TWI257705B - Device of detecting nanoparticle and method of the same - Google Patents

Abstract

A mass spectrometry system includes a first ion trap that selectively ejects charged particles based on their mass-to-charge ratios. A second ion trap receives the particles ejected from the first ion trap, causing the particles to gather near the center of the second ion trap. A laser beam is directed towards the particles in the second ion trap to induce fluorescence, which is detected by the photon detector. Particles are periodically dumped from the second ion trap. A mass spectrum of the charged particles can be obtained by comparing the photon count with the particle ejection characteristics of the first ion trap.

Description

1257705 玖, invention description: , * [Technical field to which the invention pertains] The present invention relates to a detection device, and more particularly to a nanoparticle detection device. [Prior Art] A mass spectrometer can be used to determine the type and amount of components that make up a solid, gaseous or liquid sample. Mass spectrometry uses the mass/charge ratio of ions to add 1 〇) to perform the work of separating and separating ions. The ionic charge represents the number of charges of the ions. The ion berry can be 1 unit (amU) or Dalcant (Dalt). 〇n, Da) to express. The quadrupole ion trap mass spectrometer (quadrupole ion trap mass QITMS) can be used to analyze the atomic, molecular and cluster ons of Berry. A typical quadrupole ion trap mass spectrometer has a ring electrode ( Ring electrode) and two cap electrodes (4) "叩electr〇d (8). Operationally, the force is applied to: the time_varying voltage between the ring electrode and the poles to cause the device to generate a The electric field that changes with time is limited; 卩 疋 疋 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 The light can be used to detect the presence of fluorescent molecules (such as dyed molecules) or particles that are marked by light molecules. For example, a laser beam directly illuminates ions in an ion trap to induce fluorescence (1). The optical detector is used to detect the number of photons emitted from the ions. [Invention] The purpose of the present invention is to provide a way to detect the nanoparticles of the nanoparticles. Step: periodically ejecting the particles from a first ion trap according to the mass/charge ratio of the charged particles; a second ion trap is received from the first ion trap and throwing 1257705 particles:::: The invention directly provides a method for detecting nano particles, comprising the following steps: :: periodically throwing charged particles in a diion trap; The second ion trap receives the particles ejected by the dice trap; and detects the fluorescence induced by the second ion. The above and other methods of the present invention may include one or more of the following features. An ionic body throws charged particles according to the mass/charge ratio of the particles. The method comprises directly irradiating a laser light source to a solid, liquid or colloid sample to desorb and ionize the particles in the sample to produce the first - an ion trap. m. The method comprises using an electrospray ionization (electr〇spray i〇mzat_) technique to generate charged particles for supply to the first ion trap. The method comprises direct illumination - the source of the rake originates from The second ion is tempted The guiding particles generate fluorescence. ^ The method comprises periodically removing all of the particles in the second ion. The method of capturing the light source comprises using a photomultiplier tube (10) to detect the number of photons thrown from the particles. The photomultiplier tube is cooled to a temperature lower than zero degrees Celsius. 5 Haiguang Xunyi calculates the number of photons in a idle period (gatepen〇d) and generates a count value. All particles are in the interval period. (dweiitime), removed from the second ionic taste. The method comprises labeling charged particles with fluorescent dye molecules. The smectic molecular marker charged particle system comprises more than one type of fluorescent dyed molecule Mark charged particles. 7 1257705 The method comprises producing a mass spectrum of particles labeled with a specific type of dyed molecule. The method includes controlling an alternating voltage signal applied to the second ion trap such that the particles received by the second ion trap have a damping time of less than 200 milliseconds. The method includes applying an alternating voltage signal to the second ionic body to cause an oscillating electric field to confine the particles to the second subwell. The method includes periodically turning off the alternating current, .... all the particles in the ion trap, and applying the DC voltage to clear the particles counted in the gate & gate time. The particle system is a fluorescent dyed particle. The particle system contains fluorescent dye molecules. The particle system contains biomolecules. The particles received by the second ion trap have a mass/charge ratio higher than 106. The invention further provides a device for detecting nano particles, comprising: _: a card receiving particle sub periodically from the mass/charge ratio; - a second ion card received from the first detachment Particle = Drag Detector: Fluorescence induced by particles detected from the second ion. And - other methods, which may include the following - or several features. ,,~2 l is called "the tiger generator, generates a voltage 随 that changes with time, * is applied to the second ion electromagnetic field, • throws from the first - ion card:::7:: card f- After the shocking castle is decelerated, the scorpion; the child enters the second ion trap in the middle of the second disengsing position, that is, the first ion trap system includes a quadrupole of the first ion _ is determined to periodically throw two ions of 1,257,705 meters. . . The first ion trap is set to periodically throw, .. M 'he is higher than 106 particles 5 Hai second ion trap system includes one four The polar ion trap includes a fluorescent detector. The device includes a laser generator that generates a laser beam of the medium particle. The second ion trap device includes a first signal generating device. And generating a trap to generate a "seismic" signal in the first ion. The first child of the first child, which changes with time, is the first frequency of the first-electric signal frequency during a measurement cycle. The first frequency to one: the change of the morphological relationship between the morphological and the mass of the particle The frequency is periodically thrown such that the first ion the first signal generator scans the variability over time' such that the frequency change is linear with time. The voltage signal frequency of the first signal generator scans over time: the two frequency changes have a nonlinear relationship with the time, (4)::: The number of particles thrown in the "Hai Di-Ion card, its mass/charge ratio and During the time = period, the device includes a second signal generator to generate a linear relationship. The second ion device is generated in the second ionic morphine. Including - providing an ion supply source for the particle. The σ海 ion supply source includes a particle-containing substrate. The first ion trap is included from the first ion trap, and the following steps are performed: the ion trap is thrown to electrify the first ion trap system and the second Ion trap bonding, the thrown particles are captured by the second ion trap. The present invention further provides a method for detecting nanoparticle using a second ion trap to reduce the periodicity of the velocity from a first - 1257705 particle; A fluorescent light source is induced by a laser light source and is emitted from a charged god. The present invention further provides a light, eye-catching method for measuring the negative particles of the clothing, including the following steps: The flow of thunder ' 爿 爿 , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , Throwing charged particles; applying a 第-丄^ ' ^ 4α A ^ 弟一父, machine 黾 信号 于 于 苐 苐 苐 苐 苐 苐 苐 苐 苐 — — — — — — — — — — — — — — — — — — — — — The predetermined amount of the first-to-AC voltage signal describes the frequency of the second alternating voltage signal such that the particles receiving the ion-charge are confined to the second ionic.

