CN107841526A - A kind of malic dehydrogenase diagnostic kit - Google Patents
A kind of malic dehydrogenase diagnostic kit Download PDFInfo
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- CN107841526A CN107841526A CN201711137382.2A CN201711137382A CN107841526A CN 107841526 A CN107841526 A CN 107841526A CN 201711137382 A CN201711137382 A CN 201711137382A CN 107841526 A CN107841526 A CN 107841526A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
- C12Q1/32—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase involving dehydrogenase
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
Abstract
The invention belongs to technical field of biological, discloses a kind of malic dehydrogenase diagnostic kit, is provided with the first casing and the second casing mutually pasted；The middle part riveting of first casing has rotating shaft, and rotating disk is set with the rotating shaft, some detection boxes are provided with the outside of the rotating disk；Medicine box is provided with the inside of the detection box；Multiple injection devices are installed with second casing, detection cell is welded with the top of first casing, the upper end of the detection cell is set with medicine addition box.The kit sensitiveness is high, and high specificity is reproducible.It is easy to operate due to its stable reagent, easy to maintain, the factor such as more objective is as a result judged, suitable for determining the malate dehydrogenase activity in different biological samples, such as cell and tissue culture supernatant and the mitochondria of purifying gained.
Description
Technical field
The invention belongs to technical field of biological, more particularly to a kind of malic dehydrogenase diagnostic kit.
Background technology
Malic dehydrogenase (MalateDehydrogenase, MDH) is one of key enzyme of synthesizing apple acid, is catalyzed apple
The mutual conversion of tartaric acid and oxaloacetic acid (OAA), participate in the oxidation of numerous physiological metabolism approach such as TCA circulations C4 recycled fatty acids
Respiration nitrogen assimilation etc., therefore, MDH play an important role in the growing of plant, and are widely present in mitochondria, thin
It is a kind of enzyme in tricarboxylic acid cycle on bacterium cell membrane, because the source of enzyme is different, its some property is also not quite similar.MDH exists
Important role is play in a variety of physiological activities of cell, includes energetic supersession, the malate aspartate shuttle system of mitochondria
System, active oxygen metabolism and disease resistance etc..According to different coenzyme specificities, MDH points rely on for the NAD- MDH relied on and NADP-
MDH, generally comprise only NAD-MDH in bacterium, in eukaryotic, NAD-MDH is distributed in cytoplasm and mitochondria.
Malic dehydrogenase (MDH) is present in all organisms, is one of key enzyme of biological glycometabolism, can be catalyzed
Reversible transformation between malic acid and oxaloacetic acid.Acute myocardial infarction AMI, malignant tumour, traumatic shock, kidney trouble, class wind
Various diseases of wet arthritis, hematological system etc., it is seen that MDH increased activities.Viral hepatitis serum MDH gross activities and apple
Acidohydrogenase isodynamic enzyme M (M-MDH) elevation amplitude is related to the seriousness of the state of an illness, and early stage measure M-MDH meanings are bigger,
When oxyhepatitis necroses, Chances of Malic Dehydrogenase Isoenzyme C (C-MDH) rises are more prominent, and have prognosis meaning to the state of an illness
Justice.But the diagnosis for malic dehydrogenase is excessively cumbersome, Check-Out Time length, operate cumbersome.
In summary, the problem of prior art is present be：Diagnosis of the existing kit for malic dehydrogenase is excessively
It is cumbersome, Check-Out Time length, operate cumbersome.
The content of the invention
The problem of existing for prior art, the invention provides a kind of malic dehydrogenase diagnostic kit.
The present invention be achieved in that the malic dehydrogenase diagnostic kit be provided with the first casing for mutually pasting and
Second casing；The middle part riveting of first casing has rotating shaft, and rotating disk is set with the rotating shaft, and the outside of the rotating disk is set
There are some detection boxes；Medicine box is provided with the inside of the detection box；
Multiple injection devices are installed with second casing, detection cell is welded with the top of first casing, it is described
The upper end of detection cell is set with medicine addition box.
Further, the inside of the detection box is filled with hemostasis disinfection agent of chlorine dioxide solution, disappears in hemostasis chlorine dioxide
In toxic agent solution, comprising 0.1~50u/ml with local quick-acting haemostatic powder effect hemostasis biology enzyme, 0.01~1% with
The disinfection agent of chlorine dioxide of the concentration of disinfection effect and 1~100mg/ml protein protective agent (nitrine can be reached
Sodium).
