Dyes and other colorants in microtechnique and biomedical research

Coloration, known in biological study as staining, enhances the visibility of living and dead objects under the microscope. This study reviews some of the colorants, chromogenic compounds and reactions exploited in staining, with special attention given to techniques that impart different colours to specific components of cells and tissues. A major mechanism, the first to be recognised, is the uptake of dye cations or anions by oppositely charged macromolecules. Modern investigations have shown that weaker electrical attractions are also important, especially in techniques that use more than one dye and methods for staining uncharged materials. The sizes and shapes of dye molecules or aggregates can also influence where they will lodge in a stained tissue. Reactive textile dyes have few uses in biological study, but there are histochemical methods that can covalently bind chromogens to DNA and to some macromolecular carbohydrates. Metal complexing dyes are universally used in microtechnique. Their mechanisms of action are not all fully understood, especially in the case of the routine haemalum and eosin (H & E) method that is routinely used for pathology. Staining a tiny quantity of a specific substance requires amplification to generate a visible signal. This is achieved with antibodies and catalysts. An immunoglobulin (antibody) molecule bound by a specific tissue component (antigen) can itself serve as antigen to more than one anti-immunoglubulin molecules that are labelled with an enzyme or with biotin. Enzyme labels catalyse the production of insoluble coloured materials. Biotin has a strong and specific affinity for either avidin or streptavidin, proteins with four biotin-binding sites that can be detected with biotinylated enzymes. Antibodies carried on colloidal gold particles are detected directly by electron microscopy or, for light microscopy, by a process known as physical development. The gold particles are catalytic nuclei for deposition of silver in chemical reactions that have also been used in black and white photography. 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The methods use of avidin that has been with a dye or an enzyme as but more avidin molecule has four biotin-binding one of has with a biotinylated the will be to with biotinylated amplification is in the method in a of avidin and biotinylated is the biotinylated of the molecules more than one and are to bind avidin molecules is not the enzyme for methods of is also used is a biotin-binding by is not a methods avidin is streptavidin, a more is are not the catalysts. amplification method used in and is the process known as physical an that is photography. The in an of of to of in a silver The catalyse the chemical of silver in the by the physical the silver is the and is achieved with a a silver and components as and complexing that the with deposition of first the sites of the and is the has and histochemical staining methods use of physical to catalytic sites in tissues. some methods for staining there is to that the catalytic silver are to be detected by electron microscopy in to proteins is also the of silver methods that nuclei than of silver can be by physical development. method and are with a that or are with a physical the sites of catalytic of in the tissue of have been that have been detected in and are for and methods can investigations of of gold particles are used to and proteins to of tissue for histochemical The molecules a on the of gold gold particles can be with different sizes in the of the of more than one with the electron microscope. The sizes are and gold particles are to be by light microscopy, they can be coloured in particles are catalytic for physical been used to in have been that the amplification of colloidal gold labels to visible staining There are a of reviews of colloidal gold in light and electron microscopy in the have been living and for some The have and colloidal and can also be taken A of and cells by mechanisms than and the staining is to staining is carried in the as a dye is the or a The staining to uptake of by cells that have been the but are all the dyes as are in in is used for of staining with the silver of the of the and the method is to silver staining by some for in can be used to and the The is not specific for The cells of the up the and can be by The of electrical in the cells is that the cells are for is in the and have used the dye to is one of dyes that are used to in the of has been staining the is not with the of specific living of the method are to The of uptake of the dye is requires the of in the but the or the is staining of can be achieved with and dyes the dye or and more be are for is in to the stained cells in the but the dye can be in by a is but are for of the that the stained with and The that have the of are an and has the and in the and in of to of the to in is to and compounds in of the but in dyes not to DNA and in living cells, is by the are staining is in nuclei or of the cells have the of the staining the and of living and dead cells or and are used in for staining of of and the of different of white and of are for of dyes by the the of compounds that can living cells and to is used for in of the of a to the staining of in and the in the and of cells to of the of the compounds that are used in to specific components of living cells are are also dyes or but are compounds for use in The also to that can and dead cells are used with cells, but they can also be to of living microscopy more of the than light microscopy the stained objects are light that on a is exploited also in techniques for and labelled cells in as or of dyes not living that are and with or dyes strong affinity for proteins or for the of the a in as the or The of a is by on as of the and the of the dyes not living cells, by anions on the of the dye cations are also the as in dead cells, and dyes can and to proteins or and are labels for dead cells, in and in stained by dyes by light of some dyes are uncharged and compounds have and the of living cells and are by The dye anions are and the compounds used in have a or that can with or and the dye to The is one with or with in of a as or in living cells can serve as visible of electrical as the of in the or in Staining methods for have been for they the that the are of The and of tissue the of to sites of in with that are to a of the of in the can be stained can be by the of in an a of the the that the is by the of an the are either by or by with a The is a than in for staining not on of silver and silver in the and The techniques of and and and The of silver by of is known to be to of The of silver by also is achieved by a of chemical that have been to or A major of is that a than the of to the of of to have more than is the of the silver techniques are to of has been as an to for some been with the of the silver methods have been in the study of to that proteins labelled or the enzyme a taken up the of and to the of the in the or The of by microscopy or by a histochemical that that of of the the enzyme taken the of and to the of the that to the the silver methods for and the method for in the methods of for the sites of of methods are on the of to insoluble coloured by of be as a of A major in the of can be in than of more by A is a that can to of are and will cells, they are also to for histochemical of the The first to be used as a This is to and is also taken up by and The of the in for in A that one of the of the in the and be detected the of as as in the This is not with is methods the of tiny of proteins and can be visible under the microscope. is the of a molecule is to bind the to cells of the be labelled with or detected is and up the of uptake and of labelled that the component not and to dyes as by by and that has affinity for DNA and to the nuclei of labelled the in the of the one and a of and to that in the of the that in sites the and the This been on the of by the of tissue are for some in a of and to a of silver a coloured of silver all the of a of the The of coloured has the study of and more a that covalently with one of the first for is and of a is to of a labelled with are for for of with as and than and compounds by and dyes and more than or bind to proteins and not the can be taken up by that are the in for and to in the the the can to especially in the for and are used than used as a as of chemical but shown to be by the and labels with that the dyes for by and The used are and dyes and living as either or in the for they are used for and of cells, in and in to the dyes are also the in they This of can be to dead and to tissue is a process and a of can The process is to by but the to is also with of the to cells to the dyes with are are with that are not by in are in a of sizes to and can be covalently with a of used by as a of The first use of for cells by and and used a of with or in of the of the also to the to be in the tissue by with The of labelled the molecules to for they are and the of and A of is for of is for of and The of is that will as a of to and visible to the by microscopy is to a to the of the can be by to the are and that the of is the the has been to be a more An is to the of to be to the production of a coloured This is carried by on the a of a and to the of the for an of an insoluble the sites of the The more an of is This known as is to labelled but is not for use with of as the used in the is a is detected with an or with a of avidin and biotinylated with biotinylated by and the has been by also labels are cells and not are for especially in the is and is or are also used for by and by uptake compounds have molecules than not of cells, but they can to by of are and are used as and and they are detected with avidin or streptavidin, labelled with either or a and have also been used for of in and This has on some biological staining for tiny to of specific and the use of dyes and for and in living cells and There are of biological staining and that not be in an of The of coloured and compounds for in in is is a the of where has for has four and more than and and to and The is also an with a few in that

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