CN109813591A - A kind of microbial concentration and purifying plant and microbial concentration and method of purification based on it - Google Patents
A kind of microbial concentration and purifying plant and microbial concentration and method of purification based on it Download PDFInfo
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- CN109813591A CN109813591A CN201910225715.XA CN201910225715A CN109813591A CN 109813591 A CN109813591 A CN 109813591A CN 201910225715 A CN201910225715 A CN 201910225715A CN 109813591 A CN109813591 A CN 109813591A
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Abstract
The invention discloses a kind of microbial concentration and purifying plants, the ultrasonic wave generating unit including sound fluid device and on the sound fluid device；The sound fluid device is equipped with micro-fluidic chamber, injection port and outlet, and the outlet includes at least the first outlet and the second outlet setting up and down, and the injection port is connected to the micro-fluidic chamber with the outlet；The ultrasonic wave generating unit is between the injection port and the outlet and is set to the micro-fluidic chamber upside or downside to generate ultrasonic wave upwards, the ultrasonic wave is standing wave, the micro-biological samples that the acoustic irradiation power that the standing wave generates flows through the micro-fluidic chamber form the microorganism stream of layering, and the micro-biological samples of different layers are flowed out along the different outlets respectively.Make microorganism purify microorganism by the effect of acoustic irradiation power, microorganism adhering is avoided to lose using the ultrasonic standing wave that ultrasonic wave generating unit generates, improve the rate of recovery, while by sample concentration, reducing cost.
Description
[technical field]
The present invention relates to microbial concentrations and purification technique field, and in particular to a kind of microbial concentration and purifying plant and
Based on its microbial concentration and method of purification.
[background technique]
In real life, drinking water is lower relative to the content of microorganisms in medical microbial test sample, for example posts
The standard detection of infested (flagellate, Cryptosporidium) is limited to 1/10 liters water；Escherichia coli standard detection is limited to 1CFU/100 milli
Rise water.In order to enable drinking water microorganism detection reaches standard detection limit, concentration and purification of the microorganism in Drinking Water Samples
It is the committed step carried out in drinking water microorganism detection process.
After existing drinking water microbial concentration collects concentration using the method that filter membrane filtering recoils with high pressure
Microbiological specimens, but microbiological recovery is low, since structure is complicated for filter membrane, most of microorganism is after filtration all in film
Upper adhesion.It is difficult effectively to backwash.The rate of recovery usually only 30%-40%.
It is existing drink water microorganism purification be immunoreacted using by biomolecular, corresponding microorganism is grabbed
It takes and passes through magnetic field and separated, because immunization needs to lead to higher cost using expensive antibody, and microorganism and antibody
In conjunction with needing the regular hour to be mixed, filter screen needs cleaning or replacement before concentration next time again, it is difficult to realize online
Continuous sample processing.
[summary of the invention]
The purpose of the present invention is to provide a kind of microbial concentration and purifying plants, solve existing microbial concentration technology and return
The problem of yield is low, existing microorganism purification technique is at high cost and is difficult to continuous online processing.
To solve the above problems, the present invention provides a kind of microbial concentration and purifying plant, including sound fluid device and set
Ultrasonic wave generating unit on the sound fluid device；The sound fluid device is equipped with micro-fluidic chamber, injection port and outlet,
The outlet includes at least the first outlet and the second outlet setting up and down, and the injection port is connected to the outlet
The micro-fluidic chamber；The ultrasonic wave generating unit is between the injection port and the outlet and is set to described micro-fluidic
On the upside of chamber or downside to generate ultrasonic wave upwards, and the ultrasonic wave is standing wave, and the acoustic irradiation power that the standing wave generates flows through
The micro-biological samples of the micro-fluidic chamber form the microorganism stream of layering, and the micro-biological samples of different layers are respectively described in different
Outlet outflow.
The ultrasonic wave generating unit is piezoelectric ceramics.
The upper side and lower side of the micro-fluidic chamber is two panels sheet glass disposed in parallel.
The area of the sheet glass is not less than 10cm2。
The distance between sheet glass described in two panels is 40-250 microns.
The wavelength of the standing wave is twice of distance between sheet glass described in two panels.