Further, the rear end of the rotating disk is provided with drive device, and supply unit is connected with the drive device, described
Supply unit includes：Rechargeable battery, current diverter, multiple current comparators and multiple delivery outlets；
The input of current diverter is connected with rechargeable battery, and the output end of current diverter is defeated with each current comparator
Enter end connection, the output end of each current comparator is respectively connecting to corresponding delivery outlet；
Also include multiple electronic switches, each electronic switch be separately positioned on corresponding between current comparator and delivery outlet,
Both ends are connected with current comparator and delivery outlet respectively；The quantity of current comparator, delivery outlet and electronic switch is four, point
It is not：First current comparator, the second current comparator, the 3rd current comparator, the 4th current comparator, the first delivery outlet,
Second delivery outlet, the 3rd delivery outlet, the 4th delivery outlet, the first electronic switch, the second electronic switch, the 3rd electronic switch and the 4th
Electronic switch, the output end of current diverter respectively with the first current comparator, the second current comparator, the 3rd current comparator
Connected with the input of the 4th current comparator, the output end of the first current comparator is exported by the first electronic switch and first
Mouth connection, the output end of the second current comparator are connected by the second electronic switch with the second delivery outlet, the 3rd current comparator
Output end be connected by the 3rd electronic switch with the 3rd delivery outlet, the output end of the 4th current comparator passes through the 4th electronic cutting
Pass is connected with the 4th delivery outlet；
The rated current of power supply unit is I, and the output current of the first delivery outlet is I1, the output current of the second delivery outlet
For I2, the output current of the 3rd delivery outlet is I3, the output current of the 4th delivery outlet is I4；Correspondingly, first current comparator
A reference value is I, and a reference value of the second comparator is I-I1, a reference value of the 3rd comparator is I-I1-I2, the base of the 4th comparator
Quasi- value is I-I1-I2-I3。
Further, the injection device uses electric injection device, the injecting method of the electric injection device include with
Lower step：
Step 1: the culture dish containing cell is positioned on locating platform first, while sticking pin and injection needle are divided
Do not tilt down and be fixed on left adjustable plate and right adjustable plate, simultaneous camera detects the image feelings of inverted microscope in real time
Condition, and signal is passed into computer；
Step 2: according to the feedback information of video camera, computer export signal feeding bit platform controller, locating platform control
Device control locating platform processed is all around moved in the horizontal direction so that culture dish is correspondingly arranged with inverted microscope；
It is as follows that the video camera carries out rectangle partitioning algorithm specific method in a scanning area to region of variation：
(1) image transmitting terminal obtains the resolution ratio of screen first, obtains 0~C of scope of column scan and the scope 0 of row scanning
~R；
(2) data of current frame image conservation zone are saved in previous frame image buffering area by transmitting terminal；Intercept and capture current screen
Curtain bitmap data is simultaneously stored in current frame image buffering area；
(3) it is (0,0) that transmitting terminal initializes change rectangular area top left co-ordinate and bottom right angular coordinate first, scanning next time
Starting point coordinate is (0,0), and row is unchanged to be identified as true, updates the scope of column scan and the scope of row scanning；
(4) judge whether to be expert in scanning range, do not exist, jump to (10)；
(5) judge whether in the range of column scan, do not exist, jump to (8)；Use in the range of column scan and directly compare every row
Current sampling point is detected compared with method；Value is different, and the unchanged mark of row is arranged into false first, then judge whether be
The first change sampled point detected, is the top left co-ordinate using sample point coordinate as change rectangular area, is not first
Individual change sampled point, relatively and maximum is taken to be sat as the new rectangle lower right corner on the coordinate of the coordinate in the rectangle lower right corner and the point
Mark, then judge whether the sampled point is first change sampled point of one's own profession, it is just by the ordinate of the sampled point with rectangle upper left
The ordinate at angle is compared and takes the top left co-ordinate of minimum value more new change rectangular area；It is worth identical, it is necessary to judge to go nothing
Change is identified whether as false, if false, starting point that record coordinate scans as next time detects it is that last row is adopted
Sampling point, the starting point using last row sample point coordinate as scanning next time, is jumped to (7)；
(6) row coordinate is moved to right N row, jumps to step 5 and detect next sampled point；
(7) one's own profession detection is finished, and the next sweep starting point of the next sweep starting point coordinate of one's own profession and lastrow record is sat
Mark compares, and takes maximum to add 1 as new next sweep starting point coordinate, line number, jump to step 4 and from the beginning opened from next line
Beginning is from left to right detected；
(8) judge that row is unchanged and identify whether that for true and change rectangular area top left co-ordinate be not (0,0), be not
True, line number add 1, jump to (4)；It is true, then shows that full line without different pixels, has obtained the rectangular area of a change
