just before application to a very thin(∽15Å) carbon substrate supported on a microgrid. One percent (w/v) uranyl acetate solution was used for negative staining for 2 min. and dried in air. The specimens were examined with an electron energy of 22-35keV.

Scanning Transmission Electron Microscopy (STEM) of Annelid Extracellular Hemoglobins

1981 ◽  
Vol 39 ◽  
pp. 432-433
Author(s):  
O. H. Kapp ◽  
M. Ohtsuki ◽  
S. N. Vinogradov ◽  
A. V. Crewe
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Scanning Transmission Electron Microscopy ◽  
Lumbricus Terrestris ◽  
Carbon Substrate ◽  
Acetate Solution ◽  
Scanning Transmission Electron ◽  
Transmission Electron ◽  
Scanning Transmission ◽  
On Line

Scanning transmission electron microscopy (STEM) was used to study the structure of the extracellular hemoglobin of the annelid Lumbricus terrestris. These are giant molecules possessing a molecular weight of approximately 3.9 x 106 and appear in electron micrographs as a hexagonal bilayer, The samples for this study were diluted to 100 μg/ml with distilled water and applied to a thin ( ∽15 Å) carbon substrate supported on a microgrid. One percent (w/v) uranyl acetate solution was used for negative staining for 2 minutes and dried in air. STEM images were recorded using both a high resolution CRT with 1024 x 1024 picture elements and an on-line computer recording with 512 x 512 pixels. The images were rotated in on-line fashion with the computer to enhance contrast.The hemoglobin of Lumbricus assumes three common positions on the substrate (Fig. 1). The top view (Fig. la) demonstrates that the 1/12th subunits possess an approximately pentagonal shape with a central region of low protein density.

High-Voltage Scanning Electron Microscopy

1970 ◽  
Vol 28 ◽  
pp. 6-7
Author(s):  
J. M. Cowley
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Scanning Transmission Electron Microscopy ◽  
Wave Optics ◽  
Contrast Effects ◽  
Transmission Electron ◽  
Mode Of Operation ◽  
Scanning Transmission ◽  
Types Of Information ◽  
Voltage Scanning

The comparison of scanning transmission electron microscopy (STEM) with conventional transmission electron microscopy (CTEM) can best be made by means of the Reciprocity Theorem of wave optics. In Fig. 1 the intensity measured at a point A’ in the CTEM image due to emission from a point B’ in the electron source is equated to the intensity at a point of the detector, B, due to emission from a point A In the source In the STEM. On this basis it can be demonstrated that contrast effects In the two types of instrument will be similar. The reciprocity relationship can be carried further to include the Instrument design and experimental procedures required to obtain particular types of information. For any. mode of operation providing particular information with one type of microscope, the analagous type of operation giving the same information can be postulated for the other type of microscope. Then the choice between the two types of instrument depends on the practical convenience for obtaining the required Information.

Reconstruction of Two-Dimensional Projections of GP 32*I

1981 ◽  
Vol 39 ◽  
pp. 38-39
Author(s):  
H.A. Cohen ◽  
W. Chiu ◽  
J. Hosoda
Keyword(s):  
Electron Microscopy ◽  
Molecular Weight ◽  
Uranyl Acetate ◽  
Distilled Water ◽  
Acetate Solution ◽  
Two Dimensional ◽  
Parent Molecule ◽  
T4 Bacteriophage ◽  
Electron Imaging ◽  
Better Than

GP 32 (molecular weight 35000) is a T4 bacteriophage protein that destabilizes the DNA helix. The fragment GP32*I (77% of the total weight), which destabilizes helices better than does the parent molecule, crystallizes as platelets thin enough for electron diffraction and electron imaging. In this paper we discuss the structure of this protein as revealed in images reconstructed from stained and unstained crystals.Crystals were prepared as previously described. Crystals for electron microscopy were pelleted from the buffer suspension, washed in distilled water, and resuspended in 1% glucose. Two lambda droplets were placed on grids over freshly evaporated carbon, allowed to sit for five minutes, and then were drained. Stained crystals were prepared the same way, except that prior to draining the droplet, two lambda of aqueous 1% uranyl acetate solution were applied for 20 seconds. Micrographs were produced using less than 2 e/Å2 for unstained crystals or less than 8 e/Å2 for stained crystals.

