CN109374487A - A kind of Ultrafast spectrum research device of microparticle - Google Patents
A kind of Ultrafast spectrum research device of microparticle Download PDFInfo
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- CN109374487A CN109374487A CN201811220428.1A CN201811220428A CN109374487A CN 109374487 A CN109374487 A CN 109374487A CN 201811220428 A CN201811220428 A CN 201811220428A CN 109374487 A CN109374487 A CN 109374487A
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- 239000011859 microparticle Substances 0.000 title claims abstract description 50
- 238000001228 spectrum Methods 0.000 title claims abstract description 24
- 238000011160 research Methods 0.000 title claims abstract description 11
- 238000003756 stirring Methods 0.000 claims abstract description 73
- 238000006073 displacement reaction Methods 0.000 claims abstract description 41
- 230000003287 optical effect Effects 0.000 claims abstract description 14
- 238000005086 pumping Methods 0.000 claims description 15
- 238000001514 detection method Methods 0.000 claims description 8
- 229920001343 polytetrafluoroethylene Polymers 0.000 claims description 8
- 239000004810 polytetrafluoroethylene Substances 0.000 claims description 8
- -1 polytetrafluoroethylene Polymers 0.000 claims description 4
- 229910001220 stainless steel Inorganic materials 0.000 claims description 3
- 239000010935 stainless steel Substances 0.000 claims description 3
- 238000000034 method Methods 0.000 abstract description 19
- 239000007788 liquid Substances 0.000 abstract description 8
- 238000005259 measurement Methods 0.000 abstract description 6
- 238000002474 experimental method Methods 0.000 abstract description 5
- 238000001917 fluorescence detection Methods 0.000 abstract description 2
- 239000000463 material Substances 0.000 abstract description 2
- 238000010586 diagram Methods 0.000 description 16
- 239000012530 fluid Substances 0.000 description 13
- 239000007850 fluorescent dye Substances 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 230000000737 periodic effect Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000000886 photobiology Effects 0.000 description 1
- 238000004080 punching Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/06—Investigating concentration of particle suspensions
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/06—Investigating concentration of particle suspensions
- G01N15/075—Investigating concentration of particle suspensions by optical means
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- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The present invention relates to material engineering fields, a kind of Ultrafast spectrum research device of microparticle, including displacement platform I, rotation motor, rotary shaft, magnet, displacement platform II, sample cavity, stirring sheet, lens I, beam splitter, laser, optical filter, lens II, aperture, lens III, detector and computer, the microparticle sample of low concentration is studied using Ultrafast spectrum method, with special sample cavity and stirring sheet, the sample of different zones in sample cavity is enabled to carry out fast exchange, meet the needs of Ultrafast spectrum experiment, high-precision measurement is carried out to the quantity of microparticle in liquid using the method for rotation sample cavity combination fluorescence detection, sample loss is not will cause, the sample of different zones is able to carry out fast exchange in sample cavity, and noise is small, high precision count can be carried out to microparticle sample, no sample loss , operating process simplicity.
Description
Technical field
The present invention relates to material engineering field, especially a kind of microparticle that low concentration is studied using Ultrafast spectrum method
A kind of Ultrafast spectrum research device of microparticle of sample.
Background technique
Ultrafast spectrum be it is a kind of using ultrafast laser come the method for study sample characteristic, can obtain with femtosecond resolution ratio
Time-resolved spectrum, the laser for needing to focus commonly used in the ultrafast process in research photochemistry or photobiology, in experiment
It is mapped to the partial region in sample, the laser repetition rate of the laser system of the prior art can reach kHz magnitude, this just needs sample
There is sufficiently fast exchange velocity between the part of laser excitation and unawakened part in product, to avoid sample by laser
Repeat excitation and experimental result impacted, certain prior arts using the method for circulating pump come so that part in sample into
Row exchange, the sample size not only needed is larger, and can introduce mechanical noise, or even will cause specimen breakdown；Other are existing
Technology carries out the sample exchange of different zones using the blender at the uniform velocity rotated, but in the sample cavity of certain small volumes
It is smaller to stir range, i.e., can only be stirred mixing to the sample around stirring sheet, and sample can be made to generate laser
Periodically refraction, causes to introduce periodic noise in Ultrafast spectrum, have an impact to the result of Ultrafast spectrum.Certain
It in, needs to count the numbers of particles in liquid, count measurement in the prior art takes a long time, and can only be to list
The biggish sample of amounts of particles in the volume of liquid of position measures, and a kind of Ultrafast spectrum research device of microparticle can
It solves the problems, such as.
