scholarly journals Dedicated setup for the photoconversion of fluorescent dyes for functional electron microscopy

2019 ◽  
Author(s):  
Katharine L. Dobson ◽  
Carmel L. Howe ◽  
Yuri Nishimura ◽  
Vincenzo Marra
Keyword(s):  
Electron Microscopy ◽  
Fluorescent Dyes ◽  
Lucifer Yellow ◽  
Light And Electron Microscopy ◽  
Region Of Interest ◽  
Cost Effective ◽  
Cortical Slice ◽  
Time Required ◽  
And Function ◽  
Established Technique

AbstractHere, we describe a cost-effective setup for targeted photoconversion of fluorescent signals into electron dense ones. This approach has offered invaluable insights in the morphology and function of fine neuronal structures. The technique relies on the localized oxidation of diaminobenzidine (DAB) mediated by excited fluorophores. This paper includes a detailed description of how to build a simple photoconversion setup that can increase reliability and throughput of this well-established technique. The system described here, is particularly well-suited for thick neuronal tissue, where light penetration and oxygen diffusion may be limiting DAB oxidation. To demonstrate the system, we use Correlative Light and Electron Microscopy (CLEM) to visualize functionally-labelled individual synaptic vesicles released onto an identified layer 5 neuron in an acute cortical slice. The setup significantly simplifies the photoconversion workflow, increasing the depth of photoillumination, improving the targeting of the region of interest and reducing the time required to process each individual samples. We have tested this setup extensively for the photoconversion of FM 1-43FX and Lucifer Yellow both excited at 473 nm. In principle, the system can be adapted to any dye or nanoparticle able to oxidize DAB when excited by a specific light wavelength.

The Biology of Bothrimonus (=Diplocotyle) (Pseudophyllidea: Cestoda): Detailed Morphology and Fine Structure

1974 ◽  
Vol 31 (2) ◽  
pp. 147-153 ◽  
Author(s):  
M. D. B. Burt ◽  
I. M. Sandeman
Keyword(s):  
Electron Microscopy ◽  
Nervous System ◽  
Fine Structure ◽  
Functional Morphology ◽  
Light And Electron Microscopy ◽  
Nerve Fibers ◽  
Fish Host ◽  
Extensive System ◽  
And Function

Light and electron microscopy were used to describe the functional morphology of Bothrimonus sturionis in detail. In particular, the musculature, nervous system, osmoregulatory system, and tegument are dealt with, and the findings compared with those of other workers. The musculature of the scolex consists of several interrelated systems, the structure of each being discussed in relation to its function. Associated with the regular nervous system, considered typical of cestodes, is an extensive system of giant nerve fibers. The osmoregulatory system is unusual in that there are lateral "excretory" pores in many proglottides which open directly to the exterior of the worm. The microtriches of the tegument are long, like those of other primitive cestodes, and are covered by a noncellular sheath while the worm is in its gammarid host. The sheath is lost when the worm becomes established in its fish host; the nature and function of the sheath are discussed.

Effect of basolateral acidification on the frog oxynticopeptic cell

1990 ◽  
Vol 259 (4) ◽  
pp. G564-G570 ◽  
Author(s):  
S. Arvidsson ◽  
K. Carter ◽  
A. Yanaka ◽  
S. Ito ◽  
W. Silen
Keyword(s):  
Electron Microscopy ◽  
Gastric Mucosa ◽  
Light And Electron Microscopy ◽  
Structure And Function ◽  
Intracellular Acidification ◽  
Intracellular Acidosis ◽  
Normal Morphology ◽  
And Function ◽  
Oxynticopeptic Cells

