New Methods for the Generation of Carbohydrate Arrays on Glass Slides and Their Evaluation

ChemBioChem ◽  
2005 ◽  
Vol 6 (6) ◽  
pp. 1007-1015 ◽  
Author(s):  
Moritz B. Biskup ◽  
Jan U. Müller ◽  
Ralf Weingart ◽  
Richard R. Schmidt
Keyword(s):  
New Methods ◽  
Glass Slides ◽  
Carbohydrate Arrays

Thiol–ene immobilisation of carbohydrates onto glass slides as a simple alternative to gold–thiol monolayers, amines or lipid binding

2015 ◽  
Vol 3 (1) ◽  
pp. 175-181 ◽  
Author(s):  
Caroline I. Biggs ◽  
Steve Edmondson ◽  
Matthew I. Gibson
Keyword(s):  
Lipid Binding ◽  
Glass Slides ◽  
Thiol Monolayers ◽  
Simple Alternative ◽  
Carbohydrate Arrays ◽  
New Treatments

Carbohydrate arrays are a vital tool in studying infection, probing the mechanisms of bacterial, viral and toxin adhesion and the development of new treatments, by mimicking the structure of the glycocalyx.

Replication of Wet Objects and Cells

1974 ◽  
Vol 32 ◽  
pp. 102-103
Author(s):  
S. Basu ◽  
D. F. Parsons
Keyword(s):  
Water Vapor Pressure ◽  
High Vacuum ◽  
Polystyrene Latex ◽  
Vacuum System ◽  
Saturated Water Vapor ◽  
Replication Method ◽  
New Methods ◽  
Saturated Water ◽  
Water Vapors ◽  
Intermediate Chamber

We are approaching the invasiveness of cancer cells from the studies of their wet surface morphology which should distinguish them from their normal counterparts. In this report attempts have been made to provide physical basis and background work to a wet replication method with a differentially pumped hydration chamber (Fig. 1) (1,2), to apply this knowledge for obtaining replica of some specimens of known features (e.g. polystyrene latex) and finally to realize more specific problems and to improvize new methods and instrumentation for their rectification. In principle, the evaporant molecules penetrate through a pair of apertures (250, 350μ), through water vapors and is, then, deposited on the specimen. An intermediate chamber between the apertures is pumped independently of the high vacuum system. The size of the apertures is sufficiently small so that full saturated water vapor pressure is maintained near the specimen.

A Freeze Shadow Technique for the Preparation of Soft Paint Latexes for Electron Microscopy

1977 ◽  
Vol 35 ◽  
pp. 314-315
Author(s):  
Earl R. Walter ◽  
Glen H. Bryant
Keyword(s):  
Sample Preparation ◽  
High Vacuum ◽  
Vacuum System ◽  
Latex Particles ◽  
New Methods ◽  
Diffusion Pump ◽  
Film Forming ◽  
Routine Basis ◽  
Distribution Studies ◽  
Preparation Techniques

With the development of soft, film forming latexes for use in paints and other coatings applications, it became desirable to develop new methods of sample preparation for latex particle size distribution studies with the electron microscope. Conventional latex sample preparation techniques were inadequate due to the pronounced tendency of these new soft latex particles to distort, flatten and fuse on the substrate when they dried. In order to avoid these complications and obtain electron micrographs of undistorted latex particles of soft resins, a freeze-dry, cold shadowing technique was developed. The method has now been used in our laboratory on a routine basis for several years.The cold shadowing is done in a specially constructed vacuum system, having a conventional mechanical fore pump and oil diffusion pump supplying vacuum. The system incorporates bellows type high vacuum valves to permit a prepump cycle and opening of the shadowing chamber without shutting down the oil diffusion pump. A baffeled sorption trap isolates the shadowing chamber from the pumps.

Processing Immunoperoxidase Labeled Cell Monolayers and Cryostat Sesctions For Electron Microscopy

1985 ◽  
Vol 43 ◽  
pp. 714-715
Author(s):  
K. Chien ◽  
R. Van de Velde ◽  
I.P. Shintaku ◽  
A.F. Sassoon
Keyword(s):  
Cell Monolayer ◽  
Cell Types ◽  
Surface Antigens ◽  
Immunoperoxidase Method ◽  
Reaction Step ◽  
Hot Plate ◽  
Air Drying ◽  
New Approach ◽  
Cell Monolayers ◽  
Glass Slides

Immunoelectron microscopy of neoplastic lymphoma cells is valuable for precise localization of surface antigens and identification of cell types. We have developed a new approach in which the immunohistochemical staining can be evaluated prior to embedding for EM and desired area subsequently selected for ultrathin sectioning.A freshly prepared lymphoma cell suspension is spun onto polylysine hydrobromide- coated glass slides by cytocentrifugation and immediately fixed without air drying in polylysine paraformaldehyde (PLP) fixative. After rinsing in PBS, slides are stained by a 3-step immunoperoxidase method. Cell monolayer is then fixed in buffered 3% glutaraldehyde prior to DAB reaction. After the DAB reaction step, wet monolayers can be examined under LM for presence of brown reaction product and selected monolayers then processed by routine methods for EM and embedded with the Chien Re-embedding Mold. After the polymerization, the epoxy blocks are easily separated from the glass slides by heatingon a 100°C hot plate for 20 seconds.