Summary of the invention
To solve the above-mentioned problems, apparatus of the present invention have special sample cavity and stirring sheet, enable in sample cavity
The sample of different zones carries out fast exchange, meets the needs of Ultrafast spectrum experiment, in addition, using rotation sample cavity combination fluorescence
The method of detection carries out high-precision measurement to the quantity of microparticle in liquid, not will cause sample loss, easy to operate.
The technical scheme adopted by the invention is that:
A kind of Ultrafast spectrum research device of microparticle includes displacement platform I, rotation motor, rotary shaft, magnet, displacement
Platform II, sample cavity, stirring sheet, lens I, beam splitter, laser, optical filter, lens II, aperture, lens III, detector and meter
Calculation machine, xyz are three-dimensional coordinate system, and rotation motor is fixed on displacement platform I, the laser, beam splitter, lens I and sample
Product chamber successively forms input path, the sample cavity, lens I, beam splitter, optical filter, lens II, aperture, lens III and detection
Device successively forms emitting light path, laser can the adjustable continuous laser of launch wavelength and laser pulse, laser pulse includes pump
Pu pulse and direct impulse, the pumping pulse and direct impulse have different power, frequency and duration, direct impulse
Time interval between pumping pulse can be adjusted；Magnet is parallel to z coordinate by rotation axis connection rotation motor, rotary shaft
Axis, displacement platform I can be three-dimensional mobile, and rotation motor is able to drive magnet rotation, and sample cavity is fixed on displacement platform II, displacement platform
II can be three-dimensional mobile, and displacement platform II is able to drive sample cavity and rotates around the y direction center line of sample cavity, sample cavity top half
Be rectangular body cavity, lower half portion be the cylinder barrel of axis in the y-direction, between the top half and lower half portion have aperture plate,
The rectangular body cavity is 30 millimeters a height of, 10 millimeters a length of, width is 2 millimeters, and the cylinder barrel is 20 millimeters a height of, basal diameter 10
Millimeter, the y direction center line of the rectangular body cavity and the axis collinear of the cylinder barrel, stirring sheet is by having magnetic stainless steel
It wraps up polytetrafluoroethylene (PTFE) outside bar to be made, the diameter of stirring sheet is 1.2 millimeters, length is 13 millimeters, and stirring sheet is located on sample cavity
Half part, the aperture plate can prevent stirring sheet from falling into cylinder barrel；The rotation motor, laser and detector difference cable connect
Computer is connect, the time between the laser pulse of laser sending and rotation motor rotation angle can be adjusted by computer
Difference；The size of aperture can be adjusted, and the detection resolution of detector can be adjusted according to various sizes of microparticle to be measured.
Stirring sheet makes the principle of the sample part progress fast exchange of the different zones in sample cavity are as follows:
Adjusting displacement platform II makes the width direction of the rectangular body cavity of the top half of sample cavity parallel with z-axis, adjusts position
Moving stage I makes the axis extended line of rotary shaft be located at the center of rectangular body cavity, opens rotation motor and makes magnet around rotary shaft
Axis at the uniform velocity rotates, and revolving speed representative value is 0.5 revolutions per second, and due to the magnetic torque effect by magnet, stirring sheet will be revolved with magnet
Turn, stirring sheet is rotated up both ends and contacts and be limited with the inner wall of rectangular body cavity, and magnet continues to rotate at this time, stirring sheet still by
Magnetic torque to magnet acts on, but stirring sheet position is constant, when magnet is rotated to a certain angle, magnetic of the stirring sheet by magnet
Torque is reversed, and stirring sheet can quickly be overturn to the opposite direction that magnet rotates at this time, and stirring sheet is overturn until both ends and rectangular body cavity
Inner wall contact and be limited, in conclusion stirring sheet is rotated twice during magnet at the uniform velocity rotates a circle, once
It is stirring sheet as magnet direction of rotation rotates at a slow speed, is that the opposite direction that stirring sheet is rotated to magnet is quickly overturn again, stirs
The quick switching process for mixing piece makes the sample of stirring sheet near zone and the sample fast exchange of sample cavity other parts.Technology
Advantage is: stirring sheet will not introduce noise with the rotary course at a slow speed of magnet direction of rotation, by adjusting laser pulse and stirring
The time interval for mixing the quick switching process of opposite direction that piece is rotated to magnet, can be effectively prevented from mechanical noise in sample into
Enter spectrum, also avoids the periodic refraction of laser in the sample.