The effects of intracellular acidosis induced by acidification of the basolateral (nutrient) perfusate on the structure and function of the oxynticopeptic cell were studied in in vitro frog gastric mucosa. Changing the pH of the unbuffered nutrient perfusate (UNB) from 7.2 to 3.5 acidified the oxynticopeptic cell with no change in potential difference (PD) or resistance (R). Intracellular pH (pHi), PD, and R were 7.05 +/- 0.01, 16 +/- 1 mV, 165 +/- 7 omega.cm2 before and 6.44 +/- 0.01, 16 +/- 2 mV, 170 +/- 9 omega.cm2 after nutrient acidification. Acid secretion (H+) increased from 0.86 +/- 0.07 to 1.88 +/- 0.18 mu eq.cm-2.h-1. Addition of forskolin to tissues perfused with nutrient pH (pHn) 3.5 decreased PD to 2 +/- 2 mV and further increased H+ to 3.07 +/- 0.19 mu eq.cm-2.h-1. By light and electron microscopy oxynticopeptic cells perfused with UNB, pHn 3.5, appeared normal. Oxynticopeptic cells in tissues pretreated with omeprazole and then exposed to UNB, pHn 3.5, had extensive morphological damage. On increasing the pH of the nutrient perfusate from 3.5 to 7.2 there was prompt recovery of pHi in untreated and forskolin-stimulated mucosae (pHi 6.87 +/- 0.06 and 6.85 +/- 0.04) but no recovery of pHi in tissues pretreated with omeprazole or cimetidine (pHi 6.26 +/- 0.04 and 6.44 +/- 0.06, n = 6, 30 min after reexposure to UNB, pHn 7.2). We conclude that in a secreting mucosa intracellular acidification of the oxynticopeptic cell to pHi 6.4 is associated with normal morphology, PD, R, and increased H+, and that intracellular acidosis is not de facto deleterious.

scholarly journals FORMATION OF BONE TISSUE IN CULTURE FROM ISOLATED BONE CELLS

1974 ◽  
Vol 61 (2) ◽  
pp. 427-439 ◽  
Author(s):  
Itzhak Binderman ◽  
Dan Duksin ◽  
Arieh Harell ◽  
Ephraim Katzir (Katchalski) ◽  
Leo Sachs
Keyword(s):  
Electron Microscopy ◽  
Bone Tissue ◽  
Bone Cells ◽  
Light And Electron Microscopy ◽  
Bone Collagen ◽  
The Matrix ◽  
Three Stages ◽  
And Function

A system is described for the formation of bone tissue in culture from isolated rat bone cells. The isolated bone cells were obtained from embryonic rat calvarium and periosteum or from traumatized, lifted periosteum of young rats. The cells were cultured for a period of up to 8 wk, during which time the morphological, biochemical, and functional properties of the cultures were studied. Formation of bone tissue by these isolated bone cells was shown, in that the cells demonstrated osteoblastic morphology in light and electron microscopy, the collagen formed was similar to bone collagen, there was mineralization specific for bone, and the cells reacted to the hormone calcitonin by increased calcium ion uptake. Calcification of the fine structure of the cells and the matrix is described. Three stages in the calcification process were observed by electron microscopy. It is concluded that these bone cells growing in vitro are able to function in a way similar to such cells in vivo. This tissue culture system starting from isolated bone cells is therefore suitable for studies on the structure and function of bone.

scholarly journals Presence of adenosine deaminase on the surface of mononuclear blood cells: immunochemical localization using light and electron microscopy.

1991 ◽  
Vol 39 (8) ◽  
pp. 1001-1008 ◽  
Author(s):  
J M Aran ◽  
D Colomer ◽  
E Matutes ◽  
J L Vives-Corrons ◽  
R Franco
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Adenosine Deaminase ◽  
Blood Cells ◽  
Light And Electron Microscopy ◽  
Enzyme Expression ◽  
And Function ◽  
Immune Dysfunctions ◽  
Mononuclear Blood Cells ◽  
Immunodeficiency Syndromes

Adenosine deaminase, which is essential for lymphoid differentiation and function, has previously been considered to be a cytosolic enzyme. In this report we demonstrate that it can be found associated with the plasma membrane of lymphocytes. By means of immunological techniques using both light and electron microscopy, adenosine deaminase was localized on the external side of the plasma membrane of normal lymphocytes and monocytes. Since the enzyme expression differed depending on the type of cell examined, new hypotheses about the mechanisms involving purine metabolism in immune dysfunctions or immunodeficiency syndromes may be considered.

scholarly journals In situ strategy for biomedical target localization via nanogold nucleation and secondary growth

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Akira Sawaguchi ◽  
Takeshi Kamimura ◽  
Nobuyasu Takahashi ◽  
Atsushi Yamashita ◽  
Yujiro Asada ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Self Assembly ◽  
Light And Electron Microscopy ◽  
Secondary Growth ◽  
Spatial Location ◽  
Tissue Samples ◽  
Cell Tissue ◽  
Nanogold Particles ◽  
And Function

