CN210571972U

CN210571972U - micro-Raman spectrometer

Info

Publication number: CN210571972U
Application number: CN201921046642.XU
Authority: CN
Inventors: 徐艇
Original assignee: Wuxi Ruisheng Medical Technology Co ltd
Current assignee: JIANGSU RAYME BIOTECHNOLOGY Co.,Ltd.
Priority date: 2019-05-10
Filing date: 2019-07-06
Publication date: 2020-05-19
Anticipated expiration: 2029-07-06

Abstract

The utility model discloses a micro-Raman spectrometer, including exciting light source, exciting light source lower extreme is connected with fiber probe, and fiber probe is through the little system operation of control side-to-side movement system connection system operation a little, is provided with the little system operation of control up-and-down movement on the system operation a little, and fiber probe stretches into in the light shielding shell, is provided with the culture dish that is used for placing the cytoplasm in the light shielding shell, and the raman spectrometer is connected for inverting the imager in culture dish below, the below of inverting the imager. And the sample inversion imager is also connected with a mobile control system. The Raman spectrum of the micro-area such as the level of a single cell is collected to detect the material components in the micro-area such as the single cell, and the fingerprint of the chemical material of the whole single cell can be obtained without damage, so that the phylogeny, the physiological characteristics, the metabolite change and the like of the living single cell can be rapidly identified, and the method has important significance for the function identification and the resource development of the microorganism which is difficult to culture.

Description

micro-Raman spectrometer

Technical Field

The utility model relates to a micro spectrometer belongs to the applied technical field of optical technology in biological field.

Background

Compared with fluorescence spectrum, infrared spectrum and other technologies, the Raman scattering technology has the advantages of basically no disturbance to living cells under the condition of no need of fluorescence labeling, no need of labeling for in-situ measurement and no need of preparation, and the Raman spectrum is not disturbed by water. Therefore, the Raman spectrum technology has outstanding advantages compared with the research and the determination of living cells, especially the continuous monitoring of single living cells.

The Raman spectrum technology is used for carrying out nondestructive analysis on the sample, and the method has the characteristics of non-contact and nondestructive testing of the sample, high detection sensitivity, short time, small sample amount, no need of preparation of the sample and the like.

The single cell sorting method based on the Raman microscopic spectrum technology does not need additional marks, and can nondestructively obtain the chemical substance fingerprint of the whole single cell, thereby rapidly identifying the phylogeny, the physiological characteristics, the metabolite change and the like of the living single cell, and having important significance for the function identification and the resource development of microorganisms difficult to culture.

At present, the Raman detector can only be used for macroscopic detection and can not realize single cell level detection.

Disclosure of Invention

The utility model aims to solve the technical problem of providing a micro-Raman spectrometer capable of detecting single cells.

The technical scheme of the utility model is that:

the utility model provides a micro-Raman spectrometer, includes the exciting light source, and exciting light source lower extreme is connected with fiber probe, and fiber probe connects the system of operating a little through controlling the side-to-side movement system of operating a little, is provided with the system of operating a little that moves about the control on the system of operating a little, and fiber probe stretches into in the light shielding shell, is provided with the culture dish that is used for placing the cytoplasm in the light shielding shell, and the culture dish below is the inversion imager, and Raman spectrometer is connected to the below of inversion imager.

And the sample inversion imager is also connected with a mobile control system.

The utility model has the advantages that:

the micro-area detection can be realized: the sample detection in the 500nm-400um area can be realized through the nano-level optical fiber probe; the precise control of the position of the detected optical fiber probe is realized through a precise micro-operation system, so that the position precision of micro-area detection is ensured; micro Raman detection: the traditional Raman spectrometer is too large in size, expensive in price and very limited in detection position, and the latest miniature optical fiber Raman spectrometer has the characteristics of small size, high sensitivity, low price, flexible detection position and the like; the sample image and raman signal were performed simultaneously: the function of synchronously imaging the sample and detecting the Raman signal is realized through the inverted imager, the excitation light source and the micro Raman spectrometer. The Raman spectrum of the micro-area such as the level of a single cell is collected to detect the material components in the micro-area such as the single cell, and the fingerprint of the chemical material of the whole single cell can be obtained without damage, so that the phylogeny, the physiological characteristics, the metabolite change and the like of the living single cell can be rapidly identified, and the method has important significance for the function identification and the resource development of the microorganism which is difficult to culture.

Drawings

Fig. 1 is a schematic structural diagram of the present invention;

fig. 2 is a detection data diagram according to an embodiment of the present invention.

In the figure: 1-excitation light source; 2-optical fiber probe; 3-micro operating system; 3.1-controlling the micro-operation system of up-and-down movement; 3.2-controlling the left and right movement micro-operation system; 4-culture dish; 5-a movement control system; 6-inverting the imager; 7-cytoplasm; 8-an organelle; 9-Raman spectrometer; 10-a light shielding housing.