Compared with prior art, microbial concentration of the invention has the advantage that with purifying plant
1, the ultrasonic standing wave that microbial concentration of the invention and purifying plant are generated using ultrasonic wave generating unit is to micro- life
Object carries out contactless manipulation, the sound wave that the acoustic irradiation power that ultrasonic wave generates makes itself that there is the microorganism of different characteristics to be subject to
Radiant force is different, and is gathered in the space of micro-fluidic intracavitary different height under this force, to pass through different outlets
Outflow sound fluid device, obtains micro-biological samples after purification, and in this process, it avoids micro- caused by using filter screen
Bioadhesion loss, improves the rate of recovery of microbial concentration technology；
2, while being purified, the water sample for removing microorganism is also divided for microbial concentration of the invention and purifying plant
It separates out, micro-biological samples is concentrated, do not need to be immunoreacted using antibody and microorganism, reduce and be purified to
This；
As long as 3, the sound fluid device of microbial concentration of the invention and purifying plant does not damage, so that it may it works always,
When needing to be concentrated new micro-biological samples, the micro-fluidic chamber of on-line cleaning sound fluid device is only needed and into and out of after sample mouth
It is concentrated and is purified, solve the problems, such as that the prior art is difficult to continuous online processing sample.
The present invention also provides a kind of microbial concentration and method of purification based on mentioned microorganism concentration and purifying plant, tools
Body step includes:
(a) microorganism stream sample is introduced into the injection port；
(b) start the ultrasonic wave generating unit and generate ultrasonic wave, the ultrasonic wave is standing wave；
(c) when the microorganism stream sample flows through the micro-fluidic chamber, the acoustic irradiation power of the standing wave is to difference
Microorganism form different thrust so that different microorganisms forms layering in the micro-fluidic intracavitary different height space
Microorganism stream；
(d) the microorganism stream of different layers is flowed out from the outlet of different layers respectively.
The calculation formula of the acoustic irradiation power is as follows:
Frad=1/2 (π Φ ka3Eac),
In above formula, π is pi, and Φ is acoustics reduced factor, and k is wave vector, and a is microorganism diameter, EacFor sound wave energy
Force density.
Flow velocity of the microorganism stream sample in the sound fluid device is not less than 100ml/min.
The sound fluid device be equipped with it is multiple, be connected with each other between multiple sound fluid devices, and two adjacent institutes
It states in sound fluid device, the injection port of the posterior sound fluid device connects the sample out of the preceding sound fluid device
Mouthful.
Of the invention is had the advantage that based on mentioned microorganism concentration and the microbial concentration of purifying plant with method of purification
Microbial concentration of the invention and method of purification improve the concentration and recovery rate of microorganism, it can be achieved that continuous concentration with
No replacement is required while purification equipment and consumptive material, reduction processing cost.
[Detailed description of the invention]
Fig. 1 is the schematic perspective view of sound fluid device described in the embodiment of the present invention.
Fig. 2 is the operation principle schematic diagram of sound fluid device described in the embodiment of the present invention.
Fig. 3 is microorganism stress schematic diagram described in the embodiment of the present invention.
Fig. 4 is micrograph before the concentration of microorganism described in the embodiment of the present invention.
Fig. 5 is micrograph after microbial concentration described in the embodiment of the present invention.
[specific embodiment]
Following will be combined with the drawings in the embodiments of the present invention, carries out clear, complete description to technical solution of the present invention,
Obviously, the described embodiments are merely a part of the embodiments of the present invention, instead of all the embodiments.Based in the present invention
Embodiment, every other embodiment obtained by those of ordinary skill in the art without making creative efforts, all
It belongs to the scope of protection of the present invention.