The principle of count measurement is carried out to the microparticle in fluid sample are as follows:
Fluid sample containing microparticle to be measured is placed in sample cavity, according to the type of microparticle to be measured, to fluid sample
Middle addition fluorescent dye enables the microparticle surfaces under the laser irradiation of specific wavelength to launch fluorescence, laser hair
The laser irradiation sample penetrated makes microparticle launch fluorescence, and the fluorescence is recorded by emitting light path by detector；Laser
The laser of transmitting is focused in the cylinder barrel of sample cavity lower half portion after the deviation of beam splitter by lens I, displacement platform II band
Dynamic sample cavity is rotated around the y direction center line of sample cavity, and the microparticle to suspend in fluid sample is moved together with solvent, and is passed through
The focus of sample endovenous laser is to launch fluorescence；In the non-rotary situation of sample cavity, wherein the part being irradiated with a laser
This partial volume that the fluorescence that microparticle in volume is launched can be detected by detector, is defined as detectable volume, estimates
The method for calculating detectable volume is as follows: the fluorescence reference for the fluorescence that will launch is fixed in sample cavity, and is made
Detector can identify that laser beam is along negative z direction incidence sample cavity, when sample cavity is fixed, detector record
The intensity of fluorescence signal is(formula one), wherein w0It is sharp
Beam waist diameter of the light beam in the direction x, z0For the beam waist diameter of laser beam in the z-direction, adjusts displacement platform II and make sample cavity along the side x
To translation, detector is recorded the fluorescence signal of fluorescence reference, obtains w after being fitted using dimensional gaussian distribution to data0,
Adjusting displacement platform II translates sample cavity in the z-direction, and detector records the fluorescence signal of fluorescence reference, using dimensional Gaussian
Distribution obtains z after being fitted to data0, it is fitted using formula a pair of the data obtained, estimates detectable volume, representative value
It is 0.5 nanoliter；Detector continuously records the intensity for the fluorescence that the microparticle in a period of time T in detectable volume is launched, and obtains
To the signal data of some varying strengths, computer obtains detectable volume in T time after handling the signal data
The quantity of interior microparticle, and the revolving speed of sample cavity is combined, the quantity of microparticle in unit liquid volume can be obtained.
The step of Ultrafast spectrum experiment is carried out to microparticle sample using a kind of Ultrafast spectrum research device of microparticle
Suddenly are as follows:
Step 1, the fluid sample containing microparticle to be measured is placed in sample cavity；
Step 2, adjusting displacement platform II makes the width direction of the rectangular body cavity of the top half of sample cavity and z coordinate axis flat
Row adjusts the center for the rectangular body cavity that displacement platform I makes the axis extended line of rotary shaft be located at sample cavity top half；
Step 3, the position of lens I, beam splitter, laser, optical filter, lens II, aperture, lens III and detector is adjusted
It sets, so that the laser of laser transmitting is focused to the cuboid of sample cavity top half by lens I after the deviation of beam splitter
In chamber, and make after passing sequentially through lens I, beam splitter, optical filter, lens II, aperture and lens III by the light of sample reflection,
Into detector；
Step 4, open rotation motor magnet is at the uniform velocity rotated around the axis of rotary shaft, revolving speed representative value be 0.5 turn/
Second, so that stirring sheet is stirred sample；
Step 5, laser periodically launches pumping pulse and direct impulse sequence, pumping pulse and direct impulse week
It is incident in sample to phase property；
Step 6, it according to the rotation angle of the corresponding rotation motor in stirring sheet position, is issued by computer regulated laser
Pumping pulse and direct impulse interval time so that the quick switching process of opposite direction that stirring sheet is rotated to magnet is pumping
Pulse and direct impulse are incident in the time slot of sample；
Step 7, detector records the light projected from sample and generates corresponding data, and computer obtains after analyzing the data
To the Ultrafast spectrum of sample.