Detailed Description

The invention is further described with reference to the following specific drawings and examples.

The utility model provides a micro-Raman spectrometer, including exciting light source 1, exciting light source 1 lower extreme is connected with fiber probe 2, fiber probe 2 connects the micro-manipulation system 3 through controlling the side-to-side movement micro-manipulation system 3.2, be provided with on the micro-manipulation system 3 and control the up-and-down movement micro-manipulation system 3.1, fiber probe 2 stretches into in the light shielding shell 10, be provided with the culture dish 4 that is used for placing cytoplasm 7 in the light shielding shell 10, the culture dish 4 below is for inverting imager 6, Raman spectrometer 9 is connected to inverting imager 6's below, Raman spectrometer 9 carries out Raman signal collection through being connected to inverting imager 6.

The inverted imager 6 is also connected with a mobile control system 5, the mobile control system 5 adjusts and controls the relative position, and when Raman detection is carried out, the light shielding shell is completely closed, so that interference of an external light source on detection is avoided.

The fiber probe 2 is divided into two types: a large exciting light probe 12 with head size in 100-900nm for detecting subcellular level can be extended into cell and nucleus for excitation; another small excitation light probe 2, with a head size in the range of 1-500um, is used to detect the level of whole single cells,

according to the invention, the light source required by the excitation light source 1 is used for exciting the cytoplasm 7 on the culture dish 4 through the micro-nano detection probe (the detector is accurately positioned through controlling the adjusting device by the micro-nano detection probe), and the generated result is received, analyzed and processed by the Raman spectrometer 9 to obtain the required result.

The excitation light source 1 can generate excitation light with any single wavelength within the range of 230-850nm, and can also be selected by a built-in monochromator, the wavelength of the excitation light source is 230-, and the excitation light source can be manufactured by ThorLabs company, USA, model: SAMBA 400; the optical fiber probe 2 can be made of PMA-P-O, model number, produced by Jiangsu Ruiming biology company; the micro-operation system 3 can be produced by Jiangsu Ruiming biological company, and has the following model: AM 01; the micro-operation system 3.1 for controlling the up-and-down movement can be produced by Jiangsu Ruiming biology company, and has the following model: AM 01-UD; the left and right movement control micro-operation system 3.2 can be produced by Jiangsu Ruiming biology company, and has the following model: AM 01-LR; the mobile control system 5 can be manufactured by Jiangsu Ruiming biological company, model number: AW 01; the inverted imager 6 may be manufactured by olympus corporation of japan, model number: IX 83; the micro raman spectrometer 9 can be produced by china such as the maritime optical company, and has the following model: SEED 3000.

Example (b):

experimental materials: the sample was HeLa cells (human cervical cancer cells, manufactured by Cobioer Co., Ltd., cat # CBP 60232); light source 1: wavelength 785nm, raman spectrometer 9: the resolution ratio is optimally up to 4 cm; moving micro operation 3: minimum precision 10 um; the optical fiber probe 2: the diameter of the optical fiber core is 150 nm; inverting the imager 6: the online imaging function can be realized; the culture dish 4: 5mm thick, 8cm diameter.

The experimental process comprises the following steps:

the sample cells adhere to the bottom of the culture dish (before the experiment, the culture dish is filled with phosphate buffered saline-PBS and is placed in a carbon dioxide incubator for 12 hours at 20 ℃), and the culture dish is positioned in the middle of the stage of the inverted microimager; the nanometer optical fiber detection probe 2 is precisely adjusted to a region with lower cell distribution density by moving the micro-manipulator 3 and the movement control system 5, and then the fiber core is inserted into the position by the depth of about 100um by adjusting the Z axis (the depth can be controlled by the micro-manipulator); and (3) opening an excitation light source in the searched area, adjusting the power to 3mW, simultaneously opening the fiber Raman spectrometer 9, and after a stabilization period of 10 seconds, starting the spectrometer to receive signals.

The experimental results are as follows:

the detected wave crests 1-4 are respectively 1.C-O, 2. C-N3. C-H4. RC = CR' bonds in the cells, and the peak size reflects the abundance of various groups in the cells.

Claims

1. A micro Raman spectrometer is characterized in that: the Raman spectrometer comprises an excitation light source (1), wherein the lower end of the excitation light source (1) is connected with an optical fiber probe (2), the optical fiber probe (2) is connected with a micro-manipulation system (3) through a control left-right movement micro-manipulation system (3.2), the micro-manipulation system (3) is provided with a control up-down movement micro-manipulation system (3.1), the optical fiber probe (2) extends into a light shielding shell (10), a culture dish (4) used for placing cytoplasm (7) is arranged in the light shielding shell (10), an inverted imager (6) is arranged below the culture dish (4), and a Raman spectrometer (9) is connected below the inverted imager (6).

2. The micro-raman spectrometer of claim 1, wherein: the inverted imager (6) is also connected with a mobile control system (5).