Embodiment: attached drawing 1 is please referred to attached drawing 5, the present embodiment provides a kind of microbial concentration and purifying plants, including sound
Fluid device 1 and the ultrasonic wave generating unit 15 on the sound fluid device 1, sound fluid device 1 are mainly used for micro- life
Object is concentrated and is purified；The sound fluid device 1 is equipped with micro-fluidic chamber 11, injection port 12 and outlet 13, the injection port
12 are connected to the micro-fluidic chamber 11 with the outlet 13, form the negotiable environment of microorganism stream sample；It is described micro-
Main place of the flow control chamber 11 as concentration and purification, so that the microorganism with different characteristics is produced by ultrasonic vocalization unit
Raw acoustic irradiation power is different, and is gathered in micro-fluidic intracavitary different spaces height under this force, convenient for separating and mentioning
Pure, the different characteristics that microorganism has refer to the inherent parameters such as its granular size, density, compressible；Based on this, pass through control
Flow velocity, micro-fluidic cavity configuration size and frequency of sound wave, the parameters such as intensity, can control different microorganisms after micro-fluidic chamber with
Different flow velocitys is by different outlets, to reach the concentration and purification of microorganism；
The injection port 12 is needed for inputting needs and being concentrated with isolated microorganism stream sample in the present embodiment
Being concentrated with isolated microorganism stream sample is the drinking water containing microorganism, includes the first microorganism 18 and the second microorganism
19；The outlet 13 includes at least the first outlet and the second outlet setting up and down, certainly, is concentrated for actual needs
The type and quantity of microorganism, can be set multiple outlets.The outlet 13 is for flowing out the water for removing certain microorganism
Sample and containing certain by the water sample of separate microorganism, such as contain the water sample of the first microorganism 18 and the second microorganism 19, in this implementation
In example, there are four the outlets, for the ease of indicating that the substance that different outlets are flowed out is different, in addition to one of them goes out sample
It is outer that mouth uses label 13 to mark, and is left three outlets using the different marks of label 14,16,17, outlet 16 is to set upper
The first outlet set, outlet 14 are the second outlet in lower setting, and the outflow of outlet 13 is to remove certain microorganism
Water sample, what outlet 17 flowed out is also to remove the water sample of certain microorganism, and the outflow of outlet 14 is that the second microorganism 19 contains
High water sample is measured, the outflow of outlet 16 is the high water sample of 18 content of the first microorganism；Particularly, in the present embodiment, described
Injection port 12 and the outlet 17 are located at one end of the micro-fluidic chamber 11, and the outlet 13,14,16 is located at the miniflow
The other end of chamber is controlled, setting in this way is convenient for being separated off the water sample of certain microorganism from outlet 17, sees attached drawing 1.
The ultrasonic wave generating unit 15 is between the injection port 12 and the outlet 13 and is set to the miniflow
11 upside of chamber or downside are controlled to generate ultrasonic wave upwards, specifically, the direction of the ultrasonic wave acts on the micro-fluidic chamber
It is axial, microorganism can be made in the intracavitary microorganism stream for being formed layering by acoustic irradiation power of the flow control；In the present embodiment
In, the downside of the micro-fluidic chamber is arranged in the ultrasonic wave generating unit, see attached drawing 1, certainly, the ultrasonic wave generating unit
Also it can be set in the upside of the micro-fluidic chamber, corresponding change need to be made to the position of the outlet only convenient for micro- life of layering
Logistics goes out, and the ultrasonic wave generating unit 15 is piezoelectric ceramics, i.e., the described ultrasonic vocalization unit is produced as ultrasound source
Raw ultrasonic wave, the ultrasonic wave are standing wave, that is, the ultrasonic wave generated is in the micro-fluidic intracavitary formation standing wave, the side of the standing wave
To the axial direction perpendicular to the microorganism sound fluid device 1, so that the microorganism can gather under the action of acoustic irradiation power
For collection in the space of same level namely the micro-fluidic intracavitary sustained height, the wavelength X of the standing wave is described in two panels
Twice of 111 distance of sheet glass, the i.e. half of the distance between sheet glass described in two panels be equal to λ/4, so as to microorganism by
It can move, be scattered at the microorganism in microorganism water sample to micro-fluidic intracavitary structure centre when the effect of acoustic irradiation power
In micro-fluidic intracavitary, the microorganism close to two blocks of sheet glass is respectively by incident ultrasound wave and the acoustic irradiation power reflected ultrasonic wave
FradEffect；See attached drawing 3, because sound wave be it is sightless, be all represented by dashed line herein, n in attached drawing 31、n2Respectively indicate difference
The incident ultrasound wave waveform of phase, n1、n2Reflect ultrasonic wave and n1、n2Symmetrical about line L, monolithic piezoelectric ceramics all generate super
Sound wave, dotted line is pointed out in figure only four therein, only illustrates standing wave.