Block；Obtained change rectangular area block upper left corner ordinate be moved to the left N row, lower right corner ordinate move right N arrange with comprising
Image boundary information；
(9) the change rectangular area coordinate and corresponding next sweep starting point coordinate, judgement that record detects work as forefront
The scope of scanning whether 0~C and row scanning scope whether 0~R, be, set mark show the change rectangle that current detection goes out
Area identification is to detect for the first time, and then line number adds 1 to jump to the rectangle that (4) detect next change since next line
Region unit；Until detecting the scope beyond row scanning；
(10) after this detection, next sweep starting point all in this detection is handled, calculates next time
The set of scanning range；Whether the ordinate for first checking for this first next sweep starting point detected is adopted than last row
The ordinate of sampling point is small, is not, the region detection is completed, and detects the ordinate of next next sweep starting point；It is, with for the first time
The abscissa in the change rectangular area upper left corner detected is abscissa, is scanned related next time with currently changing rectangular area
The ordinate of point coordinates is ordinate, generates the top left co-ordinate of a next scanning range；With the change detected for the first time
The abscissa in the rectangular area lower right corner is abscissa, and a next scanning range is generated by ordinate of the maximum number of column C of screen
Bottom right angular coordinate；Then handle second next sweep starting point, until next sweep starting point all in this detection all by
Untill processing；
(11) scanning area all in next scanning range set is detected, is primarily based in next scanning range set the
The scope of the width and height of one scanning area, raw row scan and column scan, repeat (3) to (10) and detect first scanning
The rectangular area block changed in region, second scanning area is then handled, is swept until all in next scanning range set
Retouch region all be detected untill；
(12) (10) are repeated and arrives (11), the change rectangular area block of scanning range next time is obtained, until all next times
The ordinate of sweep starting point is more than or equal to the ordinate of last row sampled point, and whole screen detection finishes；
(13) the not overlapping rectangles region for the area minimum that all two field pictures change relative to previous frame image has been obtained
Set, check the rectangular area in the set, two rectangle its upper left corner ordinates are identical with lower right corner ordinate, and one
The lower right corner abscissa of rectangle is adjacent with another rectangle upper left corner abscissa, merges into a rectangle, and then recompression is concurrent
View data that the set of rectangular area is included and respective coordinates are sent to client；
(14) image-receptive end will be based on each rectangular region image data after the data decompression of reception and respective coordinates are whole
It is bonded in previous frame image and shows；
(15) (2) are repeated every the T seconds and arrives (14), according to the difference of application scenarios and the requirement of bandwidth, interval time T is done
Adjustment；
Step 3: auto-focusing is carried out to the cell in culture dish by inverted microscope, target identification finds cell；
Step 4: computer export signal gives adjustable plate controller, adjustable plate controller drives left adjustable plate operation so that
Sticking pin and cells contacting, while start linear electrical platform and drive syringe movement to produce negative pressure so that sticking pin is held carefully
Born of the same parents；
The signal r that the adjustable plate controller receiveskIt is expressed as：
In formula, A is signal amplitude, is unknown constant in a burst frame；foFor carrier wave frequency deviation, in a burst frame
For unknown constant；TsFor sampling period, foTsFor normalized carrier frequency shift；anFor QPSK modulation datas；θ0For skew,
It is unknown constant in one burst frame；G sends pulse with receiving the product of matched filter impulse function；nkFor multiple Gauss white noise
Sound, obey N (0, σ2) distribution；Timing Complete Synchronization during ε=0, otherwise timing is not synchronous；K is time sequence number, and N is over-sampling times
Number；rkThere is 10dB dynamic range；The demodulation method main task of the short preamble burst signal of low signal-to-noise ratio is from rkMiddle recovery
Go out to send data；
The computer is to signal time-frequency domain matrixPre-processed, specific bag
Include following two step：
The first step is rightCarry out low energy to pre-process, i.e., in each sampling instant
P, willValue of the amplitude less than thresholding ε is set to 0, and is obtained
Thresholding ε setting can determine according to the average energy of reception signal；
Second step, the time-frequency numeric field data of p moment (p=0,1,2 ... P-1) non-zero is found out, used
Represent, whereinRepresent the response of p moment time-frequencyCorresponding frequency indices, right when non-zero
The normalization pretreatment of these non-zeros, obtains pretreated vectorial b (p, q)=[b1(p,q),b2(p,q),…,bM(p,
q)]T, wherein
Step 5: the fixed injection needle of aft ramp controller regulation right adjustable plate so that injection needle connects cytoproximal
Cell membrane, now the piezoelectric ceramics power supply give the piezoelectric ceramic drive signal, piezoelectric ceramics produces vibration, realizes cell
Rupture of membranes, piezoelectric ceramics power supply is closed afterwards；
Step 6: computer export signal gives injection pump controller, injection pump controller control injection pump operation, pass through note
Penetrate pin and tissue is injected into cell desired position, complete the microinjection of cell.
