Accelerated transgene expression of pDNA/polysaccharide complexes by solid-phase reverse transfection and analysis of the cell transfection mechanism

2022 ◽  
Author(s):  
Takahiro Arai ◽  
Yu Aiki ◽  
Toshinori Sato
Keyword(s):  
Transgene Expression ◽  
Solid Phase ◽  
Cell Transfection ◽  
Reverse Transfection

scholarly journals Spatial Control of Cell Transfection Using Soluble or Solid-Phase Redox Agents and a Redox-Active Ferrocenyl Lipid

2013 ◽  
Vol 5 (17) ◽  
pp. 8283-8288 ◽  
Author(s):  
Burcu S. Aytar ◽  
John P. E. Muller ◽  
Yukishige Kondo ◽  
Nicholas L. Abbott ◽  
David M. Lynn
Keyword(s):  
Solid Phase ◽  
Cell Transfection ◽  
Spatial Control ◽  
Redox Active ◽  
Redox Agents

scholarly journals Work Flow for Multiplexing siRNA Assays by Solid-Phase Reverse Transfection in Multiwell Plates

2008 ◽  
Vol 13 (7) ◽  
pp. 575-580 ◽  
Author(s):  
Holger Erfle ◽  
Beate Neumann ◽  
Phill Rogers ◽  
Jutta Bulkescher ◽  
Jan Ellenberg ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Solid Phase ◽  
Transfection Efficiency ◽  
Rt Pcr ◽  
Flexible Tool ◽  
Cellular Assays ◽  
Long Term Storage ◽  
Cell Microarrays ◽  
Reverse Transfection ◽  
Multiwell Plates

Solid-phase reverse transfection on cell microarrays is a high-throughput method for the parallel transfection of mammalian cells. However, the cells transfected in this way have been restricted so far to microscopy-based analyses. Analysis methods such as reverse transcriptase—polymerase chain reaction (RT-PCR) and access to higher cell numbers for statistical reasons in microscopy-based assays are not possible with solid-phase reverse transfection on cell microarrays. We have developed a quick and reliable protocol for automated solid-phase reverse transfection of human cells with siRNAs in multiwell plates complementing solid-phase reverse transfection on cell microarrays. The method retains all advantages of solid-phase reverse transfection such as long-term storage capacity after fabrication, reduced cytotoxicity, and reduced cost per screen compared with liquid-phase transfection in multiwell plates. The protocol has been tested for the RNAi-mediated knockdown of several genes in different cell lines including U20S, RPE1, A549, and HeLa cells. We show that even 3 months after production of the “ready to transfect” multiwell plates, there is no reduction in their transfection efficiency as assessed by RT-PCR and nuclear phenotyping by fluorescence microscopy. We conclude that solid-phase reverse transfection in multiwell plates is a cost-efficient and flexible tool for multiplexing cellular assays. ( Journal of Biomolecular Screening. 2008:575-580)

scholarly journals Solid-phase reverse transfection for intracellular delivery of functionally active proteins

2017 ◽  
Vol 27 (10) ◽  
pp. 1752-1758 ◽  
Author(s):  
Ruben Bulkescher ◽  
Vytaute Starkuviene ◽  
Holger Erfle
Keyword(s):  
Solid Phase ◽  
Intracellular Delivery ◽  
Reverse Transfection

Solid-phase immunoelectron microscopy for rapid diagnosis of enteroviruses

1984 ◽  
Vol 42 ◽  
pp. 226-227 ◽  
Author(s):  
K. Pegg-Feige ◽  
F. W. Doane
Keyword(s):  
Electron Microscopy ◽  
Cell Culture ◽  
Immunoelectron Microscopy ◽  
Detection Method ◽  
Solid Phase ◽  
Protein A ◽  
Plant Viruses ◽  
Rapid Diagnosis ◽  
Virus Diagnosis ◽  
Antiviral Antibody

Immunoelectron microscopy (IEM) applied to rapid virus diagnosis offers a more sensitive detection method than direct electron microscopy (DEM), and can also be used to serotype viruses. One of several IEM techniques is that introduced by Derrick in 1972, in which antiviral antibody is attached to the support film of an EM specimen grid. Originally developed for plant viruses, it has recently been applied to several animal viruses, especially rotaviruses. We have investigated the use of this solid phase IEM technique (SPIEM) in detecting and identifying enteroviruses (in the form of crude cell culture isolates), and have compared it with a modified “SPIEM-SPA” method in which grids are coated with protein A from Staphylococcus aureus prior to exposure to antiserum.

