Overview of Technology
Determine instrument polarization bias in detection of the polarized scattered, transmitted, and emitted lights.
A set of fluorescent mixture samples for determination and validation of instrument polarization bias in detection of the polarized scattered, transmitted and emitted lights. Many optical measurements rely on reliable data acquisition conducted with photon excitation and detection. These include quantification of the depolarization and anisotropy of the sample stokes-shifted fluorescence, on resonance fluorescence anisotropy and material photon scattering. However, the light throughput of many optical elements includes the lens, mirrors, polarizers and polarization dependent referred to as the instrument polarized bias.
Many optical measurements rely on reliable data acquisition conducted with polarized photon excitation and detection. These include the quantification of the depolarization and anisotropy of the sample stokes-shifted fluorescence, on resonance fluorescence anisotropy and material photon scattering. However, the light throughput of many optical elements includes the lens, mirrors, polarizers and polarization dependent referred to as the instrument polarized bias.
One must reliably compensate the instrument polarization bias in order to obtain the optical anisotropy for the sample. This correction is commonly achieved through a wavelength-dependent G-factor used in the fluorescence spectra for solution that contain the fluorophore of interest. With such approach, only a single wavelength or narrow wavelength range of the G-factors can be determined, and the reliability of the G-factor can’t be independently validated. Any problem with this G-factor determination is that the possible photon bleaching or other photon chemical reaction during the g-factor quantification step can compromise the reliability of experimental results of the subsequent measurement conducted for the same sample of interest.
This invention pertains to a set of fluorescent samples for determination and validation of instrument polarization bias in detection of the polarized, scattered, transmitted, and emitted lights. Additionally, there is a new type of optical instrument for single and multi-wavelength studying matter/light interactions.
This invention pertains to a set of fluorescence samples that can be used for determination and independent validation of the G-factors with a broad wavelength region. These samples are referred to hereafter as G-factor samples or G-factor calibration and validation samples. The G-factor sample with photons at different wavelengths, the g-factor sample emit stokes-shifted fluorescence spanning a wider wavelength region that is impossible by individual fluorophores. Therefore, with one specifically designed G-factor sample, the g-factor can be quantified with a broader wavelength region.
The design of a fix-wavelength photon scattering, on-resonance emission and stokes-shifted fluorescence detection proposes a design that aims to achieve stokes-shifted fluorescence, by setting the detection wavelength longer than the excitation wavelength.