Quick counter measurement is carried out to the microparticle in fluid sample using a kind of Ultrafast spectrum research device of microparticle
The step of are as follows:
Fluid sample containing microparticle to be measured is placed in sample cavity by step 1；
Step 2 adds fluorescent dye into fluid sample according to the type of microparticle to be measured, so that in respective wavelength
Laser irradiation under microparticle surfaces can launch fluorescence；
Step 3 adjusts the position of lens I, beam splitter, laser, optical filter, lens II, aperture, lens III and detector
It sets, so that the laser of laser transmitting is focused to the cylinder barrel of sample cavity lower half portion by lens I after the deviation of beam splitter
In, and make after passing sequentially through lens I, beam splitter, optical filter, lens II, aperture and lens III by the light of sample reflection, into
Enter detector；
Step 4, displacement platform II drive sample cavity to rotate around the y direction center line of sample cavity, and revolving speed a typical range of from 50 arrives
400 revs/min；
Step 5, the continuous laser that laser is launched are incident on the sample in sample cavity, the wave of the continuous laser
Length is determined according to the type of the fluorescent dye added in fluid sample；
Step 6 adjusts the size of aperture, and detector is enabled clearly to record the fluorescence of single microparticle transmitting；
Step 7, detector records the light projected from sample and generates corresponding data, after computer analyzes the data,
The quantity information for obtaining the microparticle sample to be measured in the detectable volume of fluid sample can be obtained in conjunction with the revolving speed of sample cavity
The quantity of microparticle in unit liquid volume, it is micro- in the unit volume liquid that can detect for 2 microns of microparticle of diameter
The minimum value of the quantity of particle is 50/milliliter.
The beneficial effects of the present invention are:
The sample of different zones is able to carry out fast exchange in sample cavity of the present invention, and noise is small, meets Ultrafast spectrum reality
The needs tested, in addition, high precision count, no sample loss, operating process simplicity can be carried out to microparticle sample.
Detailed description of the invention
It is further illustrated below with reference to figure of the invention:
Fig. 1 is schematic diagram of the present invention；
Fig. 2 is sample cavity side enlarged diagram；Fig. 3 is the top view of Fig. 2；
Fig. 4 is schematic diagram one of of the stirring sheet with magnet rotary course；
Fig. 5 is stirring sheet with the two of the schematic diagram of magnet rotary course；
Fig. 6 is stirring sheet with the three of the schematic diagram of magnet rotary course；
Fig. 7 is stirring sheet with the four of the schematic diagram of magnet rotary course；
Fig. 8 is stirring sheet with the five of the schematic diagram of magnet rotary course；
Fig. 9 is stirring sheet with the six of the schematic diagram of magnet rotary course.
In figure, 1. displacement platform I, 2. rotation motors, 3. rotary shafts, 4. magnet, 5. displacement platform II, 6. sample cavities, 7. stirrings
Piece, 8. lens I, 9. beam splitters, 10. lasers, 11. optical filters, 12. lens II, 13. apertures, 14. lens III, 15. detections
Device.
