Multi-well plate

The model immunoassay is the enzyme-linked immunosorbent assay (ELISA) in which a non-specific capture antibody is bound to a surface, such as a multi-well plate or small tube [13]. In the basic form of ELISA, a second antibody tagged with an enzyme interacts specifically with the analyte. The enzyme assay produces a colored product that is read with a spectrophotometer. There are many variations on the basic immunoassay format that serve to increase sensitivity, specificity, linear range, and speed. Many commercial instruments have been developed to take advantage of various technologies for reporter molecules. The immunoassay may be coupled to an electronic sensor and transducer, such as a surface acoustical wave (SAW) sensor. Electrochemiluminescence (ECL) is a method in which the detector antibody is tagged with a ruthenium-containing chelate [13-15]. When the tag is... 

The choice of cell culture method for cytotoxicity studies for a certain substance depends on the target cells and assay duration. Organ, spheroids, suspension cells, flask surface, and multi-well plate monolayers, as well as agar surface cultures can be employed. 

Clear, sterile, tissue culture treated multi-well plates (6-well, 12-well, or 24-well, etc.)... 

Cell plating. Twenty-four hours prior to transfection, seed cells into a clear tissue culture multi-well plate at 150 cclls/pL according to Table 1. 

Hahn, M.E., B.L. Woodward, J.J. Stegeman and S.W. Kennedy. Rapid assessment of induced cytochrome P4501A (CYP1A) protein and catalytic activity in fish hepatoma cells grown in multi-well plates response to TCDD, TCDF and planar PCBs. Environ. Toxicol. Chem. 15 582-591, 1996. 

For routine assays requiring analysis of large numbers of samples, micro-titre or multi-well plate readers have been developed. These consist of single-beam instruments in which the samples are contained in standard 96-well (8x12) plates, each well containing a separate sample. This allows large numbers of samples to be measured rapidly, often automatically under robotic control. The multi-well/micro-titre plates themselves are often coated with specific reagents so that particular analyses may be done rapidly and routinely in situ. 

Fig. 3.6 Schematic diagram of a multi-spot plate assay for four human cytokines. Each spot within each well of the multi-well plate contains capture antibody specific for one cytokine. (Inset) Images of the ECL emitted from assays and protein immobilized on carbon electrode surface (Reprinted with permission from Ref. [10]. Copyright 2008 American Chemical Society)...

Ressler, J., H. Grothe, E. Motiescu, and B. Wolf (2004). New concepts for chip-supported multi-well-plates realization of a 24-well-plate with integrated impedance-sensors for functional cellular screening applications and automated microscope aided cell-based assays. In Engineering in Medicine and Biology Society, 2004. IEMBS 04. 26th Annual International Conference of the IEEE, Volume 1, pp. 2074-2077. IEEE. 

A standard multi-well plate reader (we use BIO-RAD Model 680 Microplate Reader) with absorbance measurement capability at 490 nm. 

Cell proliferation is quantified by using the Aqueous Non-Radioactive Cell Proliferation Assay according to the manufacturer s instructions and absorbance of the wells is measured using a standard multi-well plate reader. 

One strategy makes use of arrays of molecules that are synthesized, e.g., on a chip or a multiwell reactor. Then the biological entity for which a ligand is to be selected (e.g., a membrane receptor, an enzyme, a structural protein...) is bound to the chip or the multi-well plate. The sites to which strong binding occurs are identiried, and, consequently, as the array has been synthesized in a controlled order, the best ligand is identified. 

Stamped copies of agar plates Duplicates of multi welled plates... 

Cells can be induced to fuse if two cell populations are brought close together at a high cell concentration (10 6 to 10 7 cells per well of a 96 multi-well plate) in the presence of viruses or by chemical agents (called fuso-gens ). The process involves a destabilization of adjacent cell membranes which eventually fuse to form a hybrid cell. Initially, two distinct nuclei are present in the fused cell (a heterokaryon). Eventually the nuclei fuse to produce a stable hybrid cell. 

ELISA is the most commonly used assay for antibodies and is adapted to multi-well plates for analyzing multiple samples. RIA is more sensitive but is more time consuming and expensive. Affinity chromatography is ideal if a high-performance liquid chromatography (HPLC) system is available and the hybridomas are growing in a serum-ffee medium. The basis of the three types of assay are described here ... 

Diagnostic applications may require standardized samples, e.g. multi-well plates with larger numbers of seeded cells. Therefore, a fluorescence reader with multiple total internal reflection (TIR) was developed to examine the plasma membranes of 96 cell collectives simultaneously [52], as shown in Figure 9. Fluorescence spectroscopy and microscopy are indispensable prerequisites for those studies. 

Hepatocytes were isolated from rats by in situ perfusion of the liver with collagenase solution by the method of Tanaka et ah 14), Isolated hepatocytes were suspended at a concentration of 5 x 10 cells/ml in William s medium E with 1 nM insulin, 1 nM dexamethasone, 100 mg/1 kanamycin, 10 klU/ml aprotinin and 5% fetal bovine serum. The cells were seeded on plastic multi-well plates or dishes (Becton Dickinson Co., Ltd., Franklin Lakes, NJ) pre-coated with collagen type I, then cultured under an atmosphere of 95% air-5% CO2 at 37°C for 2 h or 20h. Hepatocytes were treated with various concentrations of Trp-P-1 in dimethyl sulfoxide (DMSO) for various times as indicated in each figure. Parallel dishes were treated with vehicle alone to obtain control sanq>les (maximum concentration of DMSO in the medium was 0.1%, v/v). 

Another test performed was the observation of the biodegradation test over a period of 30 days. Constructs (n=5) were placed in a multi-well plate and incubated at 37° C with 5% purified CO2. The growth medium was changed every 2 days (Eigure 5). 

Anovel screening method using multi-well plates has been reported [4]. Microbial cultures of about 300microbes, demonstrating utility in reducing model ketones, were arrayed in multi-well plates, stored until use at -80 °C, and used to rapidly... 

Screening of biocatalysts (a) from culturable microorganisms using multi-well plates (b) using metagenomes of complex... 

Remove the culture medium from a petri dish containing the coverslip with attached cells or the slices on the Millicell-CM insert and add 1 mL of 4 % PFA in PB. Fix for 1 h and wash 3x5 min in PBS. From this stage on cultures can be processed in multi-well plates by transferring them from a well to another with forceps. Isolated cells attached to coverslips can be directly handled as indicated, whereas organotypic cultures need to be separated from the Millicell-CM insert by carefully cutting the membrane around tissue slices and leaving enough membrane to allow manipulation. 

When culturing in 96-well multi-well plates, the seeding density of T. vaginalis is 5x10 cells per well, added in 100 pL MDM from a suspension of 5 x 10 trophozoites/mL medium. 

Briefly spin down the compound source plates at 300 for 2 min in a tabletop centrifuge equipped with multi-well plate adaptor. 

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