Other methods of the invention may include one or more of the following features. Knowing the second AC voltage, the tiger frequency keeps the second ion card attached (qz) unchanged, which is related to the second ion_receiving particles. The ... wheat number (6) is proportional to the amplitude of the second alternating voltage signal and inversely proportional to the frequency of the first alternating voltage signal. The method includes periodically reducing the amplitude of the second alternating voltage signal to eject particles in the second ion trap.

The present invention further provides a method for detecting nanoparticle, comprising the steps of: controlling a signal in a first ion trap such that the first ion trap periodically ejects ions according to an ion mass/charge ratio; a second ion is collected from the ions thrown by the first ion trap; detecting fluorescence emitted by the ions in the second ion trap to generate a beech indicator; and connecting the detection signal and the control signal to A mass spectrum of ions in the first ion trap is determined. The above and other methods of the invention may include one or more of the following features. The method includes directly irradiating a substrate with a laser beam, and desorbing the nano biological sample to generate ions in the first ion trap. No, the method includes directly illuminating a source of laser light in the second ion trap to induce fluorescence. 1257705 The method includes adjusting a parameter of the second ion trap to conform to a parameter of the first ion trap, causing the second ion trap to capture ions emanating from the first ion trap for at least one specific time period, and detecting from Fluorescence emitted by ions in the second ion trap. The method includes adjusting a parameter of the second ion trap to conform to a parameter of the first ion trap such that the second ion trap captures ions having a specific range of mass/charge ratios from the first ion trap. The method includes periodically ejecting ions from the second ion trap. The method includes generating a fluorescent signal of ion emission from the second ion trap, the ions being subsequently purged from the second ion trap.

The invention further provides a method for detecting nano particles, comprising the following steps: - the second ion card receives ions periodically thrown from the -first ion according to the _f/charge ratio, and is induced by a laser Ion emitting fluorescence; and adjusting parameters of the second ion trap to conform to parameters of the first ion trap, causing ions to be confined to the second ion for at least a period of time, and detecting ions from the second ion The laser-induced fluorescence of the emission (lase, m, and other methods of the invention may include the following - or several features. The ion system is a fluorescent substance or a substance containing a fluorescent molecule. A device for detecting nanoparticles, comprising: a second sub-solution 'receiving basis-quality/charge ratio periodically from the first-ionic morphine throwing: a sub-a laser generating ϋ, generating - directly illuminating the first The second ion::::beam; the photodetector 'detects the egg from the second ionic morphine, and the -circuit, produces - the pressure applied to the second ionic, causing the second ion (4) Selecting time to clear the particles, the The clearing of the sub-work to the interval-specific time period 'to enable the photodetector to probe the above and other methods of the present invention may include the following - or a plurality of beta-diaphragm-ion cards comprising a quadrupole having two cap electrodes Ion card. 1257705 ... This mourning includes a 彳5 tiger generator, generating a control voltage applied to the first slave: the second control voltage having a scan-first frequency to a frequency of a brother-frequency In order to make the above-mentioned objects, features and advantages of the present invention more apparent and easy to understand, the following description of the preferred embodiment and the accompanying drawings are as follows: Method] The embodiment of the dual ion trap mass spectrometer is described in Figures 1 and 2. A double ionic glycan mass spectrometry system 100 includes a first ion two of the particles according to the mass/charge ratio of (4). The diion _ 1G4' collects and slows down the velocity of the ejected particles, indicating that the ϊ = is near the center of the second ionic body; a laser beam 150 is directly in the tube; t is in the second 〇 4... Fluorescent, and borrow - photomultiplier clothes, 'f The electro-particles are periodically cleared from the second divergence 4 at predetermined time intervals, and the fluorescence intensity detected at each gate time is generally proportional to the number of particles of the qualitative/charge ratio. Tube 1〇6期二:=Γ=Γ and the first (four) 102 in the measurement of the week's particles = after conversion can get a second ion _ just received the first card 102 and the second ion card 104 series Set to always: 2: two-ion... is, for example, a hyperboloid quadrupole ion two=(four) pen electrode 108, a first cap electrode 110 and a second cap electrode 112, and the ring V electrode (7) 8 has two positive The relative position of the hole 120盥122. Double: The morphine mass spectrometer 1〇〇 can be used to measure particles with a wide range of sizes, ^, particles at 10 nm, particles with a mass greater than 106 dow, to 12 4 1257705 Particles with a mass/charge ratio greater than Η)6. The charged particles themselves may be glory substances or substances marked by fluorescent dye molecules. In the example, a matrix-assisted laser desorption ionization-rix-assisted laser des〇rpt job (10) (10), MALm) technique is used to generate charged particles in the ion block 102. A sample 18 is placed on the surface of a non-mineral steel sample rack 126. The sample can be a matrix containing the particles to be analyzed. A laser beam 121 is incident on the hole 120 and passes through the hole 122 after passing through the first ion _ 1 〇 2 to cause desorption and ionization of the particles. The first pin drive signal M2 of each frequency f丨 and a voltage amplitude Li is applied to the ring electrode 1〇8, and the cap electrodes ιι〇 and ιΐ2 are grounded to limit the charged particles to the first ion card 102. The first taste drive signal 142 is generated by a # generator 146 and amplified by a power amplifier, 144. The frequency range scanned by the number generator 146 depends on the mass/charge ratio of the particles to be analyzed. A computer 150 transmits a trigger signal 152 to trigger the signal generator 146 to initiate a frequency sweep, which is specific when the frequency f is swept from, for example, a higher frequency to a lower frequency (e.g., 60 kHz to 500 Hz). The shift of the mass/charge ratio particles will begin to become unstable and the particles will be ejected from the (five) 124 of the first ion electrode. If the card parameter of the first ion $1〇2 and the voltage amplitude ^ 疋 value, the f / charge ratio of the ion ejected from the first ion It and the frequency [present a function similar to the first ion clock 102, second The ion 〇1〇4 may be, for example, a bipolar ionic morphine including a ring electrode H4, a - cap electrode 116, a cap electrode 118', and a first cap electrode ιΐ6 provided with an ion that allows the first ion i to enter the second Hole 128 of ionic brown 104. A second well drive signal 176 comprising a frequency f2 voltage amplitude vac, 2 is applied to the ring electrode. The cap electrodes 1 i 6, 118 are then grounded. The first well drive signal 176 is generated by a signal generator 178 and amplified by a 13 1257705 power amplifier 180. The second well drive is "": 178 can be controlled by the computer 150." Transparency and selection of voltage amplitude An ion ejected by the H-ion card 102 can be confined to the second ionic film 104. Within the lift (4) parameters, said, the number of hours (6), can be described to describe the motion of a gamma electric particle within the ion. Sound Ding (Formula 1) — 〇n/z)r〇2Q2 Pressing wipes:,,: '2 is the particle charge' is the radius of the electric cover electrode of the spell drive signal, driving the frequency of the money. Tian H〇. When helping, the ions become unstable and will be thrown from the ionics. The molecule ^ just captures the charged particles emitted by the outside (can be atomic or 104 ^ >;::7: Γ02 particles are captured by the second ion $104, dragged in the second ion card (10) =, The number of the card is different from the estimated card parameter qz2 of the particle in the whistle. When the first, second::: units (for example, etc.) are the same, the relationship between (10) and ! is deleted. Can be described as follows·· 4,2

