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A nitrocellulose slide (or nitrocellulose film slide) is a glass
microscope slide A microscope slide is a thin flat piece of glass, typically 75 by 26 mm (3 by 1 inches) and about 1 mm thick, used to hold objects for examination under a microscope. Typically the object is mounted (secured) on the slide, and then b ...
that is coated with
nitrocellulose Nitrocellulose (also known as cellulose nitrate, flash paper, flash cotton, guncotton, pyroxylin and flash string, depending on form) is a highly flammable compound formed by nitrating cellulose through exposure to a mixture of nitric acid and ...
that is used to bind biological material, often
protein Proteins are large biomolecules and macromolecules that comprise one or more long chains of amino acid residues. Proteins perform a vast array of functions within organisms, including catalysing metabolic reactions, DNA replication, res ...
, for colorimetric and fluorescence detection assays. For this purpose, a nitrocellulose slide is generally considered to be superior to glass, because it binds a great deal more protein, and protects the
tertiary structure Protein tertiary structure is the three dimensional shape of a protein. The tertiary structure will have a single polypeptide chain "backbone" with one or more protein secondary structures, the protein domains. Amino acid side chains may i ...
of the protein (and other biological material, i.e.: cells). Typically, nitrocellulose slides have a thin, opaque film of nitrocellulose on a standard 25mm × 75 mm glass
microscope slide A microscope slide is a thin flat piece of glass, typically 75 by 26 mm (3 by 1 inches) and about 1 mm thick, used to hold objects for examination under a microscope. Typically the object is mounted (secured) on the slide, and then b ...
. The film is extremely sensitive to contact, and to foreign material; contact causes deformation and deposition of material, especially liquids. A nitrocellulose slide is different from a nitrocellulose membrane, which usually filters protein from solution (i.e.: physician's office pregnancy tests), but that it serves a similar goal: to detect the presence and/or concentration level of certain biological material.


Microarrays

Nitrocellulose slides are used mainly in
proteomics Proteomics is the large-scale study of proteins. Proteins are vital parts of living organisms, with many functions such as the formation of structural fibers of muscle tissue, enzymatic digestion of food, or synthesis and replication of DNA. I ...
to do protein microarrays with automated systems that print the slides and record results. Microarrays of cell analytes, arrays of
cell lysate Lysis ( ) is the breaking down of the membrane of a cell, often by viral, enzymic, or osmotic (that is, "lytic" ) mechanisms that compromise its integrity. A fluid containing the contents of lysed cells is called a ''lysate''. In molecular bi ...
,
antibody An antibody (Ab), also known as an immunoglobulin (Ig), is a large, Y-shaped protein used by the immune system to identify and neutralize foreign objects such as pathogenic bacteria and viruses. The antibody recognizes a unique molecule of t ...
microarrays, tissue printing, immunoarrays, etc. are also possible with the slide.


Nitrocellulose fluorescence

Due to their high surface roughness, conventional white nitrocellulose films scatter and reflect large amounts of excitation and emission light during the fluorescence detection in the microarray scanner. In addition, nitrocellulose exhibits a natural autofluorescence at the detection wavelengths commonly used. Both these factors lead to a high background fluorescent signal from these membrane slides. To overcome this problem, a new process has been developed to generate black membranes that absorb the scattered light, significantly reducing the background auto-fluorescence and thus offering a very low and homogenous auto-fluorescence to achieve a significantly improved dynamic range. These slides are commercially available through
Schott AG Schott AG is a German multinational glass company specializing in the manufacture of glass and glass-ceramics. Headquartered in Mainz, Germany, it is owned by the Carl Zeiss Foundation. The company's founder and namesake, Otto Schott, is cre ...
. Nevertheless, conventional white nitrocellulose films continue to be the dominant surface for many protein microarray applications because the claims above have not proved relevant to end user requirements. Regardless, nitrocellulose slide manufacturers like Grace Bio-Labs continue to develop new nitrocellulose surfaces to further optimize their use in protein microarrays. A method for protein quantitation on nitrocellulose coated glass slides uses near-IR fluorescent detection with
quantum dot Quantum dots (QDs) are semiconductor particles a few nanometres in size, having optical and electronic properties that differ from those of larger particles as a result of quantum mechanics. They are a central topic in nanotechnology. When the ...
s. Traditional porous nitrocellulose signal to noise is limited by auto-fluorescence of the nitrocellulose at the respective required wavelengths of excitation and emission for standard organic fluorescent detection probes. Near IR detection probes are excited and read at emission wavelengths outside the range of nitrocellulose fluorescence.Marie‐Laure Schneider, Adriana Marquez‐ Lagraulet, Richard Pasquesi, Michael Shultz - (2014)
"Infrared detection decreases nitrocellulose auto-fluorescence and improves RPPA assays signal‐to‐noise ratio over visible wavelength detection", Innopsys Inc. Chicago, Illinois


References

{{Reflist Microscopy Biochemistry methods