AbstractImmunocytochemistry visualizes the exact spatial location of target molecules. The most common strategy for ultrastructural immunocytochemistry is the conjugation of nanogold particles to antibodies as probes. However, conventional nanogold labelling requires time-consuming nanogold probe preparation and ultrathin sectioning of cell/tissue samples. Here, we introduce an in situ strategy involving nanogold nucleation in immunoenzymatic products on universal paraffin/cryostat sections and provide unique insight into nanogold development under hot-humid air conditions. Nanogold particles were specifically localized on kidney podocytes to target synaptopodin. Transmission electron microscopy revealed secondary growth and self-assembly that could be experimentally controlled by bovine serum albumin stabilization and phosphate-buffered saline acceleration. Valuable retrospective nanogold labelling for gastric H+/K+-ATPase was achieved on vintage immunoenzymatic deposits after a long lapse of 15 years (i.e., 15-year-old deposits). The present in situ nanogold labelling is anticipated to fill the gap between light and electron microscopy to correlate cell/tissue structure and function.

Blood Supply and Function of the Tegmentum Vasculosum (Stria Vascularis) of the Duckling Cochlea

1999 ◽  
Vol 5 (S2) ◽  
pp. 1194-1195
Author(s):  
F.E. Hossler
Keyword(s):  
Electron Microscopy ◽  
Blood Supply ◽  
Carotid Arteries ◽  
Stria Vascularis ◽  
Light And Electron Microscopy ◽  
Corrosion Casting ◽  
The Common ◽  
And Function ◽  
Tegmentum Vasculosum ◽  
Common Carotid Arteries

On the basis of ultrastructural, biochemical, cytochemical, and electro-physiological evidence, the stria vascularis, a highly vascular epithelium of the mammalian cochlea, has been implicated as one site of endolymph synthesis. It would seem natural to suspect that its highly developed counterpart in the avian cochlea, the tegmentum vasculosum (TV), might have a similar function. This study utilizes routine light and electron microscopy, biochemistry, cytochemistry, and vascular corrosion casting to describe the structure and blood supply to the (TV) in the duckling and provide evidence for its involvement in endolymph synthesis.Four to 16 week-old ducklings (Anas platyrhyncos)were anesthetized and anticoagulated and perfused via the common carotid arteries with buffered, 2% glutaraldehyde for routine light microscopy or transmission or scanning electron microscopy, or with Mercox resin for vascular corrosion casting. Casts were macerated in 5% KOH and warm water to remove tissue, cleaned in formic acid and water, dried by lyophilization, mounted on stubs and viewed at 10-20 kv by SEM.

Microscopy, Electron Microscopy, Elemental Analysis, Image Analysis, Digital Photography, And Telepathology Applications In Pharmaceuticals Research

1999 ◽  
Vol 5 (S2) ◽  
pp. 6-7
Author(s):  
G. Argentieri ◽  
P. Grosenstein ◽  
K. Killary
Keyword(s):  
Electron Microscopy ◽  
Image Analysis ◽  
Quality Assurance ◽  
Drug Discovery ◽  
Research And Development ◽  
Light And Electron Microscopy ◽  
Pharmaceutical Research ◽  
Cost Effective ◽  
Wide Range ◽  
Functional Areas

Light and electron microscopy are exceedingly versatile disciplines having many applications in pharmaceutical research. Many corporate and university facilities are organized with a centralized microscopy laboratory that serves the needs of many different investigators. This is also true at the Novartis Pharmaceuticals Corporation in East Hanover, New Jersey. A centralized microscopy laboratory is not only more efficient, but also most cost effective. The microscopy lab at Novartis is designed to meet the needs for a wide range of applications serving all of Research and Development. The lab contains seven functional areas for microscopy applications. A large preparation and processing area, necropsy, microtomy, TEM, SEM, wet and dry dark rooms, and an image analysis suite including telepathology, complete the lab.Microscopy service is widely used in Pathology, Toxicology, Drug Discovery, Technical Research and Development, Production and Quality Assurance, and Compliance. Microscopy services have also been involved in marketing (packaging labels and commercials) for Novartis products.

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