Qz ~ Reject,] ΩιΨογ, Ι ω7κ7λ (Form 2) 八1 2,FacjfGc,2) are the well drive frequency and voltage of the first ion trap 1〇2[笫-hetero well 1〇4), Because of the (different-displacement drive frequency and voltage, the #^ 舁二二离子井's 102 throws a large number of f: sub-two: '2 value is less than 〇·9〇δ, for this reason, from the first-ion fat Large particles can enter and be captured in the second ionic body 104. Male, and the H drive signal scans the _ frequency range when the seven values will change. The method of changing the value of ", ., qz, 2 in the scan range lies in While scanning the first-ion fat frequency, it also scans the second from y-be and half to keep the value of qz, 2 at 14 1257705 ^ by computer 1 5 () synchronous control of the first and second ion trap sweep For example, in the case where k is equal to L, 2, synchronous scanning, and the scanning frequency is 33, qz can be obtained during scanning, and the value of 2 is 0.1. By circulating the charged particles in the diionic odor 104, A laser illuminating 50 diion 152 directly illuminates the 15th 〇 excitation with a 胄 density particle distribution / due to the laser (or fluorophore on the particle) is laser The beam shovel V is ready to pass through the hole 130 of the end cap electrode H8, and continues to count by the number of _ male juices of 164 yuan, and the count value of the photon counter 164 is transmitted to the computer 1 5 于 in the following step. The space 160 with the particle:::, ion spliced 1 〇 4 is filled with buffer gas to slow the incident laser beam (9), day: V, 4 region, so that the focus is stagnant in the second ion into the second ion...04 Low resistance slow time can be increased as /; or ratio ^ is called "resistance time", drop (resolut1〇n) 〇... scoop noise ratio (four) naI-t〇-n〇lse) and resolution second ion The well 1〇4 will be periodically thrown out. The count value obtained by the hunting f sub counter 164 is roughly the same as the second deviation.

