Electroporation
Electroporation, also known as electropermeabilization, is a microbiological and biotechnological technique in which an electric field is applied to cells to briefly increase the permeability of the cell membrane. The application of a high-voltage electric field induces a temporary destabilization of the lipid bilayer, resulting in the formation of nanoscale pores that permit the entry or exit of macromolecules. This method is widely employed to introduce molecules—including small molecules, DNA, RNA, and proteins—into cells. Electroporation can be performed on cells in suspension using electroporation cuvettes, or directly on adherent cells ''in situ'' within their culture vessels. In microbiology, electroporation is frequently utilized for the transformation of bacteria or yeast cells, often with plasmid DNA. It is also used in the transfection of plant protoplasts and mammalian cells. Notably, electroporation plays a critical role in the '' ex vivo'' manipulation ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |
|
Electroporation Cuvettes
Electroporation, also known as electropermeabilization, is a Microbiology, microbiological and Biotechnology, biotechnological technique in which an electric field is applied to Cell (biology), cells to briefly increase the Permeability (porous media), permeability of the cell membrane. The application of a high-voltage electric field induces a temporary destabilization of the lipid bilayer, resulting in the formation of nanoscale pores that permit the entry or exit of Macromolecule, macromolecules. This method is widely employed to introduce molecules—including small molecules, DNA, RNA, and proteins—into cells. Electroporation can be performed on cells in suspension using electroporation Cuvette, cuvettes, or directly on adherent cells ''in situ'' within their culture vessels. In microbiology, electroporation is frequently utilized for the Genetic transformation, transformation of bacteria or yeast cells, often with plasmid DNA. It is also used in the transfection of plant ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |
|
Lipid Bilayer
The lipid bilayer (or phospholipid bilayer) is a thin polar membrane made of two layers of lipid molecules. These membranes form a continuous barrier around all cell (biology), cells. The cell membranes of almost all organisms and many viruses are made of a lipid bilayer, as are the nuclear envelope, nuclear membrane surrounding the cell nucleus, and biological membrane, membranes of the membrane-bound organelles in the cell. The lipid bilayer is the barrier that keeps ions, proteins and other molecules where they are needed and prevents them from diffusing into areas where they should not be. Lipid bilayers are ideally suited to this role, even though they are only a few nanometers in width, because they are impermeable to most water-soluble (hydrophilic) molecules. Bilayers are particularly impermeable to ions, which allows cells to regulate salt concentrations and pH by transporting ions across their membranes using proteins called Ion transporter, ion pumps. Biological bilaye ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |
|
Tissue Nanotransfection
Tissue nanotransfection (TNT) is an electroporation-based technique capable of gene and drug cargo delivery or transfection at the nanoscale. Furthermore, TNT is a scaffold-less tissue engineering (TE) technique that can be considered cell-only or tissue inducing depending on cellular or tissue level applications. The transfection method makes use of nanochannels to deliver cargo to tissues topically. History Cargo delivery methods rely on carriers, for example nanoparticles, viral vectors, or physical approaches such as gene guns, microinjection, or electroporation. The various methods can be limited by size constraints or their ability to efficiently deliver cargo without damaging tissue. Electroporation is a physical method which harnesses an electric field to open pores in the normally semi-permeable cell membrane through which cargo can enter. In this process, the charges can be used to drive cargo in a specific direction. Bulk electroporation (BEP) is the most conventio ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |
|
Genetic Transformation
In molecular biology and genetics, transformation is the genetic alteration of a cell resulting from the direct uptake and incorporation of exogenous genetic material from its surroundings through the cell membrane(s). For transformation to take place, the recipient bacterium must be in a state of competence, which might occur in nature as a time-limited response to environmental conditions such as starvation and cell density, and may also be induced in a laboratory. Transformation is one of three processes that lead to horizontal gene transfer, in which exogenous genetic material passes from one bacterium to another, the other two being conjugation (transfer of genetic material between two bacterial cells in direct contact) and transduction (injection of foreign DNA by a bacteriophage virus into the host bacterium). In transformation, the genetic material passes through the intervening medium, and uptake is completely dependent on the recipient bacterium. As of 2014 about 8 ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |
|
Calcium Chloride Transformation
Calcium chloride (CaCl2) transformation is a laboratory technique in prokaryotic (bacterial) cell biology. The addition of calcium chloride to a cell suspension promotes the binding of plasmid DNA to lipopolysaccharides (LPS). Positively charged calcium ions attract both the negatively charged DNA backbone and the negatively charged groups in the LPS inner core. The plasmid DNA can then pass into the cell upon heat shock, where chilled cells (+4 degrees Celsius) are heated to a higher temperature (+42 degrees Celsius) for a short time. History of bacterial transformation Frederick Griffith published the first report of bacteria's potential for transformation in 1928. Griffith observed that mice did not succumb to the "rough" type of ''pneumococcus'' (''Streptococcus pneumoniae''), referred to as nonvirulent, but did succumb to the "smooth" strain, which is referred to as virulent. The smooth strain's virulence could be suppressed with heat-killing. However, when the nonvirulent ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |
|
Transfection
Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. It may also refer to other methods and cell types, although other terms are often preferred: " transformation" is typically used to describe non-viral DNA transfer in bacteria and non-animal eukaryotic cells, including plant cells. In animal cells, transfection is the preferred term, as the term "transformation" is also used to refer to a cell's progression to a cancerous state (carcinogenesis). Transduction is often used to describe virus-mediated gene transfer into prokaryotic cells. The word ''transfection'' is a portmanteau of the prefix ''trans-'' and the word "infection." Genetic material (such as supercoiled plasmid DNA or siRNA constructs), may be transfected. Transfection of animal cells typically involves opening transient pores or "holes" in the cell membrane to allow the uptake of material. Transfection can be carried out using calcium phosphate (i.e. ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |
|
Peptide
Peptides are short chains of amino acids linked by peptide bonds. A polypeptide is a longer, continuous, unbranched peptide chain. Polypeptides that have a molecular mass of 10,000 Da or more are called proteins. Chains of fewer than twenty amino acids are called oligopeptides, and include dipeptides, tripeptides, and tetrapeptides. Peptides fall under the broad chemical classes of biological polymers and oligomers, alongside nucleic acids, oligosaccharides, polysaccharides, and others. Proteins consist of one or more polypeptides arranged in a biologically functional way, often bound to ligands such as coenzymes and cofactors, to another protein or other macromolecule such as DNA or RNA, or to complex macromolecular assemblies. Amino acids that have been incorporated into peptides are termed residues. A water molecule is released during formation of each amide bond.. All peptides except cyclic peptides have an N-terminal (amine group) and C-terminal (carboxyl g ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |
|
Miniaturization
Miniaturization ( Br.Eng.: ''miniaturisation'') is the trend to manufacture ever-smaller mechanical, optical, and electronic products and devices. Examples include miniaturization of mobile phones, computers and vehicle engine downsizing. In electronics, the exponential scaling and miniaturization of silicon MOSFETs (MOS transistors) leads to the number of transistors on an integrated circuit chip doubling every two years, an observation known as Moore's law. This leads to MOS integrated circuits such as microprocessors and memory chips being built with increasing transistor density, faster performance, and lower power consumption, enabling the miniaturization of electronic devices. Electronic circuits The history of miniaturization is associated with the history of information technology based on the succession of switching devices, each smaller, faster, and cheaper than its predecessor. During the period referred to as the Second Industrial Revolution (), miniaturizati ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |
|
Viability Assay
A viability assay is an assay that is created to determine the ability of organs, cells or tissues to maintain or recover a state of survival. Viability can be distinguished from the all-or-nothing states of life and death by the use of a quantifiable index that ranges between the integers of 0 and 1 or, if more easily understood, the range of 0% and 100%. Viability can be observed through the physical properties of cells, tissues, and organs. Some of these include mechanical activity, motility, such as with spermatozoa and granulocytes, the contraction of muscle tissue or cells, mitotic activity in cellular functions, and more. Viability assays provide a more precise basis for measurement of an organism's level of vitality. Viability assays can lead to more findings than the difference of living versus nonliving. These techniques can be used to assess the success of cell culture techniques, cryopreservation techniques, the toxicity of substances, or the effectiveness of substanc ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |
|
Gene Gun
In genetic engineering, a gene gun or biolistic particle delivery system is a device used to deliver exogenous DNA (transgenes), RNA, or protein to cells. By coating particles of a heavy metal with a gene of interest and firing these micro-projectiles into cells using mechanical force, an integration of desired genetic information can be introduced into desired cells. The technique involved with such micro-projectile delivery of DNA is often referred to as biolistics, short for "biological ballistics". This device is able to transform almost any type of cell and is not limited to the transformation of the nucleus; it can also transform organelles, including plastids and mitochondria. Gene gun design The gene gun was originally a Crosman air pistol modified to fire dense tungsten particles. It was invented by John C Sanford, Ed Wolf, and Nelson Allen at Cornell University along with Ted Klein of DuPont between 1983 and 1986. The original target was onions (chosen for their ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |
|
Microinjection
Microinjection is the use of a glass micropipette to inject a liquid substance at a microscopic or borderline macroscopic level. The target is often a living cell but may also include intercellular space. Microinjection is a simple mechanical process usually involving an inverted microscope with a Optical power, magnification power of around 200x (though sometimes it is performed using a dissecting stereo microscope at 40–50x or a traditional Optical microscope#Compound microscope, compound upright microscope at similar power to an inverted model). For processes such as cellular or pronucleus, pronuclear injection the target cell is positioned under the microscope and two micromanipulation, micromanipulators—one holding the pipette and one holding a microcapillary needle usually between 0.5 and 5 μm in diameter (larger if injecting stem cells into an embryo)—are used to penetrate the cell membrane and/or the nuclear envelope. In this way the process can be used to int ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |
|
In Vivo
Studies that are ''in vivo'' (Latin for "within the living"; often not italicized in English) are those in which the effects of various biological entities are tested on whole, living organisms or cells, usually animals, including humans, and plants, as opposed to a tissue extract or dead organism. Examples of investigations ''in vivo'' include: the pathogenesis of disease by comparing the effects of bacterial infection with the effects of purified bacterial toxins; the development of non-antibiotics, antiviral drugs, and new drugs generally; and new surgical procedures. Consequently, animal testing and clinical trials are major elements of ''in vivo'' research. ''In vivo'' testing is often employed over ''in vitro'' because it is better suited for observing the overall effects of an experiment on a living subject. In drug discovery, for example, verification of efficacy ''in vivo'' is crucial, because ''in vitro'' assays can sometimes yield misleading results with drug c ... [...More Info...]       [...Related Items...]     OR:     [Wikipedia]   [Google]   [Baidu]   |