Specific embodiment
If Fig. 1 is schematic diagram of the present invention, xyz is three-dimensional coordinate system, including displacement platform I (1), rotation motor (2), rotation
Shaft (3), magnet (4), displacement platform II (5), sample cavity (6), stirring sheet (7), lens I (8), beam splitter (9), laser
(10), optical filter (11), lens II (12), aperture (13), lens III (14), detector (15) and computer, rotation motor
(2) it is fixed on displacement platform I (1), for magnet (4) by rotary shaft (3) connection rotation motor (2), rotary shaft (3) is parallel to z seat
Parameter, displacement platform I (1) can be three-dimensional mobile, and rotation motor (2) is able to drive magnet (4) rotation, and sample cavity (6) is fixed on position
On moving stage II (5), displacement platform II (5) can be three-dimensional mobile, and displacement platform II (5) is able to drive sample cavity (6) around sample cavity (6)
Y direction center line rotation, sample cavity (6) top half be rectangular body cavity, lower half portion be the cylinder barrel of axis in the y-direction, institute
Stating has aperture plate between top half and lower half portion, the rectangular body cavity is 30 millimeters a height of, 10 millimeters a length of, width is 2 millimeters,
The cylinder barrel is 20 millimeters a height of, basal diameter is 10 millimeters, y direction center line and the cylinder barrel of the rectangular body cavity
Axis collinear, by having magnetic stainless steel, package polytetrafluoroethylene (PTFE) is made stirring sheet (7) outside, and the diameter of stirring sheet (7) is
1.2 millimeters, length be 13 millimeters, stirring sheet (7) is located at sample cavity (6) top half, and the aperture plate can prevent stirring sheet (7)
Cylinder barrel is fallen into, the laser (10), beam splitter (9), lens I (8) and sample cavity (6) successively form input path, described
Sample cavity (6), lens I (8), beam splitter (9), optical filter (11), lens II (12), aperture (13), lens III (14) and detection
Device (15) successively forms emitting light path, laser (10) can the adjustable continuous laser of launch wavelength and laser pulse, laser arteries and veins
Punching includes pumping pulse and direct impulse, and the pumping pulse and direct impulse have different power, frequency and duration,
Time interval between direct impulse and pumping pulse can be adjusted, the rotation motor (2), laser (10) and detector
(15) difference cable connection computer can adjust the laser pulse and rotation motor of laser (10) sending by computer
(2) time difference between angle is rotated；The size of aperture (13) can be adjusted, can according to various sizes of microparticle to be measured come
Adjust the detection resolution of detector (15).
If Fig. 2 is sample cavity side enlarged diagram, if Fig. 3 is the top view of Fig. 2, sample cavity (6) top half is long
Cube chamber, lower half portion are the cylinder barrel of axis in the y-direction, y direction center line and the cylinder barrel of the rectangular body cavity
Axis collinear.
If Fig. 4 is schematic diagram one of of the stirring sheet with magnet rotary course, if Fig. 5 is stirring sheet with magnet rotary course
The two of schematic diagram, as Fig. 6 be stirring sheet with magnet rotary course schematic diagram three, if Fig. 7 is that stirring sheet is rotated through with magnet
The four of the schematic diagram of journey, as Fig. 8 be stirring sheet with magnet rotary course schematic diagram five, if Fig. 9 is that stirring sheet is revolved with magnet
The six of the schematic diagram of journey are turned over, arrow is direction of rotation in figure, and rotation motor (2) is able to drive magnet (4) rotation, magnet (4)
Position of magnetic pole change, stirring sheet (7) with magnet (4) rotate.
A kind of Ultrafast spectrum research device of the microparticle include displacement platform I (1), rotation motor (2), rotary shaft (3),
Magnet (4), displacement platform II (5), sample cavity (6), stirring sheet (7), lens I (8), beam splitter (9), laser (10), optical filter
(11), lens II (12), aperture (13), lens III (14), detector (15) and computer, xyz are three-dimensional coordinate system,
Rotation motor (2) is fixed on displacement platform I (1), the laser (10), beam splitter (9), lens I (8) and sample cavity (6) according to
Secondary composition input path, the sample cavity (6), lens I (8), beam splitter (9), optical filter (11), lens II (12), aperture
(13), lens III (14) and detector (15) successively form emitting light path, laser (10) can launch wavelength it is adjustable continuous
Laser and laser pulse, laser pulse include pumping pulse and direct impulse, and the pumping pulse and direct impulse have difference
Power, frequency and duration, the time interval between direct impulse and pumping pulse can adjust；Magnet (4) passes through rotation
Shaft (3) connects rotation motor (2), and rotary shaft (3) is parallel to z coordinate axis, and displacement platform I (1) can be three-dimensional mobile, rotation motor
(2) it is able to drive magnet (4) rotation, sample cavity (6) is fixed on displacement platform II (5), and displacement platform II (5) can be three-dimensional mobile,
Displacement platform II (5) is able to drive sample cavity (6) and rotates around the y direction center line of sample cavity (6), and sample cavity (6) top half is
Rectangular body cavity, lower half portion are the cylinder barrel of axis in the y-direction, have aperture plate, institute between the top half and lower half portion
State that rectangular body cavity is 30 millimeters a height of, 10 millimeters a length of, width is 2 millimeters, the cylinder barrel is 20 millimeters a height of, basal diameter is 10 millis
Rice, the y direction center line of the rectangular body cavity and the axis collinear of the cylinder barrel, stirring sheet (7) are magnetic stainless by having
It wraps up polytetrafluoroethylene (PTFE) outside steel pole to be made, the diameter of stirring sheet (7) is 1.2 millimeters, length is 13 millimeters, and stirring sheet (7) is located at
Sample cavity (6) top half, the aperture plate can prevent stirring sheet (7) from falling into cylinder barrel；The rotation motor (2), laser
(10) and detector (15) distinguishes cable connection computer, and the laser pulse of laser (10) sending can be adjusted by computer
With the time difference between rotation motor (2) rotation angle；The size of aperture (13) can be adjusted, can according to it is various sizes of to
Micrometer particle adjusts the detection resolutions of detector (15).