Further, the adding method thereof of the medicine addition box comprises the following steps：
Step 1: receive dosing instruction；
Step 2: being instructed according to the dosing, the scarce medicine information that need to carry out feeding operations is searched by display device, if
Lookup result demonstrates the need for adding a certain or a variety of medicines, then performs next step；If lookup result shows to appoint without addition
Which kind of similar drug, then terminate dosing flow, and user need not carry out feeding operations；
Step 3: confirm the scarce medicine information of the first medicine that need to be added；
Step 4: according to the scarce medicine information of the first medicine, the first medicine is added in manipulator, and by addition
Medicine information is input in database control centre；
Step 5: the first medicine that manipulator will receive, is transported to set in advance with placing to be added first
The first dosing region corresponding to kind medicine；
Step 6: the Quantity of drugs that manipulator lacks according to the first dosing tank corresponding with the first dosing area, by the first
Medicine is added to the first dosing tank；
Step 7: the first dosing tank by the quantity of the first medicine received, feeds back to database control centre, data
Scarce medicine information in storehouse control centre more new system；
Step 8: judging whether manipulator also needs to move to other dosing areas adds the first medicine, if it is, machine
Tool hands movement adds the first medicine to other dosing areas, if it is not required, then manipulator returns to initial loading position.
Advantages of the present invention and good effect are：The kit sensitiveness is high, and high specificity is reproducible.Due to its examination
Agent is stable, easy to maintain, easy to operate, as a result judges the factor such as more objective, suitable for determining the malic acid in different biological samples
Dehydrogenase activity, such as cell and tissue culture supernatant and the mitochondria of purifying gained.
Brief description of the drawings
Fig. 1 is the structural representation of malic dehydrogenase diagnostic kit provided in an embodiment of the present invention；
Fig. 2 is the circuit theory diagrams of supply unit provided in an embodiment of the present invention；
In figure：1st, the first casing；2nd, the second casing；3rd, rotating shaft；4th, box is detected；5th, Medicine box；6th, injection device；7th, detect
Pond；8th, medicine addition box；9th, rotating disk.
Embodiment
In order to further understand the content, features and effects of the present invention, hereby enumerating following examples, and coordinate accompanying drawing
Describe in detail as follows.
1 pair of structure of the invention is explained in detail below in conjunction with the accompanying drawings.
The malic dehydrogenase diagnostic kit is provided with the first casing and the second casing mutually pasted；First case
The middle part riveting of body has rotating shaft, and rotating disk is set with the rotating shaft, some detection boxes are provided with the outside of the rotating disk；The inspection
Survey on the inside of box and be provided with Medicine box；
Multiple injection devices are installed with second casing, detection cell is welded with the top of first casing, it is described
The upper end of detection cell is set with medicine addition box.
Further, the inside of the detection box is filled with hemostasis disinfection agent of chlorine dioxide solution, disappears in hemostasis chlorine dioxide
In toxic agent solution, comprising 0.1~50u/ml with local quick-acting haemostatic powder effect hemostasis biology enzyme, 0.01~1% with
The disinfection agent of chlorine dioxide of the concentration of disinfection effect and 1~100mg/ml protein protective agent (nitrine can be reached
Sodium).
Further, the rear end of the rotating disk is provided with drive device, and supply unit is connected with the drive device, described
Supply unit includes：Rechargeable battery, current diverter, multiple current comparators and multiple delivery outlets；
The input of current diverter is connected with rechargeable battery, and the output end of current diverter is defeated with each current comparator
Enter end connection, the output end of each current comparator is respectively connecting to corresponding delivery outlet；
Also include multiple electronic switches, each electronic switch be separately positioned on corresponding between current comparator and delivery outlet,
Both ends are connected with current comparator and delivery outlet respectively；The quantity of current comparator, delivery outlet and electronic switch is four, point
It is not：First current comparator, the second current comparator, the 3rd current comparator, the 4th current comparator, the first delivery outlet,
Second delivery outlet, the 3rd delivery outlet, the 4th delivery outlet, the first electronic switch, the second electronic switch, the 3rd electronic switch and the 4th
Electronic switch, the output end of current diverter respectively with the first current comparator, the second current comparator, the 3rd current comparator
Connected with the input of the 4th current comparator, the output end of the first current comparator is exported by the first electronic switch and first
Mouth connection, the output end of the second current comparator are connected by the second electronic switch with the second delivery outlet, the 3rd current comparator
Output end be connected by the 3rd electronic switch with the 3rd delivery outlet, the output end of the 4th current comparator passes through the 4th electronic cutting
Pass is connected with the 4th delivery outlet；
The rated current of power supply unit is I, and the output current of the first delivery outlet is I1, the output current of the second delivery outlet