The number of two sub-numbers is proportional to. 4 Clear the particles, temporarily turn off the first driving signal 17 6 ' and apply the DC clearing to do m pi j · JU Bu, Pmg Slgnal) 181 in the end cap: '¥ f electric particle 攸 second ion _ 104 hole 130 clear, straight "except for the polarity change of the signal is based on the polarity of the charged particles and the stator ^ charge, then the DC clear signal is negative power, and vice versa (4), j fruit particles are not periodically thrown, later 如 ' as early as The particles entering the spell 104 will emit photons for 15 1257705, causing repeated counting. The photon counter 164 is counted in the photon/^ (_ Ume), which is detected by the 卩 日 日 日 日 日 日 日 日Zero back to zero. By: the number is then followed by an interval of time 1"-), and the specific time (determined by the count value) occurs simultaneously in the light ^ U. The connection is a certain force 々 * the intensity of the fluorescence and the frequency of the well drive signal can be determined = : The difference in the quality of the medium, the particle characteristics of the charge ratio, the fourth... The system reveals the mass spectrum obtained by the above method. The time limit of the closed time and the interval time of the spectrum of a special type f particle is determined by (4) ^: The slowing time of the particles in the wide 1〇4 is determined by scanning the first-dream light (four) rate to throw the particles to be analyzed, and then the counting slows down; the man falls. The particle is confined by the electric field to the center of the second ion trap 104 = strong:; first there is a high peak /. When most of the particles are concentrated near the center of (4) 4, the particle insects are again and the other X molecules - particle collisions (for example In the second ion Shaanxi, the influence of 'charged space =! = collision between molecules', in addition, the slowing time is also affected by the local distribution, that is, due to the charge repulsion between charged particles, making it near the center of less m The time 'will be longer than the limit T. The time of the child is long. Reducing the de-drive voltage will cause a slowdown, 曰ϋ 'because the solution | zone dynamic voltage drops a little longer time stealing:: the particle's motion is expanded' so it has to be chosen. Near the center. Since the short slowdown time is better, choose the second formula = give the second ion taste 104 a larger brown drive voltage, but if 'two one:, 4'2 can't be too high' would otherwise Qz, the value of 2 is higher than 0.908, and the particles in the sub-iQ4 are unstable. Thus, the voltage amplitude and frequency of the first and second departures k唬 must be chosen such that the value of qz, 2 remains below the value of 〇 9〇8 16 1257705. For more information on the stomach, see Figure 3, Figure 224 shows the waveform of the trigger signal 152, and the 226 shows the waveform of the frequency U Tiger 148. The target 172 shows the waveform of the DC clear signal, "including the periodic voltage pulse (penodlc v〇ltagepulses) 232, the picture is the second:: the on signal of the on-off signal of the dynamic signal 176, the head 23" shows the laser from the laser Inducing the counting of the fluorescent photons, wherein the high point "238" indicates that the photon counter 164 is counted from the fluorescence of the second ion_ganger particles, and the "low point, (10) indicates that the meter has been reset to zero. Without counting, please display the count value of the photon counter 164, thereby constructing a mass spectrum of a charged particle. At time U, the trigger signal 152 is sent to trigger the generation of the scan frequency signal. The DC clear signal is at the low point 220 and the second brown drive signal 174 is turned on = 22. The particles accumulate in the second ionic membrane (10) in this environment, and the photons from the laser-induced fluorescence are counted 222 from the photon juice 164. At daytime t2 (ttl~t2 is the time between photon counters 164), the second morph/driver 176 is turned off, and the dc clear signal 172 is turned on, which causes the charged particles to be cleared from :::#104. The positive or negative of the DC clear signal varies depending on the polarity of the charged particles. In addition to the wide t3 (t2~t3 is the interval time of the photon counter 164), the DC 2 is turned off and the second fat driving signal 176' is turned on to cause the charged particles to open in the young ion trap 104. The gate time of 164), the second well drive causes the charged particles to start from the first time t4 (t3~t4 is the photon counter signal off, and the DC clear signal is turned on, the diion trap 104 is cleared. At time t1 to The t2 phase has a mass/charge ratio determined by the charged particle H8 accumulated in the second ion trap 丨〇4 between the count values generated by the scanning frequency signal time t2. The photon counter 164 is substantially coincident with the time t1 and The average of t2 is 17 1257705. The frequency is a functional charge/mass ratio of charged particles. At time t3, the charged particles accumulated in the second ion _ 1〇4 before time t1 to t2 are almost cleared, therefore, photons The count value produced by counter 164 at t4 generally corresponds to the number of charged particles having a charge/mass ratio as a function of time t3 and the average frequency of the jitter. According to the above method, the first well drive signal 142 is scanned to - Low frequency 23. In the period, just the first ion two: 2; after the electric puller, the photon number of the charged particle can be constructed by using the photon counter 164 at the end of each gate time. 178. Implementation of the dual ion trap mass spectrometry system In one embodiment, the first spell 1 〇 2 and the second morphine 1 () 4 are RM J〇rdan = pany, paul Gra of Grass Vauey, Calif〇rnia The first and second ions: ... 〇 mm, z. is 7.07 mm 'where r is the + 环形 of the ring electrode i 〇 8 , Z 〇 is half the center distance of the two cap electrodes 110 , 112 . The twist is 2 mm , with a radius of 3 mm mm ring

+ on the core electrode, as the introduction of the charged particles and the MALDI:! beam 121, the injection/exit of the probe laser beam 150, and the radius of the hole on the first ion (four) 2 of 31 In millimeters, the radius of the hole of the cap electrode and the ring electrode of the t-meter 104 is respectively and the bottom of the mechanical pump (not shown) is used to exclude the pressure in the vacuum chamber 132: it is lowered to a low At a base pressure of 1 mTorr, another helium gas is introduced into the chamber with a force of 50 mTorr (4) ♦ gamma epressure) (1) The bronze laser beam 121 is a 5 mJ/pulse (energy), 355 Nai The pulsed laser 'produces from - triple frequency (- ency seven... (10) toe from g 18 1257705 laser 138 (model SureliteTM, from Continuum, Santa Clara, California). Laser beam 121 is borrowed by a focal length of 0.5 The meter-long lens 140 is focused to form a concentrating spot having a radius of substantially 1 mm on the sample 180. The signal generator 146 is a model DS345 Function & Abritrary Waveform Generator (from Standard Research System, Sunnyvale, California), and borrows a computer 150 Data acquisition program Acquisition program, Labview, from National Instruments, Austin, Texas) The laser beam 134 has an operating energy of 400 to 600 mW and a wavelength of 488 nm. It is generated from an argon ion laser 154 (model Innova 90C, from Coherent Inc., Santa Clara, Calif.) The laser beam 150 is focused by a lens 156 having a focal length of 1 meter, continuing through a light baffle 158 and a hole 152 of the ring electrode 114 of the second ion trap 104 to form a A generally 200 micron spot is concentrated at the center of the well. Fluorescence emitted from the charged particles is focused by a lens system 136 having an aperture value (F numbe 〇 3 and a focal length of 38 mm. The photomultiplier tube 106 is a thermoelectrically cooled ( Thermoelectrically cooled) photomultiplier tube (model R943-02, from Hamamatsu Corporation, Bridgewater, New