Stirring sheet (7) makes the principle of the sample part progress fast exchange of the different zones in sample cavity (6) are as follows:
Such as Fig. 1, adjusting displacement platform II (5) makes the width direction and z-axis of the rectangular body cavity of the top half of sample cavity (6)
In parallel, the center that displacement platform I (1) makes the axis extended line of rotary shaft (3) be located at rectangular body cavity is adjusted, rotation motor is opened
(2) rotate magnet (4) at the uniform velocity around the axis of rotary shaft (3), revolving speed representative value is 0.5 revolutions per second, such as Fig. 4, Fig. 5, due to by
Magnetic torque to magnet (4) acts on, and stirring sheet (7) will be rotated with magnet (4), and such as Fig. 6, stirring sheet (7) is rotated up both ends
It contacts and is limited with the inner wall of rectangular body cavity, such as Fig. 7, magnet (4) continues to rotate at this time, and stirring sheet (7) is still by magnet (4)
Magnetic torque effect, but stirring sheet (7) position is constant, such as Fig. 8, when magnet (4) rotation to a certain angle, stirring sheet (7) by
Magnetic torque to magnet (4) is reversed, and stirring sheet (7) can quickly be overturn to the opposite direction that magnet (4) rotate at this time, such as Fig. 9, stirring
Piece (7) is overturn until both ends contact and are limited with the inner wall of rectangular body cavity, in conclusion the process that magnet at the uniform velocity rotates a circle
In, stirring sheet (7) is rotated twice, is once stirring sheet (7) as magnet (4) direction of rotation rotates at a slow speed, is again
Stirring sheet (7) is quickly overturn to the opposite direction that magnet (4) rotates, and the quick switching process of stirring sheet (7) makes stirring sheet (7) attached
The sample of near field and the sample fast exchange of sample cavity (6) other parts.Technological merit is: stirring sheet (7) is with magnet (4)
The rotary course at a slow speed of direction of rotation will not introduce noise, be rotated by adjusting laser pulse and stirring sheet (7) to magnet (4)
The time interval of the quick switching process of opposite direction, the mechanical noise that can be effectively prevented from sample enter spectrum, also avoid
The periodic refraction of laser in the sample.
The principle of count measurement is carried out to the microparticle in fluid sample are as follows:
Fluid sample containing microparticle to be measured is placed in sample cavity (6), according to the type of microparticle to be measured, to liquid-like
Fluorescent dye is added in product, and the microparticle surfaces under the laser irradiation of specific wavelength is enabled to launch fluorescence, laser
(10) the laser irradiation sample emitted makes microparticle launch fluorescence, and the fluorescence is remembered by emitting light path by detector (15)
Record；The laser of laser (10) transmitting focuses to sample cavity (6) lower half by lens I (8) after the deviation of beam splitter (9)
In the cylinder barrel divided, displacement platform II (5) drives sample cavity (6) to rotate around the y direction center line of sample cavity (6), in fluid sample
The microparticle of suspension is moved together with solvent, and by the focus of sample cavity (6) interior laser to launch fluorescence；In sample cavity
(6) in non-rotary situation, wherein the fluorescence that the microparticle in the partial volume being irradiated with a laser is launched can be by detector
(15) this partial volume detected, is defined as detectable volume, and the method for estimating detectable volume is as follows: will launch
The fluorescence reference of fluorescence be fixed in sample cavity (6), and make detector (15) that can identify that laser beam is along the negative side z
To incident sample cavity (6), when sample cavity is fixed, the intensity of the fluorescence signal of detector (15) record is(formula one), wherein w0It is laser beam in the with a tight waist of the direction x
Diameter, z0For the beam waist diameter of laser beam in the z-direction, adjusts displacement platform II (5) and sample cavity (6) is translated in the x-direction, detect
Device (15) records the fluorescence signal of fluorescence reference, obtains w after being fitted using dimensional gaussian distribution to data0, adjust position
Moving stage II (5) translates sample cavity (6) in the z-direction, and detector (15) records the fluorescence signal of fluorescence reference, using two dimension
Gaussian Profile obtains z after being fitted to data0, it is fitted using formula a pair of the data obtained, estimates detectable volume, allusion quotation
Offset is 0.5 nanoliter；Detector (15) continuously records the fluorescence that the microparticle in a period of time T in detectable volume is launched
Intensity, obtains the signal data of some varying strengths, and computer obtains to visit in T time after handling the signal data
The quantity of the microparticle in volume is surveyed, and combines the revolving speed of sample cavity (6), the number of microparticle in unit liquid volume can be obtained
Amount.