For I2, the output current of the 3rd delivery outlet is I3, the output current of the 4th delivery outlet is I4；Correspondingly, first current comparator
A reference value is I, and a reference value of the second comparator is I-I1, a reference value of the 3rd comparator is I-I1-I2, the base of the 4th comparator
Quasi- value is I-I1-I2-I3。
Further, the injection device uses electric injection device, the injecting method of the electric injection device include with
Lower step：
Step 1: the culture dish containing cell is positioned on locating platform first, while sticking pin and injection needle are divided
Do not tilt down and be fixed on left adjustable plate and right adjustable plate, simultaneous camera detects the image feelings of inverted microscope in real time
Condition, and signal is passed into computer；
Step 2: according to the feedback information of video camera, computer export signal feeding bit platform controller, locating platform control
Device control locating platform processed is all around moved in the horizontal direction so that culture dish is correspondingly arranged with inverted microscope；
It is as follows that the video camera carries out rectangle partitioning algorithm specific method in a scanning area to region of variation：
(1) image transmitting terminal obtains the resolution ratio of screen first, obtains 0~C of scope of column scan and the scope 0 of row scanning
~R；
(2) data of current frame image conservation zone are saved in previous frame image buffering area by transmitting terminal；Intercept and capture current screen
Curtain bitmap data is simultaneously stored in current frame image buffering area；
(3) it is (0,0) that transmitting terminal initializes change rectangular area top left co-ordinate and bottom right angular coordinate first, scanning next time
Starting point coordinate is (0,0), and row is unchanged to be identified as true, updates the scope of column scan and the scope of row scanning；
(4) judge whether to be expert in scanning range, do not exist, jump to (10)；
(5) judge whether in the range of column scan, do not exist, jump to (8)；Use in the range of column scan and directly compare every row
Current sampling point is detected compared with method；Value is different, and the unchanged mark of row is arranged into false first, then judge whether be
The first change sampled point detected, is the top left co-ordinate using sample point coordinate as change rectangular area, is not first
Individual change sampled point, relatively and maximum is taken to be sat as the new rectangle lower right corner on the coordinate of the coordinate in the rectangle lower right corner and the point
Mark, then judge whether the sampled point is first change sampled point of one's own profession, it is just by the ordinate of the sampled point with rectangle upper left
The ordinate at angle is compared and takes the top left co-ordinate of minimum value more new change rectangular area；It is worth identical, it is necessary to judge to go nothing
Change is identified whether as false, if false, starting point that record coordinate scans as next time detects it is that last row is adopted
Sampling point, the starting point using last row sample point coordinate as scanning next time, is jumped to (7)；
(6) row coordinate is moved to right N row, jumps to step 5 and detect next sampled point；
(7) one's own profession detection is finished, and the next sweep starting point of the next sweep starting point coordinate of one's own profession and lastrow record is sat
Mark compares, and takes maximum to add 1 as new next sweep starting point coordinate, line number, jump to step 4 and from the beginning opened from next line
Beginning is from left to right detected；
(8) judge that row is unchanged and identify whether that for true and change rectangular area top left co-ordinate be not (0,0), be not
True, line number add 1, jump to (4)；It is true, then shows that full line without different pixels, has obtained the rectangular area of a change
Block；Obtained change rectangular area block upper left corner ordinate be moved to the left N row, lower right corner ordinate move right N arrange with comprising
Image boundary information；
(9) the change rectangular area coordinate and corresponding next sweep starting point coordinate, judgement that record detects work as forefront
The scope of scanning whether 0~C and row scanning scope whether 0~R, be, set mark show the change rectangle that current detection goes out
Area identification is to detect for the first time, and then line number adds 1 to jump to the rectangle that (4) detect next change since next line
Region unit；Until detecting the scope beyond row scanning；
(10) after this detection, next sweep starting point all in this detection is handled, calculates next time
The set of scanning range；Whether the ordinate for first checking for this first next sweep starting point detected is adopted than last row
The ordinate of sampling point is small, is not, the region detection is completed, and detects the ordinate of next next sweep starting point；It is, with for the first time
The abscissa in the change rectangular area upper left corner detected is abscissa, is scanned related next time with currently changing rectangular area
The ordinate of point coordinates is ordinate, generates the top left co-ordinate of a next scanning range；With the change detected for the first time
The abscissa in the rectangular area lower right corner is abscissa, and a next scanning range is generated by ordinate of the maximum number of column C of screen
Bottom right angular coordinate；Then handle second next sweep starting point, until next sweep starting point all in this detection all by
Untill processing；
