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

A microscope for use in total internal reflection fluorescence microscopy (TIRFM) is provided. The microscope includes a first white-light source for directing light along a first optical path; an annular slit member disposed in the first optical path, the annular slit member having an annular slit for blocking all but an annulus of light corresponding to the annular slit; and an objective lens for directing the annulus of light to a specimen such that TIRFM of the specimen is achieved. Also provided are various ways for converting the microscope to and from a TIRFM microscope and a conventional microscope.

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 total internal reflection fluorescence microscope capable of being used with an atomic force microscope and belongs to the field of optical microscopes. The fluorescence microscope comprises a support assembly, an imaging assembly, a base, a sample objective table of the atomic force microscope. The size of the support assembly can be adjusted according to the size of the sample objective table. In the imaging assembly, a bearing body is arranged on the base, a one-dimensional translation seat is arranged on the bearing body, a second connection seat is fixed on the bearing body, a lens cone lens is arranged in an inner cavity and a reflection mirror is arranged on the bearing body and under the lens cone lens. A lower base plate of a first connection seat of a U-type structure is fixed on the one-dimensional translation seat. A two-dimensional translation seat is fixed on the outer end of a left side arm of the first connection seat. A fiber coupler is arranged in the two-dimensional translation seat in a two-dimensional moving manner. The end part of a first lens and the end part of a second lens are arranged on two ends of a second through hole of the right side arm of the first connection seat. A dichroscope assembly is arranged on the second connection seat. An objective lens is fixed on the dichroscope. The objective lens is arranged in an objective lens installation seat in a height-adjustable manner.

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.

The invention discloses a method for detecting the laterally limited area of ​​a living plant cell membrane protein. The method comprises: (1) expressing a fusion protein in a plant to obtain a transgenic plant material with single-color fluorescent labeling for the protein to be tested; (2) observing the transgenic plant material with a variable-angle total internal reflection fluorescence microscope, Obtain time series images; (3) use single particle tracking technology to identify and track fluorescent points in time series images, obtain the trajectory of fluorescent points over time, and then calculate the average square displacement of fluorescent points over time; (4) use The Origin software fits the average square displacement of the fluorescent point over time, obtains the corresponding constant, and calculates the moving range of the fluorescent point. The method for calculating the laterally restricted area of ​​membrane proteins in living cells is simple and repeatable, and individual information of proteins and group average information can also be calculated.