Jersey). The photon counter 164 is model SR400 (from Stanford Research System) ° The particle system to be analyzed is polystyrene beads (FluoSpheres, from Molecular Probes, Eugene, 〇regon) suspended in water and marked with yellow-green fluorescence. According to a measurement result, the size of the polystyrene ball is 27±4 nm. Another measurement result, the size of the polystyrene sphere is 1丨〇±8 nm, of which 27 nm and 11 nm. The spheres contain the number of fluorescent dye molecules of 18 〇 and 74 分别, respectively. The absorption wavelength of these polystyrene spheres is 490 nm, and the emission wavelength is 515 nm. The quantum yield is 30%. To prepare a sample of MALDI, the suspended particles were diluted with deionized water to give a concentration of 19 i 1257705 ίο particles per cubic centimeter. An equal volume of sample, matrix 盥 ( acet〇ne, 5 solution is mixed together and placed on the surface of sample holder 126, sample holder 126 electrode (10) and embedded in the hole of the ring electrode above the first ion plate. ^Diion _ space is filled with helium at a pressure of 5G mTorr

Buffer gas, brother I? iUi- 1 AO maldi generated charged particle nitrogen buffer gas assisted capture by the body to assist the η:, the helium buffer gas in the brother-ion trap 104 assists in capturing the trap by the first ion trap 102 Out of the particles. ^ ^ ^ ^ ^ 〇2 - « ° -^^^ctlon plate) Cheats to suppress the pressure to increase detection efficiency. Nightmare 2:: Size of 27 nm of polyethylene particles mass spectrometry, first ion quality:; not ... 0 ° Hertz card driving frequency (%) operating in - axial pad pattern (axlalmass_selectlve ^ In the presence of buffer gas, the m-gate production method avoids the arc discharge phenomenon between high voltage and a Hongji. It is known that the 27 nm particles in the 27 nm polyethylene particles have a slowing time. The L:V:" diionic morphine 200 reads from 6 kHz to 〇 'year (Απ) is a photomultiplier tube 1 〇 6 detected fluorescence 1 to throw 27 nm particles. In Figure 4, Ml 5 〇 托 ,, rise day (four) 4 〇. 2 seconds. In the ^ inch '(four) gas pressure is 60 volts, knot seven Figure 4β, the voltage amplitude Γ, according to the above results, the force is 5 〇 mTorr, then rise Time 4 (J seconds. Polyethylene, count between children: = photon between the materials, selected as 27 nm clearing time is 2 milliseconds, DC clearing signal is · ι volts, above 20 1257705 is enough Let the charged particle 5A show that the two sub-wells 104 are cleared and avoid particle accumulation. Mass spectrometer 200, which is about two square meters & The single-scan of the ball is from 2*1 〇6 to 6.5 million dolphins, and the signal/magnitude distribution of the Fandi 5β image shows a signal contour of 2〇2. Mass spectrometry 202 accumulating ten single-scan results, the mass spectrometry is mass/charge ratio concentration, and the mass spectrometry is smooth. The most important feature of the mass spectrometer 202 is based on the previous 1 micron syllabus 2·5 1 〇, mainly For the contribution of singly charged particles, 0.908) is corrected. In the case of NMR, the mass spectrometer uses the throw point of 〇·95. In the 5A and 5B diagrams, the second well drive signal 176 has a fixed frequency (Ω%). He Xuan’s 5 C picture is also shown; in the spectrum, the _ cumulative ten-single-scan results of mass spectrometry, because the quality number 142 is the same as:: moving 'number 176 frequency, the female D is fixed as (u. Bibi The human kilohertz, and the card parameter detection range is shifted to, the handle knows the shell, the 曰204 special (for example, the scan changes the second off/ho ratio & field', which means the dynamic capture condition is fixed first. = the frequency of the signal) compared to the static capture condition (eg ion 1〇4_. 频率 said frequency) there are more double-charged particles displayed by the second brother 6A map _ borrow scan 1 〇 kHz to Drive signal frequency (Ω1/) and 婶...a He Hyun's brother-in-the-pot plant A and get a single-scan mass spectrum 210 of 110 nm particles, ...:: Volt. The drive signal makes the -ion card 1 2 is thrown in the range of Nayong 6 to (10) * 106. __ The average quality of the viewing beads is ..., Wan Daoer five " 〇 million Dao Tun, the mass distribution is 350 ~ 543 hundred 禹 尔 吞, The difference between the dimensions is shown as ^8 奈. This shows the particles carrying 1 to 6 charges 21 1257705 = the mass/charge ratio range is (350~543)*106 to (58~91)*1〇6 between. Figure 6B shows a mass spectrum 212 that accumulates ten single-shot results, the characteristics of which are derived from multi-charged particles. Comparing mass spectrometry (1) with mass spectrometry 2〇4 (figure %), it was found that the number of charges carried by the U-nanoparticles was approximately twice that of the 27 nm particles. Here, the particles which are produced by MALDI and are lag behind the first ionic morphine 1〇2 are evaluated. Generally, i-o% will enter the second ionic morphine 4 according to the axial mass selection throwing mode, because (1) the first ion trap The two cap electrodes of 1〇2 throw an equal amount of particles. (7) During the transfer of the first ion 102 to the second ion card 1 () 4, a part of the thrown particles may be lost. (3) Due to the phase difference mating, some of the particles cannot be captured by the second ion card 104. Therefore, for example, if there are _ charged particles in the first ionic morphine 1 〇 2, the laser-induced fluorescence method will be used to scan the entire frequency range after the second ion plate H) 4 will be # 1 〇 (M solid particles are It is detected that since the particles measured in the 5A~6B map are particles with little difference in mass and charge number, for example, the peak with better separation effect (such as 214, 216) will appear in the low particle density, such as the 6A map. As shown in Fig. 6A, the features of the sharper tip and better separation effect (such as μ, which may be due to the number of H0 nanoparticle particles per particle containing the total number of fluorescent dyes) can be easily detected. It is also possible that particles are labeled with iq or fewer fluorescent molecules and detected by the above method. Since particles of any size can be labeled by dye molecules, the 遂 dual ion card mass spectrometry system can have a broad mass spectrum. Scope of analysis. The dual quadrupole ion mass spectrometry system 100 can perform mass analysis of biological macromolecules or biological particles 'for example' system 1 〇〇 for detecting leaks _iarpr-s company produces fluorescently labeled IgG (goat antl_m ( ) use antib〇dy). An average of 6.2 Alexa Fluor 488 luminescence molecules (mass 643 Dolby) were labeled with a total mass of (10) thousand dolphins, and an Alexa dye of 22 1257705. The maximum absorption and emission wavelengths were 497. Nano and 518 Nai. The laser used in the 27 nm polystyrene beads is the same as the light collection system for measuring the mass spectrum of the labeled IgG molecules. When measuring IgG, the gate time is chosen to be 2 〇 milliseconds. The DC clear signal applied to the outer electrode of the second ion trap 1〇4 is 2 volts. A single frequency scan is obtained in 5 seconds of data point data in n seconds. Sinapimc acid is used as laser desorption/ionization The matrix of the 7/7 image shows a mass spectrum of a camping light IgG molecule obtained from the first ion trap driving frequency of the 30 kHz to 5 kHz broom, and the temperature is obtained by \α", 1 is 200 volts. And the second ion trap 104 operates in a dynamic capture mode (for example, the frequency of the first-ion w drive signal 176 is simultaneously scanned with the frequency of the first (4). 1 port says a piece... a quality cast (maSS re- ) (1=5) can be accumulated by ten scans, and the order ratio is 1·5* 1〇 The data of the charge 1gG molecule is obtained with a noise ratio greater than 10. The double hybrid card spectroscopy system can be used to analyze the biomolecules of biological giant particles = because the dyeing markers are often used in life sciences. The above method is quite practical. Previous mass spectrometry, Bicolor:: (4) by means of light to detect the number of dye molecules attached to the biological particles to add the mass of the dyed molecules on the sub-), and the system ~ and the knife does not need to carry multiple charges The advantages that can be detected. 隹U本明明 has been disclosed in the preferred embodiment as above, shovel & hair: Anyone who is familiar with the art, can not be changed from this = two =: And the retouching, therefore, the companion of the present invention = sub-neutral scope, please define the scope of the patent defined by the scope of the patent A _ ', ° and 干 砚 砚 砚 。 。 。. Other embodiments are described below in terms of patent scope. 23 Ϊ257705 The handyman in the first ion trap 1〇2 + the mass male Mr can be electrically sprayed with ionization. 5th to 7th