The size of acoustic irradiation power can be calculated by following formula:
Frad=1/2 (π Φ ka3Eac),
In above formula, π is pi, and Φ is acoustics reduced factor, and k is wave vector, and a is microorganism diameter, EacFor sound wave energy
Force density.
The micro-biological samples that the acoustic irradiation power that the standing wave generates flows through the micro-fluidic chamber 11 form the micro- of layering
Biology stream, when microorganism enters cavity, in acoustic irradiation power FradUnder the action of respectively towards micro-fluidic chamber structure centre move
Dynamic, at pressure node m, what microorganism here was subject to enter, reflected acoustic wave radiant force FradIt is equal in magnitude contrary,
It cancels out each other, thus the thrust that microorganism is only travelled forward by water flow, such germs collect are occupied at one layer of microorganism stream
The microorganism stream in the certain altitude space of micro-fluidic chamber, different layers is flowed out along the different outlets 13 respectively, i.e., from acoustic streaming
Flowing out on body device with microorganism outlet similar in microorganism stream height, sees attached drawing 2, separates the quasi-microorganism to reach
Purpose.
The upper side and lower side of the micro-fluidic chamber 11 is two panels sheet glass 111 disposed in parallel, and glass sheet surface is smooth, micro-
Biology is not easy to be adhered to glass surface.
The area of the sheet glass 111 is not less than 10cm2, the wavelength of the standing wave is between sheet glass 111 described in two panels
Twice of distance；The distance between sheet glass 111 described in two panels is 40-250 microns, and the area of the sheet glass 111 is greater than
Or it is equal to 10cm2, the area of sheet glass described herein is more than or equal to 10cm2With the fixation between sheet glass 111 described in two panels
Distance so that micro-fluidic chamber two sides glass formed a plate shaped sound fluidic structures, can allow processing drinking water flow velocity
It improves, further increases concentration and recovery efficiency.Certainly, the flow velocity of water can be further promoted by area increased, while corresponding
Increase the size of piezoelectric ceramics, and then improves efficiency.
The ultrasonic standing wave that the microbial concentration and purifying plant of the present embodiment are generated using ultrasonic wave generating unit is to micro- life
Object carries out contactless manipulation, and the acoustic irradiation power that ultrasonic wave generates makes itself have the microorganism stress of different characteristics different,
And it is gathered in the space of micro-fluidic intracavitary different height under this force, to leave acoustic streaming body device by different outlets
Part obtains micro-biological samples after purification, and in this process, avoid the damage of microorganism adhering caused by using filter screen
It loses, improves the rate of recovery of microbial concentration technology；Microbial concentration and purifying plant of the invention while being purified,
The water sample for removing microorganism is also separated, and micro-biological samples are concentrated, do not need using antibody and microorganism into
Row immune response, reduces cost for purification；As long as the sound fluid device of microbial concentration and purifying plant of the invention does not damage,
Can work always, when needing to be concentrated new micro-biological samples, only need on-line cleaning sound fluid device micro-fluidic chamber and
It can be concentrated and purify after into and out of sample mouth, solve the problems, such as that the prior art is difficult to continuous online processing sample.
The ultrasonic standing wave that microbial concentration and purifying plant of the invention is generated using ultrasonic wave generating unit is to microorganism
Contactless manipulation is carried out, the acoustic irradiation power that ultrasonic wave generates makes itself have the microorganism stress of different characteristics different, and
It is gathered in the space of micro-fluidic intracavitary different height under this force, to leave acoustic streaming body device by different outlets
Part obtains micro-biological samples after purification, and in this process, avoid the damage of microorganism adhering caused by using filter screen
It loses, improves the rate of recovery of microbial concentration technology；Microbial concentration and purifying plant of the invention while being purified,
The water sample for removing microorganism is also separated, and micro-biological samples are concentrated, do not need using antibody and microorganism into
Row immune response, reduces cost for purification；As long as the sound fluid device of microbial concentration and purifying plant of the invention does not damage,
Can work always, when needing to be concentrated new micro-biological samples, only need on-line cleaning sound fluid device micro-fluidic chamber and
It can be concentrated and purify after into and out of sample mouth, solve the problems, such as that the prior art is difficult to continuous online processing sample.
