Novel calibration device and calibration method for total internal reflection fluorescence microscope penetration depth

A fluorescence microscope and total internal reflection technology, applied in the field of fluorescence microscope imaging, can solve the problems of inconvenient use and achieve the effects of simple operation, high calibration efficiency and simple device

Inactive Publication Date: 2017-08-04
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] Although these two methods can obtain better calibration results, they require special fluorescent beads or experimental tools, and complex image processing and data fitting are required in the later stage, which is not very convenient to use.

Method used

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  • Novel calibration device and calibration method for total internal reflection fluorescence microscope penetration depth
  • Novel calibration device and calibration method for total internal reflection fluorescence microscope penetration depth
  • Novel calibration device and calibration method for total internal reflection fluorescence microscope penetration depth

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Embodiment 1

[0054] The calibration device for the incident depth of the total internal reflection fluorescence microscope established above is placed in the scanning galvanometer system of Thornlabs, and then the laser is emitted from the laser produced by Melles Griot, and then imported into the collimating lens of Thornlabs through a single-mode fiber to complete the collimation ; The collimated light beam is incident into the scanning galvanometer system of Thornlabs Company for optical path deflection, and then incident into the scanning lens of Thornlabs Company for focusing. The light beam emitted by the scanning lens is expanded by the first field mirror, and then exits in parallel, and after being focused and reflected by the focusing lens and dichroic mirror of Thornlabs, the light beam is focused on the objective lens of the total internal reflection fluorescence microscope produced by OLYMPUS and projected on it. on the sample stage. The signal light emitted by the sample to be...

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Abstract

The invention discloses a calibration device for the total internal reflection fluorescence microscope penetration depth. The device comprises a glass slide, square cover glass and a glass sheet, wherein the square cover glass covers the upper surface of the glass slide; the right sides of the square cover glass and the glass slide are aligned; the glass sheet is arranged on the glass slide; the left side edges of the glass sheet and the glass slide are in contact and are aligned; the lower surface of the glass sheet is supported by the direction cover glass; the lower surface of the glass sheet is uniformly coated with fluorescent dye. The invention also discloses a method for calibrating the total internal reflection fluorescence microscope penetration depth by using the calibration device. The device has the advantages that the structure is simple; the cost is low; the operation is convenient and flexible.

Description

technical field [0001] The invention belongs to the field of fluorescence microscope imaging, and in particular relates to a novel calibration device and calibration method for the incident depth of a total internal reflection fluorescence microscope. Background technique [0002] Total internal reflection fluorescence microscopy (TIRFM) is an emerging optical imaging technology in recent years. Very thin regions of the interface. Due to the characteristic of the evanescent wave field that the energy decays exponentially in the axial direction, the dynamic range of the observable area is usually below 200nm, which effectively controls the excitation volume and greatly reduces the interference of background light noise, so it has other optical imaging technologies. Unmatched high signal-to-noise ratio and contrast. Therefore, this technology has been widely used in the dynamic observation of substances on the surface of cell membranes. [0003] According to the characteris...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N21/01G02B21/00G02B21/34
CPCG01N21/6456G01N21/01G01N2021/0112G02B21/0076G02B21/34
Inventor 许迎科金璐红刘旭
Owner ZHEJIANG UNIV
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