The Beikou daily map is a poem obtained by linearly scanning the first well driving frequency under a low voltage (2〇0 you 4 士, ^ J 叮, then P (00 ”), which can avoid the pressure (5〇喜k" If you have a bad corpse, you can get a linear scan of the frequency at which the electrode arcs to the computer 150. The 质谱 」 仔 在 在 在 在 贝 贝 贝 贝 贝 贝 贝 贝 贝Morphology 1 〇 2 is temporary, g 炫 π 竹 竹, technology μ, carved in the unstable mode of the trap 102 in Berry remotely ^ she twisted across the two cap electrodes of the auxiliary AC voltage scanning well drive voltage Amplitude. - The different functional fauna is set between the first and the second ion plate. - The helium pulse is applied to the first ion _ in order to store the first ion block 7: the particle 'and the second ion. Maintaining a steady flow of helium buffer gas is the purpose of slowing down the particles. The mass spectrometer 100 uses a % optical lens system to increase the light collection efficiency to increase the sensitivity of the detection, and the sensitivity can be further extended by using a larger opening. The capture device is boosted by reducing the degree of scattering of the background laser light. A light laser diode (blue d1〇de laser) or a high-energy LED (wavelength of 473 nm) can replace argon ion laser as a light source to reduce cost. Staining calibration technology for specific products (sample- The specific dye-labeling technique can be used to multiply the nanoparticles in different samples by laser-assisted multicolor fluorescence spectroscopy. 24 1257705 [Simplified illustration] Figure 1 and Figure 2 According to one embodiment of the present invention, a dual ion trap mass spectrometer is not intended. Fig. 3 is a graph showing changes of different signals over time according to an embodiment of the present invention. Figs. 4A to 7 are diagrams according to the present invention. One embodiment, the mass spectrum of the particles. [Description of symbols] 100~ dual ion trap mass spectrometry system; 102~ first ion trap; 1〇4~ second ion trap; 106~photomultiplier tube; 108, 114~ ring electrode 110, 112, 116, 118~ cap electrode; 120, 122, 124, 128, 130, 152~ hole; 121, 134, 150~ laser beam; 12 6~ sample holder, 132~ chamber; 136, 140 , 156~ lens; 138~Nd:YAG laser; 142~1st ion trap drive signal; 144, 180~ power amplifier; 146, 178~ signal generator; 148~ scan frequency signal; 150~ computer; 152~ trigger signal; 154~ argon ion laser; 25 1257705 158~ Light blocking plate; 160~ internal space of the second ion trap; 162~ fast preamplifier; 164~photon counter; 170~3; 172~DC clear signal waveform pattern; 176~second ion trap driving signal; 178~photon counter count value map; 180~sample; 18 1~DC clear signal; 200~27 nanometer polystyrene sphere single scan mass spectrum; 202~ cumulative ten times 200 mass spectrum; 204~ cumulative ten times 200 Mass spectrometry (detection of low mass/charge ratio region); 210-110 nm polystyrene sphere single-shot mass spectrometry; 212~ cumulative ten times 210 mass spectrum; 214, 216~ peak; 220~DC clear signal low point; 222~Second ion trap drive signal is turned on; 224~ trigger signal waveform pattern; 226~ scan frequency waveform pattern; 228~ scan frequency signal high frequency; 230~ scan frequency signal low frequency; 232~cycle power Pressure pulse; 234~second ion trap drive signal on-off diagram; 236~laser induced fluorescence photon count diagram; 238~laser induced fluorescence photon count high point; 240~laser induced fluorescence Photon count low; 26 1257705