Application of solid-phase immune electron microscopy to study physical and stability characteristics of enterically transmitted non-a, non-b hepatitis virus

1988 ◽  
Vol 46 ◽  
pp. 80-81
Author(s):  
Charles D. Humphrey ◽  
E. H. Cook ◽  
Karen A. McCaustland ◽  
Daniel W. Bradley
Keyword(s):  
Electron Microscopy ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Etiologic Agent ◽  
World Health ◽  
Health Concern ◽  
Morphological Identification ◽  
Immune Electron Microscopy

Enterically transmitted non-A, non-B hepatitis (ET-NANBH) is a type of hepatitis which is increasingly becoming a significant world health concern. As with hepatitis A virus (HAV), spread is by the fecal-oral mode of transmission. Until recently, the etiologic agent had not been isolated and identified. We have succeeded in the isolation and preliminary characterization of this virus and demonstrating that this agent can cause hepatic disease and seroconversion in experimental primates. Our characterization of this virus was facilitated by immune (IEM) and solid phase immune electron microscopic (SPIEM) methodologies.Many immune electron microscopy methodologies have been used for morphological identification and characterization of viruses. We have previously reported a highly effective solid phase immune electron microscopy procedure which facilitated identification of hepatitis A virus (HAV) in crude cell culture extracts. More recently we have reported utilization of the method for identification of an etiologic agent responsible for (ET-NANBH).

Identification of hepatitis a virus in cell cultures by solid phase immune electron microscopy

1986 ◽  
Vol 44 ◽  
pp. 238-239
Author(s):  
C.D. Humphrey ◽  
T.L. Cromeans ◽  
E.H. Cook ◽  
D.W. Bradley
Keyword(s):  
Electron Microscopy ◽  
Liquid Phase ◽  
Cell Cultures ◽  
Rapid Detection ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Immune Electron Microscopy ◽  
Detection And Identification

There is a variety of methods available for the rapid detection and identification of viruses by electron microscopy as described in several reviews. The predominant techniques are classified as direct electron microscopy (DEM), immune electron microscopy (IEM), liquid phase immune electron microscopy (LPIEM) and solid phase immune electron microscopy (SPIEM). Each technique has inherent strengths and weaknesses. However, in recent years, the most progress for identifying viruses has been realized by the utilization of SPIEM.

Dynamic studies of silicide-mediated crystallization of amorphous silicon

1992 ◽  
Vol 50 (2) ◽  
pp. 1352-1353
Author(s):  
C. Hayzelden ◽  
J. L. Batstone
Keyword(s):  
Ion Implantation ◽  
Solid Phase ◽  
Metallic Phase ◽  
Single Crystal Substrate ◽  
Free Electrons ◽  
Melt Temperature ◽  
Transmission Electron ◽  
Crystal Substrate ◽  
High Resolution Tem

Epitaxial reordering of amorphous Si(a-Si) on an underlying single-crystal substrate occurs well below the melt temperature by the process of solid phase epitaxial growth (SPEG). Growth of crystalline Si(c-Si) is known to be enhanced by the presence of small amounts of a metallic phase, presumably due to an interaction of the free electrons of the metal with the covalent Si bonds near the growing interface. Ion implantation of Ni was shown to lower the crystallization temperature of an a-Si thin film by approximately 200°C. Using in situ transmission electron microscopy (TEM), precipitates of NiSi2 formed within the a-Si film during annealing, were observed to migrate, leaving a trail of epitaxial c-Si. High resolution TEM revealed an epitaxial NiSi2/Si(l11) interface which was Type A. We discuss here the enhanced nucleation of c-Si and subsequent silicide-mediated SPEG of Ni-implanted a-Si.Thin films of a-Si, 950 Å thick, were deposited onto Si(100) wafers capped with 1000Å of a-SiO2. Ion implantation produced sharply peaked Ni concentrations of 4×l020 and 2×l021 ions cm−3, in the center of the films.

TEM characterization of Au/Polysilicon interactions

1990 ◽  
Vol 48 (4) ◽  
pp. 650-651
Author(s):  
N. David Theodore ◽  
Leslie H. Allen ◽  
C. Barry Carter ◽  
James W. Mayer
Keyword(s):  
Solid Phase ◽  
Single Crystal Silicon ◽  
Chemical Vapor ◽  
Electrical Contacts ◽  
Silicon Substrates ◽  
Crystal Silicon ◽  
Transmission Electron ◽  
Polysilicon Films ◽  
The Stability

Metal/polysilicon investigations contribute to an understanding of issues relevant to the stability of electrical contacts in semiconductor devices. These investigations also contribute to an understanding of Si lateral solid-phase epitactic growth. Metals such as Au, Al and Ag form eutectics with Si. reactions in these metal/polysilicon systems lead to the formation of large-grain silicon. Of these systems, the Al/polysilicon system has been most extensively studied. In this study, the behavior upon thermal annealing of Au/polysilicon bilayers is investigated using cross-section transmission electron microscopy (XTEM). The unique feature of this system is that silicon grain-growth occurs at particularly low temperatures ∽300°C).Gold/polysilicon bilayers were fabricated on thermally oxidized single-crystal silicon substrates. Lowpressure chemical vapor deposition (LPCVD) at 620°C was used to obtain 100 to 400 nm polysilicon films. The surface of the polysilicon was cleaned with a buffered hydrofluoric acid solution. Gold was then thermally evaporated onto the samples.

New results on the solid-phase recrystallisation of ZnSe

1998 ◽  
Vol 184-185 (1-2) ◽  
pp. 1021-1025
Author(s):  
G Geoffroy
Keyword(s):  
Solid Phase
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