(11) scanning area all in next scanning range set is detected, is primarily based in next scanning range set the
The scope of the width and height of one scanning area, raw row scan and column scan, repeat (3) to (10) and detect first scanning
The rectangular area block changed in region, second scanning area is then handled, is swept until all in next scanning range set
Retouch region all be detected untill；
(12) (10) are repeated and arrives (11), the change rectangular area block of scanning range next time is obtained, until all next times
The ordinate of sweep starting point is more than or equal to the ordinate of last row sampled point, and whole screen detection finishes；
(13) the not overlapping rectangles region for the area minimum that all two field pictures change relative to previous frame image has been obtained
Set, check the rectangular area in the set, two rectangle its upper left corner ordinates are identical with lower right corner ordinate, and one
The lower right corner abscissa of rectangle is adjacent with another rectangle upper left corner abscissa, merges into a rectangle, and then recompression is concurrent
View data that the set of rectangular area is included and respective coordinates are sent to client；
(14) image-receptive end will be based on each rectangular region image data after the data decompression of reception and respective coordinates are whole
It is bonded in previous frame image and shows；
(15) (2) are repeated every the T seconds and arrives (14), according to the difference of application scenarios and the requirement of bandwidth, interval time T is done
Adjustment；
Step 3: auto-focusing is carried out to the cell in culture dish by inverted microscope, target identification finds cell；
Step 4: computer export signal gives adjustable plate controller, adjustable plate controller drives left adjustable plate operation so that
Sticking pin and cells contacting, while start linear electrical platform and drive syringe movement to produce negative pressure so that sticking pin is held carefully
Born of the same parents；
The signal r that the adjustable plate controller receiveskIt is expressed as：
In formula, A is signal amplitude, is unknown constant in a burst frame；foFor carrier wave frequency deviation, in a burst frame
For unknown constant；TsFor sampling period, foTsFor normalized carrier frequency shift；anFor QPSK modulation datas；θ0For skew,
It is unknown constant in one burst frame；G sends pulse with receiving the product of matched filter impulse function；nkFor multiple Gauss white noise
Sound, obey N (0, σ2) distribution；Timing Complete Synchronization during ε=0, otherwise timing is not synchronous；K is time sequence number, and N is over-sampling times
Number；rkThere is 10dB dynamic range；The demodulation method main task of the short preamble burst signal of low signal-to-noise ratio is from rkMiddle recovery
Go out to send data；
The computer is to signal time-frequency domain matrixPre-processed, specific bag
Include following two step：
The first step is rightLow energy is carried out to pre-process, i.e., in each sampling instant p,
WillValue of the amplitude less than thresholding ε is set to 0, and is obtained
Thresholding ε setting can determine according to the average energy of reception signal；
Second step, the time-frequency numeric field data of p moment (p=0,1,2 ... P-1) non-zero is found out, used
Represent, whereinRepresent the response of p moment time-frequencyCorresponding frequency indices, right when non-zero
The normalization pretreatment of these non-zeros, obtains pretreated vectorial b (p, q)=[b1(p,q),b2(p,q),…,bM(p,
q)]T, wherein
Step 5: the fixed injection needle of aft ramp controller regulation right adjustable plate so that injection needle connects cytoproximal
Cell membrane, now the piezoelectric ceramics power supply give the piezoelectric ceramic drive signal, piezoelectric ceramics produces vibration, realizes cell
Rupture of membranes, piezoelectric ceramics power supply is closed afterwards；
Step 6: computer export signal gives injection pump controller, injection pump controller control injection pump operation, pass through note
Penetrate pin and tissue is injected into cell desired position, complete the microinjection of cell.
Further, the adding method thereof of the medicine addition box comprises the following steps：
Step 1: receive dosing instruction；
Step 2: being instructed according to the dosing, the scarce medicine information that need to carry out feeding operations is searched by display device, if
Lookup result demonstrates the need for adding a certain or a variety of medicines, then performs next step；If lookup result shows to appoint without addition
Which kind of similar drug, then terminate dosing flow, and user need not carry out feeding operations；
Step 3: confirm the scarce medicine information of the first medicine that need to be added；
Step 4: according to the scarce medicine information of the first medicine, the first medicine is added in manipulator, and by addition
Medicine information is input in database control centre；
Step 5: the first medicine that manipulator will receive, is transported to set in advance with placing to be added first
The first dosing region corresponding to kind medicine；
Step 6: the Quantity of drugs that manipulator lacks according to the first dosing tank corresponding with the first dosing area, by the first
Medicine is added to the first dosing tank；
Step 7: the first dosing tank by the quantity of the first medicine received, feeds back to database control centre, data
Scarce medicine information in storehouse control centre more new system；
Step 8: judging whether manipulator also needs to move to other dosing areas adds the first medicine, if it is, machine
Tool hands movement adds the first medicine to other dosing areas, if it is not required, then manipulator returns to initial loading position.
