Rapid, massively parallel single-cell drug response measurements via live cell interferometry
A technology of interferometry and living cells, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., and can solve the problems of lack of understanding of the size of individual cells
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[0052] Other exemplary embodiments of the invention can include, for example, systems and methods for quantifying cell masses and / or observing how one or more cell masses change in response to environmental stimuli. Some embodiments include a method comprising the steps of: placing one or more cells in an observation chamber of an interferometric microscope capable of producing and measuring a fractional phase shift between a test beam and a reference beam; exposing the cells to of the test beam; measuring the fractional phase shift between the test beam propagating through the cell and the reference beam propagating through the reference cell; and determining the mass of one or more cells using the following equation:
[0053]
[0054] where m is the cell mass, α is a constant describing the relationship between phase shift and cell mass, is the measured fractional phase shift, λ is the illumination wavelength, and is integrated over the total cell area A.
[0055] Metho...
Embodiment 1
[0076] Example 1: Exemplary Methods and Materials Useful for Embodiments of the Invention
[0077] Interferometer
[0078] Live cell interferometers have been described in detail previously (1). Briefly, the system is an optical microscope based on a modified Veeco NT9300 optical profiler equipped with a 20X0.28NA Michelson interference objective lens, which can not only ) observes side features and can observe the height dimension of reflective targets down to 1 nanometer scale. A Michelson interferometer consists of a beam splitter, a reference mirror, and a compensating fluid cell to adjust for differences in the optical path caused by the fluid surrounding the sample. The phase shift interferometry (PSI) method was used to obtain the phase diagram of the cell body in situ (14). During the measurement, the piezoelectric transducer slightly reduces the optical path (optical path), which causes a phase shift between the test beam and the reference beam. The system records...
Embodiment 2
[0138] Example 2: Quantification of real-time drug sensitivity of single and clustered breast cancer cells by mass profiling
[0139] As mentioned above, viable cell mass profiling is a promising new approach to rapidly quantify responses to therapeutic agents through picogram-scale changes in cell mass over time. A significant obstacle to mass analysis is the inadequacy of existing methods to deal with polymorphic cell clusters and clumps, which are more commonly present in disease-derived samples or tissue cultures rather than isolated single cells. Herein, an automated live cell interferometer (aLCI) is provided as a single cell and colony-forming human breast cancer cell line against HER2-directed monoclonal antibody (HER2-directed monoclonal antibody), trenstizumab (Herceptin). Evidence of sensitivity for fast and accurate quantifiers. The relative sensitivity was determined to be tens to hundreds of times faster than feasible conventional proliferation assays. These aL...
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