The present embodiment also provides a kind of microbial concentration and method of purification based on mentioned microorganism concentration and purifying plant,
Specific steps include:
(a) microorganism stream sample is introduced into the injection port 12, to carry out the work of next step separating-purifying；
(b) start the ultrasonic wave generating unit 15 and generate ultrasonic wave, the ultrasonic wave is standing wave, and the standing wave is to micro- life
Object generates acoustic irradiation power；
(c) when the microorganism stream sample flows through the micro-fluidic chamber 11, the micro-fluidic chamber is dense as microorganism
The main place of contracting and purification, the acoustic irradiation power of the standing wave forms different thrust to different microorganisms, due to difference
The microorganism of type has different characteristics, and the different characteristics that microorganism has refer to its granular size, density, compressible etc.
Inherent parameters；The effect of acoustic irradiation power by standing wave is also just different, so that different microorganisms is in the micro-fluidic chamber 11
Interior different height space forms the microorganism stream of layering, such as above-mentioned first microorganism 18 and the second microorganism 19, and first is micro-
The acoustic irradiation power that biology 18 is subject to is larger, and the structure centre towards micro-fluidic chamber moves, and is continually entering the micro- of micro-fluidic chamber
Under the promotion of biofluid, forms the high microorganism of 18 content of the first microorganism and flow, and the sound wave spoke that the second microorganism 19 is subject to
It is smaller to penetrate power, still close to the sheet glass flow forward, forms the high microorganism stream of 19 content of the second microorganism；
(d) the microorganism stream of different layers is flowed out from the outlet 13 of different layers respectively, and microorganisms different in this way is just
The purpose for realizing separating-purifying, while being purified, remove certain microorganism drink water sample from outlet 13 and go out
Sample mouth 17 flows out, and micro-biological samples are concentrated.
The calculation formula of the acoustic irradiation power is as follows:
Frad=1/2 (π Φ ka3Eac),
In above formula, π is pi, and Φ is acoustics reduced factor, and k is wave vector, and a is microorganism diameter, EacFor sound wave energy
Force density.
Further, in order to improve the efficiency of microbial concentration and method of purification, the microorganism stream sample is described
Flow velocity in sound fluid device 1 is not less than 100ml/min, and the sample flow in sound fluid device 1 is bigger, the concentration of microorganism
Faster with the speed of purification, efficiency is higher.
Further, in order to improve microbial concentration and method of purification purification microorganism purity, can also be by certain
The microorganism of kind initial gross separation carries out further being concentrated and separating again, and the specific method is as follows, and the sound fluid device 1 is equipped with more
It is a, it is connected with each other between multiple sound fluid devices 1, and in two adjacent sound fluid devices 1, the posterior sound
The injection port 12 of fluid device 1 connects the outlet 13 of the preceding sound fluid device 1, utilizes micro- life of initial gross separation
The different characteristics of object are further separated using the sound fluid device of corresponding construction, keep the purity of microorganism further
It improves.
The microbial concentration and refining effect of the present embodiment, are shown in attached drawing 4 and attached drawing 5.Attached drawing 4 is described in the embodiment of the present invention
Microorganism concentration before micrograph, in the present embodiment the microorganism be Escherichia coli, i.e., the described attached drawing 4 is Escherichia coli
Micrograph before sample concentration, A is the Escherichia coli before concentration in figure；Attached drawing 5 is microorganism described in the embodiment of the present invention
Micrograph after concentration, the microorganism is Escherichia coli in the present embodiment, and the attached drawing 5 is after E. coli SampLes are concentrated
Micrograph, B is the Escherichia coli after concentration in figure；Due to detection Escherichia coli quantity when, Escherichia coli and fluorescence immunoassay are anti-
Precursor reactant issues blue-fluorescence under the microscope, may determine that the concentration effect of Escherichia coli from blue-fluorescence points and closeness
Fruit；When blue-fluorescence is brighter, phosphor dot is more intensive, illustrate that Escherichia coli quantity is more, concentrated effect is better, from 4 He of attached drawing
The effect of 5 Escherichia coli of attached drawing concentration front and back sees that the blue-fluorescence points of B are significantly more than the phosphor dot of A in attached drawing 4 in attached drawing 5
It counts, the blue-fluorescence of blue-fluorescence also obviously made a farfetched comparison in Fig. 4 is bright in attached drawing 5, and the Escherichia coli after illustrating concentration are more dense, dense
Contracting effect is more preferable.