Changed using speed, try to achieve corresponding OD change, and then extrapolate NADH wear rate, further extrapolated
The amount of malic dehydrogenase.MDH Lysis buffer working solutions are prepared first：MDH Lysis buffer and PMSF are taken out, it is extensive
Again to room temperature, MDH Lysis buffer：PMSF is mixed by a certain percentage, is mixed, i.e., with using, is not easy to be long placed in, otherwise white enzyme
Inhibitor PMSF efficiency can decline.Secondly NADH working solutions are prepared：NADH is taken out, recovers to room temperature, is accurately dissolved in
DdH2O, mix, 4 DEG C of pre- cold standbies.MDH is loaded afterwards, according to the testing sample 0.025MDH of MDH Lysis buffer 0.025
Assay buffer 1.8 set control tube, measure pipe, and solution should sequentially add in sequence, and pay attention to avoiding producing bubble.
If the malate dehydrogenase activity in sample is too high, it is possible to reduce is measured again after amount of samples or appropriate dilution, sample
Detection parallel pipe is preferably set.Then MDH is determined：NADH working solutions are added, immediately with spectrophotometer (1cm optical paths ratio
Color cup) measure absorbance (being designated as A0) and timing simultaneously, an absorbance is determined every 30s, wherein remembering to absorbance during 1min
For A1, its change is recorded.Leagene suggests detecting immediately after adding NADH working solutions, and loading time is more short better, its reactive group
In 1-3min, reaction thereafter tends towards stability for this.Finally according to this formula MDH (U/mlmin)=Δ A/ (0.01 × t ×
0.025) calculated, in formula：Δ A=A0-A1 (if it is necessary, the absorbance change amount for compareing initial 1min can be subtracted again)
0.01=NADH oxidations absorbance changes 0.01 per minute are that 1 enzyme activity unit t=1=detection times (min) 0.025=is treated
Survey sample volume (ml).In tissue sample MDH (U/gmin)=Δ A/ (0.01 × t × W) formula：Δ A=A0-A1 is (if any must
Will, the absorbance change amount for compare initial 1min can be subtracted again) 0.01=NADH per minute oxidation absorbance changes 0.01 are 1
Enzyme activity unit.T=1=detection times (min) W=testing samples fresh weight or dry weight (g).Tissue or plant crude enzyme liquid pick-up rate
(ml)=supernatant volume (ml)/tissue or plant quality × 100%.
It is described above to be only the preferred embodiments of the present invention, any formal limitation not is made to the present invention,
Every technical spirit according to the present invention belongs to any simple modification made for any of the above embodiments, equivalent variations and modification
In the range of technical solution of the present invention.
Claims (5)
1. a kind of malic dehydrogenase diagnostic kit, it is characterised in that the malic dehydrogenase diagnostic kit is provided with
The first casing and the second casing mutually pasted；The middle part riveting of first casing has rotating shaft, is set with and turns in the rotating shaft
Disk, some detection boxes are provided with the outside of the rotating disk；Medicine box is provided with the inside of the detection box；
Multiple injection devices are installed with second casing, detection cell, the detection are welded with the top of first casing
The upper end in pond is set with medicine addition box.
2. malic dehydrogenase diagnostic kit as claimed in claim 1, it is characterised in that the inside of the detection box is filling
There is hemostasis disinfection agent of chlorine dioxide solution, in the disinfection agent of chlorine dioxide solution that stops blooding, having comprising 0.1~50u/ml is local
The hemostasis biology enzyme of quick-acting haemostatic powder effect, 0.01~1% chlorine dioxide with the concentration that can reach disinfection effect
The Sodium azide of disinfectant and 1~100mg/ml.