Claims (1)

Amendment date: 94.7.21 1257-year-old 25,852 patent application scope revision, patent application scope: L method for measuring nano-particles, including the following steps: Zhou Shengsheng from a first ion trap (ion trap) The charged particles are thrown; the second ion card receives the charged particles thrown from the first ion fat; and the fluorescent light induced by the charged particles is detected from the second ion. The method of detecting nanoparticle according to the above aspect of the invention, wherein the first ion card throws the particle according to a mass/charge ratio of the charged particle. & 3. The method for detecting nanoparticles as described in the application of the patent specification, further comprising directly irradiating a laser light source to a sample of a solid, liquid or colloid to desorb and ionize the sample The particles 'produce the charged particles in the first ion trap. 4. The method of detecting nanoparticle according to the above-mentioned claim, further comprising using an electrospray ionization ionization (orietn) spray iQnizat_) technique to generate the charged particle and provide the charged particle to the first ion trap . 5. The method for detecting nanoparticles according to claim 1, further comprising utilizing photoionization (phGtG_iGnizati()n) technology to generate the charged particles and providing the charged particles in the first ion trap . 6. The method of detecting nanoparticle according to claim 1, further comprising directly irradiating the laser light source with the particle + in the second ion to induce fluorescence of the σHai particles. . 7. The method of detecting nanoparticle according to claim 1, further comprising periodically removing at least a portion of the particles in the second ion trap. 8. The method of detecting nanoparticle according to claim 1, wherein the step of measuring the light source comprises using a photomultiplier tube (ph〇t_ltipHer 0578-A20102TWF2(N1); 07A-920505; david 28 1257705 tube) The number of photons thrown from the particle by a factor of five. 9. If you apply for the method of exploring the greens in the eighth item of the full-time, the photomultiplier tube in the basin is cooled to a temperature lower than zero degrees Celsius. The method of detecting nanoparticles as described in claim 8, wherein the photomultiplier tube calculates the number of photons in a gate period and generates two count values. 11. The method of detecting nanoparticle according to claim 10, wherein at least a portion of the particles in the second ion trap are removed during the interval. 12. The method of detecting nanoparticle according to claim </RTI> wherein the method further comprises labeling the charged particle with a fluorescent dye molecule. 13. The method of detecting nanoparticle as described in claim 12, wherein the charging of the charged particles is performed by more than one fluorescent dyeing molecule. 14. The method of detecting nanoparticle as described in claim 12, further comprising generating a mass spectrum of the particle labeled with a particular species of dyed molecule. 15. The method of detecting nanoparticle according to claim 2, further comprising controlling a voltage signal applied to the second ion trap to enable the second ion trap to receive the particle The damping time is less than 200 milliseconds. The method of detecting nanoparticle according to the above application, further comprising applying an alternating voltage signal to the second ion trap to generate an oscillating electromagnetic field to limit the particle to the second In the ion trap. 17. The method of detecting nanoparticle of claim 16, further comprising periodically turning off the alternating voltage signal to remove all particles in the second ion card. 18. The method of detecting nanoparticle according to claim 1, wherein the particle is a fluorescent particle in 0578-A20102TWF2 (N1); 07A-920505; david 29 1257705. The method for detecting nanoparticle according to Item 1, wherein the ruthenium particle system comprises a fluorescent dye molecule. 20. The particle system comprises a biomolecule as set forth in Patent Application No. 1. 21. The method for detecting nanoparticles as described in paragraph 106 of the first-half ion card of the first application of the patent scope, the method for detecting nano particles, wherein a part of the particles, The mass/charge ratio is higher than 22. A device for detecting nano particles, including a flute-ion plate, receiving particles and periodically ejecting the particles from the first ion trap according to their f/charge ratio a second ion solution, the particle received from the first ion, and a detector 'detecting the light source emitted by the particle from the second ion (four). Further, the apparatus for detecting nano particles described in claim 22 of the patent application method generates a voltage signal which changes with time when the whistle is applied, and generates a shock electromagnetic field when the applied ion (four)' The self-ion "extracts the particle, and the second ion (four) is decelerated when entering the person. = The device for detecting nanoparticles as described in the scope of Patent Application 帛22, &quot;苐-ion solution system includes a quadrupole ion trap. = For example, please refer to the device for detecting nano particles in the 22nd syllabus of the patent, and the I-ion ion-series system is set to periodically throw the size between 1G and 100nm ^ as described in item 22 of the patent scope. The apparatus for detecting nano particles, wherein the first ion solution system is set to periodically eject a particle having a mass/charge ratio higher than that of the crucible. 27. The device for detecting nano particles as described in claim 22 of the patent application, 578~A2〇1〇2TWF2(N1); 07A-920505; david 30 1257705 wherein the second ion trap also includes a quadrupole ion trap. 28. The apparatus for detecting nanoparticles of claim 22, wherein the detector comprises a fluorescent detector. The device for detecting nanoparticle according to claim 22, wherein the particle is a fluorescent particle. The device for detecting nanoparticle according to claim 22, wherein the particle system comprises a fluorescent dye molecule. 