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Confocal laser optical tweezers Raman spectroscopy test device capable of being used in upright and inverted manners

An optical tweezers Raman and spectral testing technology, which is applied in the field of cell or organelle research, can solve the problems of high laser power, inability to make full use of the numerical aperture of the oil lens lens, and the inability of cells to be captured by optical tweezers.

Active Publication Date: 2014-01-08
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the international research groups have built up inverted laser optical tweezers Raman systems, but it has some shortcomings: the laser needs to go through the oil lens oil and the cover glass to grasp the cells from the objective lens to the cells. However, the spectral signal tested in this way is easily interfered by the background signal of the immersion oil and the cover glass. At the same time, the numerical aperture of the oil immersion lens cannot be fully utilized, and the captured cells can only be limited to 15 cells above the cover glass. Cells around microns cannot be captured by optical tweezers at other locations, which brings great inconvenience to the actual operation; although the upright optical tweezers system can extend the lens of the water mirror to any depth in the solution for cell capture, the water mirror cannot The numerical aperture of the lens is relatively small, and a larger laser power is required when using an upright system to capture cells, so it is difficult to achieve the optical tweezers capture of large-mass cells or other particles

Method used

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  • Confocal laser optical tweezers Raman spectroscopy test device capable of being used in upright and inverted manners

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Experimental program
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Effect test

Embodiment 1

[0017] First, the single-mode semiconductor Raman laser outputs 785nm laser light, which is reflected by mirror A 1 and mirror B 23 and then passes through convex lens A 22, pinhole 21, convex lens B 20, band-pass color filter 19, mirror C 18 and reflector After the mirror D 3, the laser light is incident on the 785nm short-wave pass dichroic beam splitter 4 with an incident angle of 45° and is transmitted through the beam splitter, and the laser light is introduced into a reflective mirror E 5 equipped with a 45° reflection angle to reflect and enter Upright the microscope 11 until the 63 times Leica water mirror lens in the upright microscope 11, the emitted laser captures the active cells in the cell culture dish A 12 under the water mirror, and the laser power is about 10mw at this time. Use the built-in CCD in the microscope to observe whether the cells are stably trapped in the potential well formed by the laser focus. When the cell is trapped in the optical tweezers wel...

Embodiment 2

[0020] Turn the upper reflector E5 in the upright microscope 11 to move out of the optical path, so that the laser light is directly emitted to the reflector F6, and then reflected by the reflector G10 and the reflector H9, the laser light is introduced into the inverted microscope 8. The 100x oil immersion lens with a high numerical aperture configured in the microscope 8 can more easily realize the optical tweezers capture and Raman spectrum testing of cells.

[0021] The above-mentioned laser is a 785nm single-mode semiconductor Raman laser (manufacturer: Shanghai Xilong Optoelectronics Technology Co., Ltd., model: SL-785-XXX), and the maximum power of the emitted laser is 100mW; the upright microscope is Leica brand; Kaiser spectrometer (model: HoloSpec-f_2.2-NIR); mirror model: PF10-03-P01-10 spatial filter (KT310), convex lens (model: LA1608-B-ML), bandpass filter (model: FL780-10) , dichroic beam splitter (LPD01-785RU-25), notch filter (NF01-785U-25), purchased from Tho...

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Abstract

The invention relates to a confocal laser optical tweezers Roman spectroscopy test device capable of being used in upright and inverted manners. The confocal laser optical tweezers Roman spectroscopy test device is composed of a semiconductor Roman laser, a spatial filter, a short-pass dichroic beam-splitting optical path system, a spectrometer, an upright microscope and an inverted microscope. The laser is connected with the spatial filter in an optical path manner. The spatial filter is connected with the short-pass dichroic beam-splitting optical path system in an optical path manner. The short-pass dichroic beam-splitting optical path system is connected with the upright microscope in an optical path manner. The short-pass dichroic beam-splitting optical path system is connected with a Roman detector in an optical fiber manner. The short-pass dichroic beam-splitting optical path system is connected with the upright microscope through a reflector in the optical path manner. The short-pass dichroic beam-splitting optical path system is connected with the inverted microscope through other reflectors in an optical path manner. By the test device, activity of cells can be guaranteed, and biochemical analysis of the cells can be performed. Upright and inverted laser optical tweezers Roman spectroscopy systems which can be switched mutually, flexibly and conveniently through a rotary table with a 45-degree reflector can be used in a combined manner or independently.

Description

technical field [0001] The invention relates to a confocal laser optical tweezers Raman spectrum testing device that combines upright and inverted, which can realize optical tweezers capture and Raman spectrum testing of micron-scale transparent substances, and is especially suitable for cell or organelle research . By analyzing the obtained Raman spectral data, it is possible to distinguish and identify changes in the microstructure or biochemical composition of biological cells. Background technique [0002] The laser optical tweezers Raman system is an optical technology that combines laser optical tweezers with Raman spectroscopy. It is a popular tool for the study of single-cell molecules in the biological field today. It has the characteristics of non-destructive, fast, and real-time detection. At present, most of the international research groups have built up inverted laser optical tweezers Raman systems, but it has some shortcomings: the laser needs to go through t...

Claims

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Application Information

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IPC IPC(8): G01N21/65G01N21/01
Inventor 冯尚源陈荣李永增黄少华林居强黄组芳
Owner FUJIAN NORMAL UNIV
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