Apparatus of the present invention have the sample cavity and stirring sheet of special designing, enable to the sample of different zones in sample cavity
Fast exchange is carried out, mechanical noise is significantly reduced, meets the needs of Ultrafast spectrum experiment, in addition, using rotation sample cavity
High-precision counting is carried out to microparticle in conjunction with the method for fluorescence detection, no sample loss is easy to operate.
Claims (1)
1. a kind of Ultrafast spectrum research device of microparticle, including displacement platform I (1), rotation motor (2), rotary shaft (3), magnet
(4), displacement platform II (5), sample cavity (6), stirring sheet (7), lens I (8), beam splitter (9), laser (10), optical filter (11),
Lens II (12), aperture (13), lens III (14), detector (15) and computer, xyz are three-dimensional coordinate system, rotate horse
It is fixed on displacement platform I (1) up to (2), the laser (10), beam splitter (9), lens I (8) and sample cavity (6) successively form
Input path, the sample cavity (6), lens I (8), beam splitter (9), optical filter (11), lens II (12), aperture (13), lens
III (14) and detector (15) successively form emitting light path, and laser (10) the adjustable continuous laser of launch wavelength and can swash
Light pulse, laser pulse include pumping pulse and direct impulse, and the pumping pulse and direct impulse have different power, frequency
Rate and duration, the time interval between direct impulse and pumping pulse can be adjusted,
It is characterized in that: magnet (4), by rotary shaft (3) connection rotation motor (2), rotary shaft (3) is parallel to z coordinate axis, it is displaced
Platform I (1) can be three-dimensional mobile, and rotation motor (2) is able to drive magnet (4) rotation, and sample cavity (6) is fixed on displacement platform II (5)
On, displacement platform II (5) can be three-dimensional mobile, and displacement platform II (5) is able to drive y direction center of the sample cavity (6) around sample cavity (6)
Line rotation, sample cavity (6) top half be rectangular body cavity, lower half portion be the cylinder barrel of axis in the y-direction, the top half
There is aperture plate between lower half portion, the rectangular body cavity is 30 millimeters a height of, 10 millimeters a length of, width is 2 millimeters, the cylinder barrel
A height of 20 millimeters, basal diameter be 10 millimeters, the axis collinear of the y direction center line of the rectangular body cavity and the cylinder barrel,
Stirring sheet (7) is wrapped up polytetrafluoroethylene (PTFE) and is made by having outside magnetic stainless steel, the diameter of stirring sheet (7) is 1.2 millimeters, long
Degree is 13 millimeters, and stirring sheet (7) is located at sample cavity (6) top half, and the aperture plate can prevent stirring sheet (7) from falling into cylinder
Bucket；The rotation motor (2), laser (10) and detector (15) difference cable connection computer, can be adjusted by computer
Save the time difference between the laser pulse that laser (10) issue and rotation motor (2) rotation angle；The size energy of aperture (13)
It is enough to adjust, the detection resolution of detector (15) can be adjusted according to various sizes of microparticle to be measured.
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| CN110395689A (en) * | 2019-06-24 | 2019-11-01 | 金华职业技术学院 | A kind of assemble method of microparticle |
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