31. The apparatus for detecting nanoparticles as described in claim 22, further comprising generating a laser beam directly incident on the second ion trap. For example, the device for detecting nanoparticle+ described in the patent application 帛22 $, including the first signal generator, generates a time applied to the first ionic body to generate the oscillating electromagnetic field. Sinusoidal function letter! 3: The device for detecting nano particles as described in the Shen Qing patent range of $32, wherein the β hai-signal generator scans the frequency of the first voltage signal at any time during the measurement cycle - the first The frequency to - the second frequency fat is thrown off according to the mass/charge ratio of the particle. Ion=If you want to use the device for detecting nano particles as described in item 33, '5, the 5 tiger generator scans the first voltage _ 赛@ rate, which makes the frequency change linear with time. . The device for detecting nanoparticles as described in item 33 of the frequency range: the 4 U generator scans the first-voltage change of the change between the p« and the time-linear relationship, and the measurement is performed on the measurement-- The ion (4) is out of the particle, and its mass/charge ratio is linear with time. 36. The device for detecting nano particles according to claim 32, 〇578-A201〇2TWF2(N1); 〇7A -920505; david 31 1257705 further includes a second signal generator generating a oscillating number in the second ion w. Generating a sinusoidal function signal applied to the second ion trap for changing the electromagnetic field over time. The device for detecting nanoparticles as described in Item 2 is further provided. The ion supply source for the particles. 38. The apparatus for detecting nanoparticles according to claim 37, wherein the ion supply source comprises a matrix comprising particles. 39. The device for detecting nanoparticle according to claim 22, wherein the first-to-fourth (four) system is joined to the second ion, so that the 5th particle from the first-ion card is the second Ion trap capture. The method for detecting nano particles includes the following steps: applying a first-alternating voltage signal to a _th ion card containing charged particles from the frequency of the alternating voltage signal to periodically throw the charged The particle is suspected of: adding a second alternating voltage signal to a second ion trap that receives the T-electrode thrown from the first-ion trap; and a second ion trap based on a frequency of the first alternating voltage signal in. Scanning the second, the predetermined relationship of the frequency of the alternating voltage signal, such that the particles received by the second ion trap are limited to the method of detecting nanoparticles as described in claim 40, wherein the second alternating voltage is scanned The signal frequency maintains the solution parameter (qz) of the second ion solution unchanged. This is related to the particle received by the second ion taste. 42. The method of detecting nanoparticle according to claim 41, wherein the card parameter (qz) is proportional to the amplitude of the second alternating current signal, and the frequency of the second alternating voltage signal is Inverse ratio. A method for detecting nanoparticles as described in claim 4, further comprising periodically reducing the amplitude of the second alternating voltage signal, as described in claim 4, the method of detecting the nanoparticles of claim 4, To clear out the particles in the second ion trap. 44. A method of detecting nanoparticle, comprising the steps of: applying a control signal to a first ion trap such that the first ion trap periodically ejects the ion according to an ion mass/charge ratio; The second ion trap collects the ion thrown from the first ion; detects the fluorescent light excited by the ion in the second ion trap to generate a detection signal; and connects the detection signal and the control signal to determine a mass spectrum of the ions in the first ion trap. 45. The method of detecting nanoparticle as described in claim 44, further comprising directly irradiating a -substrate-laser light, inducing the occurrence of desorption and ionization reaction to generate the ion in the first ion trap . 46. The method of detecting nanoparticle according to claim 44, further comprising directly irradiating the laser light to the second ion to induce 47. As described in claim 44 The method for detecting nano particles further comprises adjusting the second material parameter to conform to the first-to-fourth (four): the second ion is fat for at least one specific time period ===== 】Package: adjusting the second ion The parameter of the card is the symbol = ^, and the first one has a -= 49. The method for detecting nano particles as described in claim 44, 0578-A20102TWF2 (N1); 07A-920505; david 33 1257705 This includes periodically clearing the ions from the second ion trap. 5. The method of detecting nanoparticle according to claim 49, wherein the signal of the amount of light emitted by the ion from the second ion is cleared from the second ion trap. 51. A device for measuring nanoparticle, comprising: a second ion trap, receiving ions periodically ejected from a first ion trap according to a mass/charge ratio; and a laser generator to generate a direct illumination a laser beam of the charged particle in the second ion trap; a light detector that detects a source of light emitted by the particle from the second ion; and a circuit that generates a control voltage applied to the second ion trap, such that The cation card clears the particle at a selected time, and the particle is ejected at least for a specific period of time such that the photodetector detects the glory emitted by the particle. 52. The apparatus for detecting nanoparticle according to claim 51, wherein the second ion card comprises a quadrupole ion having two cap electrodes (four) said 53. The device for detecting nano particles further includes a second generation voltage applied to the first ion trap to periodically eject the charged particles, the second control voltage having A frequency that is scanned from a first frequency to a second frequency. 0578-A201 〇2TWF2(N1); 07A-920505;david 34