Fluorescent Substrates

6-Carboxytetramethyl rhodamine is a fluorescent dye that is used to measure the presence of cancer cells in urine samples. It binds to human protein and has been shown to be a useful probe for fluorescence measurements. 6-Carboxytetramethyl rhodamine has also been used as a fluorescence probe for the detection of proliferative diabetic retinopathy and michaelis-menten kinetics in cyclic peptides.

Fluorescent probe

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5-Carboxyfluorescein (5-FAM) is a self-quenching dye and one of the most common labelling agents for biomolecules, cell staining and localisation studies. The fluorescence is pH dependent, therefore it has been used to measure the internal pH of cells. As it is only membrane permeable to dead cells, it can be used as a staining agent to distinguish viable and non-viable cells. It is also used in PCR to label the 5’ end of oligonucleotides in the presence of a quencher at the 3’ end (dual labelled probe). During the amplification process, the dye is cleaved and the fluorescence increases proportionally with the amount of the specific sequence amplified during the PCR process. The development of the fluorescence signal is therefore specifically related to the amplification of the target sequence. The carboxylic acid can be activated to react with a primary amine.

Fluorescent reagent for labeling amines

Reactive fluorescent labeling reagent

Indocyanine green is a fluorescent dye with a variety of applications in medical imaging. It has been used to diagnose and evaluate the activity of cancer cells and brain functions. Indocyanine green has also been used as an angiographic agent for diagnosis of solid tumours. In addition, it can be used to evaluate liver function and the blood flow in the brain. The biological properties of indocyanine green have not yet been fully elucidated, but it is known that it binds to hepatocyte-like cells in culture and is found at high levels in human serum. This dye shows fluorescence when bound to DNA or RNA, which indicates its possible use as a molecular probe for detecting carcinoma cells.  Indocyanine green is a STT3B inhibitor and has been shown to prevent α-amanitin toxicity in mice (Wang et al, 2023). This means it could be a possible antidote to poisoning by Amanita, such as death cap mushroom Amanita phalloides, which is the cause of over 90% of fatal mushroom poisoning.

Brominated analog of carboxyfluorescein; singlet oxygen generator