The microbial concentration and method of purification of the present embodiment improve the concentration and recovery rate of microorganism, it can be achieved that continuous concentration
No replacement is required while with purification equipment and consumptive material, reduction processing cost；Multiple sound fluid devices interconnected further mention
The high purity of microorganism.
Microbial concentration of the invention and method of purification improve the concentration and recovery rate of microorganism, it can be achieved that continuous concentration with
No replacement is required while purification equipment and consumptive material, reduction processing cost；Multiple sound fluid devices interconnected further increase
The purity of microorganism.
For the ordinary skill in the art, it is possible to understand that the case where not departing from the principle and spirit of the invention
Under these embodiments can be carried out with a variety of change, modification, replacement and modification, the scope of the present invention by appended claims and its
Equivalent limits.
Claims (10)
1. a kind of microbial concentration and purifying plant, it is characterised in that: including sound fluid device (1) and be set to the acoustic streaming body device
Ultrasonic wave generating unit (15) on part (1)；The sound fluid device (1) is equipped with micro-fluidic chamber (11), injection port (12) and goes out
Sample mouth (13), the outlet (13) include at least the first outlet and the second outlet setting up and down, the injection port
(12) the micro-fluidic chamber (11) is connected to the outlet (13)；The ultrasonic wave generating unit (15) is located at the injection port
(12) described to generate ultrasonic wave upwards between the outlet (13) and set on micro-fluidic chamber (11) upside or downside
Ultrasonic wave is standing wave, and the micro-biological samples that the acoustic irradiation power that the standing wave generates flows through the micro-fluidic chamber (11), which are formed, divides
The microorganism stream of the microorganism stream of layer, different layers is flowed out along the different outlets (13) respectively.
2. microbial concentration according to claim 1 and purifying plant, it is characterised in that: the ultrasonic device (15) is
Piezoelectric ceramics.
3. microbial concentration according to claim 2 and purifying plant, it is characterised in that: the micro-fluidic chamber (11) it is upper
Side and downside are two panels sheet glass disposed in parallel (111).
4. microbial concentration according to claim 3 and purifying plant, it is characterised in that: the face of the sheet glass (111)
Product is not less than 10cm2。
5. microbial concentration according to claim 4 and purifying plant, it is characterised in that: sheet glass described in two panels
It the distance between (111) is 40-250 microns.
6. microbial concentration according to claim 5 and purifying plant, it is characterised in that: the wavelength of the standing wave is two panels
Twice of distance between the sheet glass (111).
7. a kind of microbial concentration and method of purification based on claim 1~6 any one described device, which is characterized in that
Specific steps include:
(a) microorganism stream sample is introduced into the injection port (12)；
(b) start the ultrasonic device (15) and generate ultrasonic wave, the ultrasonic wave is standing wave；
(c) when the microorganism stream sample flows through micro-fluidic chamber (11), the acoustic irradiation power of the standing wave is to difference
Microorganism form different thrust so that different microorganisms height space different in the micro-fluidic chamber (11) is formed
The microorganism of layering is flowed；
(d) the microorganism stream of different layers is flowed out from the outlet (13) of different layers respectively.
8. microbial concentration according to claim 7 and method of purification, it is characterised in that: the calculating of the acoustic irradiation power
Formula is as follows:
Frad=1/2 (π Φ ka3Eac),
In above formula, π is pi, and Φ is acoustics reduced factor, and k is wave vector, and a is microorganism diameter, EacIt is close for sound wave ability
Degree.
9. microbial concentration according to claim 7 and method of purification, it is characterised in that: the microorganism stream sample exists
Flow velocity in the sound fluid device (1) is not less than 100ml/min.
10. microbial concentration according to claim 7 and method of purification, it is characterised in that: the sound fluid device (1) sets
Have multiple, is connected with each other between multiple sound fluid devices (1), and in two adjacent sound fluid devices (1), rear
The injection port (12) of the sound fluid device (1) connect the outlet (13) of the preceding sound fluid device (1).
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