Some personal and historical notes on the utility of deep-etch electron microscopy for making cell structure/function correlations

This brief essay talks up the advantages of metal replicas for electron microscopy and explains why they are still the best way to image frozen cells in the electron microscope. Then it explains our approach to freezing, namely the Van Harreveld trick of “slamming” living cells onto a supercold block of metal sprayed with liquid helium at −269ºC, and further talks up this slamming over the alternative of high-pressure freezing, which is much trickier but enjoys greater favor at the moment. This leads me to bemoan the fact that there are not more young investigators today who want to get their hands on electron microscopes and use our approach to get the most “true to life” views of cells out of them with a minimum of hassle. Finally, it ends with a few perspectives on my own career and concludes that, personally, I'm permanently stuck with the view of the “founding fathers” that cell ultrastructure will ultimately display and explain all of cell function, or as Palade said in his Nobel lecture,electron micrographs are “irresistible and half transparent … their meaning buried under only a few years of work,” and “reasonable working hypotheses are already suggested by the ultrastructural organization itself.

Analytical study of the optimum geometric configuration of a space shuttle materials laboratory

A steady state, collisionless flow analysis was made of the density distribution within a hemisphere-disc system due to independent, uniformly distributed internal gas sources. The model was used to estimate the density within a molecular shield, deployed from the shuttle orbiter, which contained internal experiments having a prescribed gas source. Contour plots of the density distribution within the system were presented for disc-to-hemisphere radius ratios of .1, .3, .5, .7, and for disc-to-hemisphere surface emission flux density ratios of .01, 1, 100. The hemisphere-disc system was compared to the empty hemisphere, and it was found that if the disc emission flux density was the same as the hemisphere and the disc radius was not greater than 1/3 of the hemisphere radius, the increase in density at the center of the hemisphere-disc system was less than 50%

The structural basis of long-term potentiation in hippocampal synapses, revealed by electron microscopy imaging of lanthanum-induced synaptic vesicle recycling

Hippocampal neurons in dissociated cell cultures were exposed to the trivalent cation lanthanum for short periods (15-30 min) and prepared for electron microscopy (EM), to evaluate the stimulatory effects of this cation on synaptic ultrastructure. Not only were characteristic ultrastructural changes of exaggerated synaptic vesicle turnover seen within the presynapses of these cultures-including synaptic vesicle depletion and proliferation of vesicle-recycling structures-but the overall architecture of a large proportion of the synapses in the cultures was dramatically altered, due to large postsynaptic bulges or herniations into the presynapses. Moreover, in most cases, these postsynaptic herniations or protrusions produced by lanthanum were seen by EM to distort or break or perforate the so-called postsynaptic densities (PSDs) that harbor receptors and recognition molecules essential for synaptic function. These dramatic EM observations lead us to postulate that such PSD breakages or perforations could very possibly create essential substrates or tags for synaptic growth, simply by creating fragmented free edges around the PSDs, into which new receptors and recognition molecules could be recruited more easily, and thus, they could represent the physical substrate for the important synaptic growth process known as long-term potentiation (LTP). All of this was created simply in hippocampal dissociated cell cultures, and simply by pushing synaptic vesicle recycling way beyond its normal limits with the trivalent cation lanthanum, but we argued in this report that such fundamental changes in synaptic architecture-given that they can occur at all-could also occur at the extremes of normal neuronal activity, which are presumed to lead to learning and memory

An application of failure flow analysis to a GSFC spacecraft project

Application of failure flow analysis to evaluate test program of Explorer 18 satellit

Field-tuned quantum critical point of antiferromagnetic metals

A magnetic field applied to a three-dimensional antiferromagnetic metal can destroy the long-range order and thereby induce a quantum critical point. Such field-induced quantum critical behavior is the focus of many recent experiments. We investigate theoretically the quantum critical behavior of clean antiferromagnetic metals subject to a static, spatially uniform external magnetic field. The external field does not only suppress (or induce in some systems) antiferromagnetism but also influences the dynamics of the order parameter by inducing spin precession. This leads to an exactly marginal correction to spin-fluctuation theory. We investigate how the interplay of precession and damping determines the specific heat, magnetization, magnetocaloric effect, susceptibility and scattering rates. We point out that the precession can change the sign of the leading \sqrt{T} correction to the specific heat coefficient c(T)/T and can induce a characteristic maximum in c(T)/T for certain parameters. We argue that the susceptibility \chi =\partial M/\partial B is the thermodynamic quantity which shows the most significant change upon approaching the quantum critical point and which gives experimental access to the (dangerously irrelevant) spin-spin interactions.Comment: 12 pages, 8 figure

The population of white dwarf binaries with hot subdwarf companions

Hot subdwarfs (sdBs) are core helium-burning stars, which lost almost their entire hydrogen envelope in the red-giant phase. Since a high fraction of those stars are in close binary systems, common envelope ejection is an important formation channel. We identified a total population of 51 close sdB+WD binaries based on time-resolved spectroscopy and multi-band photometry, derive the WD mass distribution and constrain the future evolution of these systems. Most WDs in those binaries have masses significantly below the average mass of single WDs and a high fraction of them might therefore have helium cores. We found 12 systems that will merge in less than a Hubble time and evolve to become either massive C/O WDs, AM\,CVn systems, RCrB stars or even explode as supernovae type Ia.Comment: 5 pages, 2 figures, to appear in the proceedings of the 19th European White Dwarf Workshop, ASP Conf. Se

Eisosome ultrastructure and evolution in fungi, microalgae, and lichens

Eisosomes are among the few remaining eukaryotic cellular differentations that lack a defined function(s). These trough-shaped invaginations of the plasma membrane have largely been studied in Saccharomyces cerevisiae, in which their associated proteins, including two BAR domain proteins, have been identified, and homologues have been found throughout the fungal radiation. Using quick-freeze deep-etch electron microscopy to generate high-resolution replicas of membrane fracture faces without the use of chemical fixation, we report that eisosomes are also present in a subset of red and green microalgae as well as in the cysts of the ciliate Euplotes. Eisosome assembly is closely correlated with both the presence and the nature of cell walls. Microalgal eisosomes vary extensively in topology and internal organization. Unlike fungi, their convex fracture faces can carry lineage-specific arrays of intramembranous particles, and their concave fracture faces usually display fine striations, also seen in fungi, that are pitched at lineage-specific angles and, in some cases, adopt a broad-banded patterning. The conserved genes that encode fungal eisosome-associated proteins are not found in sequenced algal genomes, but we identified genes encoding two algal lineage-specific families of predicted BAR domain proteins, called Green-BAR and Red-BAR, that are candidate eisosome organizers. We propose a model for eisosome formation wherein (i) positively charged recognition patches first establish contact with target membrane regions and (ii) a (partial) unwinding of the coiled-coil conformation of the BAR domains then allows interactions between the hydrophobic faces of their amphipathic helices and the lipid phase of the inner membrane leaflet, generating the striated patterns