Directional observation on lipid of male nectary of Lindera megaphylla hemsl. by the rapid embedding method with polyethylene glycol (PEG)

1990 ◽  
Vol 48 (3) ◽  
pp. 718-719
Author(s):  
Dai Dalin ◽  
Guo Jianmin
Keyword(s):  
Electron Microscope ◽  
Polyethylene Glycol ◽  
Chemical Reaction ◽  
Traditional Method ◽  
Osmium Tetroxide ◽  
Unsaturated Lipid ◽  
Embedding Method ◽  
Long Period ◽  
New Methods

Lipid cytochemistry has not yet advanced far at the EM level. A major problem has been the loss of lipid during dehydration and embedding. Although the adoption of glutaraldehyde and osmium tetroxide accelerate the chemical reaction of lipid and osmium tetroxide can react on the double bouds of unsaturated lipid to from the osmium black, osmium tetroxide can be reduced in saturated lipid and subsequently some of unsaturated lipid are lost during dehydration. In order to reduce the loss of lipid by traditional method, some researchers adopted a few new methods, such as the change of embedding procedure and the adoption of new embedding media, to solve the problem. In a sense, these new methods are effective. They, however, usually require a long period of preparation. In this paper, we do research on the fiora nectary strucure of lauraceae by the rapid-embedding method wwith PEG under electron microscope and attempt to find a better method to solve the problem mentioned above.

Detection of atrial natriuretic peptides (ANP) in rat atria by immunogold-silver staining (IGSS)

1994 ◽  
Vol 52 ◽  
pp. 304-305
Author(s):  
Robyn Rufner ◽  
Gerhard W. Hacker ◽  
Michele Forte ◽  
Nancyleigh E. Carson ◽  
Cristina Xenachis ◽  
...  
Keyword(s):  
Natriuretic Peptides ◽  
Silver Staining ◽  
Sodium Thiosulfate ◽  
Atrial Natriuretic Peptides ◽  
Paraffin Sections ◽  
Glass Slides ◽  
Metallic Salts ◽  
Wistar Kyoto ◽  
Right Atria

The use of immunogold-silver staining (IGSS) to enhance label penetration and Localization for immunocytochemistry or in situ hybridization utilizing a variety of metallic salts has been documented. In this morphological study, the effects of silver acetate, silver lactate and silver nitrate were evaluated for immunogold-labeling of a trial natriuretic peptides (ANP) in rat right atria.Eight Wistar Kyoto retired breeders were sedated with pentobarbital, perfused with either 4% paraformaldehyde (LM) or Karnovsky's fixative (EM), and right atria were dissected, processed, embedded in paraffin or epon, respectively and sectioned according to conventional methods. For light microscopy, an indirect IGSS method according to Hacker (3) was performed. Paraffin sections on glass slides were washed in ddH2O, immersed in Lugol's iodine, washed in ddH2O and treated with 2.5% aqueous sodium thiosulfate for 20 sec. After additional washes in ddH2O and TBS-0.1% fish gelatin, 10% normal goat serum (PBS with 1% BSA) was applied for 20 min before an overnight incubation at 4°C with a polyclonal α-ANP primary antibody (Peninsula Labs, 1:1000 in TBS/BSA).

Confronting the grey zone after severe brain injury

2019 ◽  
Vol 3 (6) ◽  
pp. 707-711 ◽  
Author(s):  
Andrew Peterson ◽  
Adrian M. Owen
Keyword(s):  
Brain Injury ◽  
Ethical Issues ◽  
Vegetative State ◽  
Clinical Care ◽  
Grey Zone ◽  
Severe Brain Injury ◽  
New Methods ◽  
Legal Decision Making ◽  
Technological Developments ◽  
The Brain

In recent years, rapid technological developments in the field of neuroimaging have provided several new methods for revealing thoughts, actions and intentions based solely on the pattern of activity that is observed in the brain. In specialized centres, these methods are now being employed routinely to assess residual cognition, detect consciousness and even communicate with some behaviorally non-responsive patients who clinically appear to be comatose or in a vegetative state. In this article, we consider some of the ethical issues raised by these developments and the profound implications they have for clinical care, diagnosis, prognosis and medical-legal decision-making after severe brain injury.

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