35 results about "Total internal reflection fluorescence microscope" patented technology

Techniques are provided for illuminating a sample by using total internal reflection (TIR) from a diffraction limited focused annular illumination beam. The illumination forms an affected region and an overlapping confocal region that may have dimensions the below 1 μm. An adjustable diffractive optical element, for example, may create a second order Bessel profile laser beam that is focused on a sample using a high-numerical aperture objective under TIR. An evanescent field excites fluorescent biological material in the confocal region, and fluorescence from the material is analyzed in fluorescence correlation spectroscopy system.

Correction elements that can be incorporated in objective-based TIRF microscopy instruments to correct for chromatic aberrations are described. A correction element can be placed between a multiple wavelength excitation beam source and the microscope objective lens. In one aspect, the thickness of the correction element is defined to compensate for different axial positions of the focal points associated with each excitation wavelengths traveling along the outer edge of lenses comprising a microscope objective lens. In another aspect, the correction element can be angled and / or configured so that the different wavelengths of multiple wavelength excitation light are shifted to adjust the angle of incidence for each wavelength at the specimen / substrate interface.

A microscope apparatus has an illumination optical system illuminating a sample with laser light from laser light sources. A fluorescence detection optical system detects fluorescence from the sample. Fluorescence cubes are interchangeably provided in an optical path of the illumination optical system and lead the laser light to the sample. An objective lens is also provided. At least one of the fluorescence cubes includes an optical member that makes a principal ray of the laser light substantially parallel to an optical axis of the illumination optical system and concentrates the laser light on a given position that is on a pupil position of the objective lens and separated from the optical axis, thereby providing a microscope apparatus capable of changing from a confocal microscope to a total-internal-reflection fluorescence microscope by exchanging a fluorescence cube used in the fluorescence microscope.

The invention discloses a detection method of a trace amount of a target based on monomolecular fluorescent sensing. The detection method comprises the following steps of: (1) preparing a standard solution containing a target object; (2) fixing an auxiliary probe on a slide, adding an aptamer, and hybridizing the auxiliary probe with the aptamer; (3) adding the standard solution to the slide, andbinding the target object with the aptamer to separate the target object from the auxiliary probe; (4) adding a signal probe to the slide, and carrying out transient hybridization on the signal probeand the released auxiliary probe; (5) placing the slide on an object stage of a total internal reflection fluorescence microscope for detection, recording single-molecule fluorescence points with thenumber of times of change of the fluorescence state being more than 15 times, taking the single-molecule fluorescence points as an ordinate and taking the concentration of the target object in the standard solution as an abscissa to make a standard curve; and (6) repeating the steps (3) to (5) for a sample to be detected, recording single-molecule fluorescence points with the number of times of change of the fluorescence state being more than 15 times, and obtaining the concentration of the target object in the sample to be detected according to the standard curve.

An inverted microscope system includes an objective lens holding unit that holds an objective lens configured to collect at least observation light from a specimen, a tube lens configured to form an image using the observation light collected by the objective lens, a total internal reflection fluorescence microscopy optical system provided between the objective lens and the tube lens and configured to observe the observation light from the specimen using a total reflection illumination, and a disk scanning confocal optical system including a rotary disk on which a confocal opening is formed, the confocal opening being placed at a position substantially conjugate to a focus position of the objective lens. A relative distance between the focus position of the objective lens and the substantially conjugate position is changeable along an optical path of the observation light.

The invention discloses a method for synchronously monitoring chemical signals and electrical signals of nerve cell networks. Firstly, the nerve cell network is cultivated in situ on a pretreated microelectrode array; then, the neuron vesicles are dyed, and then buffer solution is added ; Observing the nerve cell network with a total internal reflection fluorescence microscope, connecting the shooting camera and the controller of the microelectrode array with the pulse signal generator; performing external stimulation on the nerve cell network, and simultaneously turning on the pulse signal generator, shooting the camera and the microelectrode array The controller collects the fluorescent signal and the electrical signal synchronously, respectively. The synchronous monitoring method of the present invention has low background signal, high signal-to-noise ratio, and has space-time resolution responsiveness, which can meet the monitoring requirements of nerve cell networks; the method has stable signals, high sensitivity, and can realize imaging measurement for various nerve cells and networks, and High-throughput analysis can be achieved.