Background Fluorescence Interference Research Articles

A near-infrared fluorescent probe for lipid hydroperoxides in living cells

An NIR (near-infrared) fluorescent probe TCP (tricarbocyanine diphenylphosphine) including a non-conjugated 'pre-tricarbocyanine' was designed and synthesized for visualizing lipid hydroperoxides (ROOH) in living cells. The excitation and emission spectra of tricarbocyanine in the NIR region can effectively avoid background fluorescence interference in biological systems. The probe exhibited a rapid fluorescence response to ROOH and high selectivity for ROOH over other ROS (reactive oxygen species) and some biological compounds, and the limit of detection was 38 pM. In addition, the probe was stable, and less cytotoxic, which indicated that it has potential application in detecting lipid hydroperoxides in living biological systems.

A Phosphinate‐Based Red Fluorescent Probe for Imaging the Superoxide Radical Anion Generated by RAW264.7 Macrophages

4',9'-Bis(diphenylphosphinyl)naphthofluorescein (PNF-1) has been designed and synthesized as a highly selective, sensitive, cell-permeable, red fluorescent probe for detecting O(2) (.-) in biological systems. The design strategy for the probe is based on the nucleophilic mechanism of O(2) (.-) to mediate deprotection of the probe to naphthofluorescein, the emission spectrum of which is just in the spectral region of low background fluorescence interference in biological systems. Upon treatment with O(2) (.-), the probe exhibits a strong fluorescence response and high selectivity for O(2) (.-), rather than other reactive oxygen species or biological compounds. A linear calibration curve for PNF-1 showed a detection limit of 0.1 nM O(2) (.-). This new type of fluorescent probe allows nanomolar changes in O(2) (.-) concentrations in living cells to be detected by confocal microscopy.

Laser Raman spectrum of calcified human aorta.

Raman spectra of highly calcified areas of human aorta were obtained using long wavelength excitation at 740 nm to minimize background fluorescence interference. Raman spectra resembling that of hydroxyapatite were obtained for segments of highly calcified aorta that were thawed frozen samples, from which the surface layer had been removed, or were dried. These are the first reported spectra obtained in air or in normal saline solution showing the hydroxyapatite peak in calcified plaque for samples which were untreated except for freezing, which is encouraging for the application of Raman spectroscopy as a method of detection of plaque during laser angioplasty. Spectra obtained from calcified aorta using excitation at 514.5 nm did not show evidence of the hydroxyapatite band because of the severe interference by background fluorescence generated at this wavelength.