Imaging and diffraction modes in Scanning Transmission Electron Microscopy

1986 ◽  
Vol 44 ◽  
pp. 684-687
Author(s):  
J. M. Cowley ◽  
R. Glaisher ◽  
J. A. Lin ◽  
H.-J. Ou
Keyword(s):  
Scanning Transmission Electron Microscopy ◽  
High Efficiency ◽  
Fiber Optic ◽  
Image Intensifier ◽  
Detector System ◽  
Detector Plane ◽  
Secondary Electrons ◽  
Transmission Electron ◽  
Scanning Transmission ◽  
Special Detector

Some of the most important applications of STEM depend on the variety of imaging and diffraction made possible by the versatility of the detector system and the serial nature, of the image acquisition. A special detector system, previously described, has been added to our STEM instrument to allow us to take full advantage of this versatility. In this, the diffraction pattern in the detector plane may be formed on either of two phosphor screens, one with P47 (very fast) phosphor and the other with P20 (high efficiency) phosphor. The light from the phosphor is conveyed through a fiber-optic rod to an image intensifier and TV system and may be photographed, recorded on videotape, or stored digitally on a frame store. The P47 screen has a hole through it to allow electrons to enter a Gatan EELS spectrometer. Recently a modified SEM detector has been added so that high resolution (10Å) imaging with secondary electrons may be used in conjunction with other modes.

A combined pseudoreplica-immunocytochemical technique for research and diagnostic virology

1990 ◽  
Vol 48 (3) ◽  
pp. 900-901
Author(s):  
Blayne Fritz ◽  
Stanley J. Naides ◽  
Kenneth Moore
Keyword(s):  
Phosphotungstic Acid ◽  
Immunogold Labelling ◽  
Uranyl Acetate ◽  
Distilled Water ◽  
Clinical Specimens ◽  
Tissue Sections ◽  
Specimen Retrieval ◽  
Transmission Electron ◽  
Viral Particles ◽  
Diagnostic Virology

The pseudoreplica method of staining viral particles for visualization by transmission electron microscopy is a very popular technique. The ability to concentrate clinical specimens while semi-embedding viral particles makes it especially well suited for morphologic and diagnostic virology. Immunolabelling viral particles with colloidal gold is a technique frequently employed by both research and diagnostic virologists. We have characterized a procedure which provides the advantage of both by modifying and combining pseudoreplica staining and immunogold labelling.Modification of specimen retrieval and delay of staining allows us to utilize pseudoreplica processed specimens within our standard immunogold labelling protocol. In brief, we absorbed samples onto 2% agarose, added.25% Formvar and wicked dry. We then floated the Formvar-virus film onto double distilled water, added grids and retrieved with parafilm. The Formvar-virus specimens were then treated as thin tissue sections within our standard two stage immunolabelling protocol. Following completion of immunogold labelling; each grid was negatively stained with phosphotungstic acid or uranyl acetate contrast stains.

Scanning Transmission Electron Microscopy of Polyethylene Crystals

1980 ◽  
Vol 38 ◽  
pp. 242-245
Author(s):  
F. Khoury ◽  
L. H. Bolz
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Crystallization Temperature ◽  
Scanning Transmission Electron Microscopy ◽  
Growth Habit ◽  
Lateral Growth ◽  
Scanning Transmission Electron ◽  
Transmission Electron ◽  
Scanning Transmission ◽  
Growth Habits

The lateral growth habits and non-planar conformations of polyethylene crystals grown from dilute solutions (<0.1% wt./vol.) are known to vary depending on the crystallization temperature.1-3 With the notable exception of a study by Keith2, most previous studies have been limited to crystals grown at <95°C. The trend in the change of the lateral growth habit of the crystals with increasing crystallization temperature (other factors remaining equal, i.e. polymer mol. wt. and concentration, solvent) is illustrated in Fig.l. The lateral growth faces in the lozenge shaped type of crystal (Fig.la) which is formed at lower temperatures are {110}. Crystals formed at higher temperatures exhibit 'truncated' profiles (Figs. lb,c) and are bound laterally by (110) and (200} growth faces. In addition, the shape of the latter crystals is all the more truncated (Fig.lc), and hence all the more elongated parallel to the b-axis, the higher the crystallization temperature.

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