3. malic dehydrogenase diagnostic kit as claimed in claim 1, it is characterised in that the rear end of the rotating disk is provided with
Drive device, supply unit is connected with the drive device, the supply unit includes：It is rechargeable battery, current diverter, more
Individual current comparator and multiple delivery outlets；
The input of current diverter is connected with rechargeable battery, the output end of current diverter and the input of each current comparator
Connection, the output end of each current comparator are respectively connecting to corresponding delivery outlet；
Also include multiple electronic switches, each electronic switch be separately positioned on corresponding between current comparator and delivery outlet, both ends
It is connected respectively with current comparator and delivery outlet；The quantity of current comparator, delivery outlet and electronic switch is four, respectively
For：First current comparator, the second current comparator, the 3rd current comparator, the 4th current comparator, the first delivery outlet,
Two delivery outlets, the 3rd delivery outlet, the 4th delivery outlet, the first electronic switch, the second electronic switch, the 3rd electronic switch and the 4th electricity
Sub switch, the output end of current diverter respectively with the first current comparator, the second current comparator, the 3rd current comparator and
The input connection of 4th current comparator, the output end of the first current comparator pass through the first electronic switch and the first delivery outlet
Connection, the output end of the second current comparator are connected by the second electronic switch with the second delivery outlet, the 3rd current comparator
Output end is connected by the 3rd electronic switch with the 3rd delivery outlet, and the output end of the 4th current comparator passes through the 4th electronic switch
It is connected with the 4th delivery outlet；
The rated current of power supply unit is I, and the output current of the first delivery outlet is I1, the output current of the second delivery outlet is I2,
The output current of 3rd delivery outlet is I3, the output current of the 4th delivery outlet is I4；Correspondingly, the benchmark of the first current comparator
It is worth for I, a reference value of the second comparator is I-I1, a reference value of the 3rd comparator is I-I1-I2, a reference value of the 4th comparator
For I-I1-I2-I3。
4. malic dehydrogenase diagnostic kit as claimed in claim 1, it is characterised in that the injection device is using electronic
Injection device, the injecting method of the electric injection device comprise the following steps：
Step 1: the culture dish containing cell is positioned on locating platform first, at the same by sticking pin and injection needle respectively to
Left adjustable plate is fixed in lower inclination and on right adjustable plate, simultaneous camera detects the image situation of inverted microscope in real time, and
Signal is passed into computer；
Step 2: according to the feedback information of video camera, computer export signal feeding bit platform controller, locating platform controller
Control locating platform is all around moved in the horizontal direction so that culture dish is correspondingly arranged with inverted microscope；
It is as follows that the video camera carries out rectangle partitioning algorithm specific method in a scanning area to region of variation：
(1) image transmitting terminal obtains the resolution ratio of screen first, obtains 0~C of scope of column scan and 0~R of scope of row scanning；
(2) data of current frame image conservation zone are saved in previous frame image buffering area by transmitting terminal；Intercept and capture current screen position
Diagram data is simultaneously stored in current frame image buffering area；
(3) it is (0,0) that transmitting terminal initializes change rectangular area top left co-ordinate and bottom right angular coordinate first, next sweep starting point
Coordinate is (0,0), and row is unchanged to be identified as true, updates the scope of column scan and the scope of row scanning；
(4) judge whether to be expert in scanning range, do not exist, jump to (10)；
(5) judge whether in the range of column scan, do not exist, jump to (8)；Used in the range of column scan every row direct comparison method
Current sampling point is detected；Value is different, and the unchanged mark of row is arranged into false first, then judges whether it is detection
The first change sampled point arrived, is the top left co-ordinate using sample point coordinate as change rectangular area, is not first change
Change sampled point, the coordinate of the coordinate in the rectangle lower right corner and the point relatively and taken into maximum as new rectangle bottom right angular coordinate,
Judge whether the sampled point is first change sampled point of one's own profession, is just by the ordinate of the sampled point with the rectangle upper left corner again
Ordinate is compared and takes the top left co-ordinate of minimum value more new change rectangular area；It is worth identical, it is necessary to judge to go unchanged
Identify whether as false, if false, starting point of the record coordinate as scanning next time, detect it is that last row samples
Point, the starting point using last row sample point coordinate as scanning next time, is jumped to (7)；
(6) row coordinate is moved to right N row, jumps to step 5 and detect next sampled point；
(7) one's own profession detection finishes, by the next sweep starting point coordinate of one's own profession and the next sweep starting point coordinate ratio of lastrow record
Compared with, and take maximum to add 1 as new next sweep starting point coordinate, line number, jump to step 4 from next line from the beginning from
Left-to-right detection；
(8) judge that row is unchanged and identify whether that for true and change rectangular area top left co-ordinate be not (0,0), be not true,
Line number adds 1, jumps to (4)；It is true, then shows that full line without different pixels, has obtained the rectangular area block of a change；
To change rectangular area block upper left corner ordinate be moved to the left N row, the ordinate N that moves right in the lower right corner is arranged with comprising image side
Boundary's information；
(9) the change rectangular area coordinate and corresponding next sweep starting point coordinate that record detects, judge current column scan
Scope whether 0~C and row scanning scope whether 0~R, be, set mark show the change rectangular area that current detection goes out
Mark is to detect for the first time, and then line number adds 1 to jump to the rectangular area that (4) detect next change since next line
Block；Until detecting the scope beyond row scanning；
(10) after this detection, next sweep starting point all in this detection is handled, calculates next scanning
The set of scope；The ordinate of this first next sweep starting point detected is first checked for whether than last row sampled point
Ordinate it is small, be not that the region detection is completed, detect the ordinate of next next sweep starting point；It is, to detect for the first time
The abscissa in the change rectangular area upper left corner gone out is abscissa, is sat with currently changing the related next sweep starting point in rectangular area
Target ordinate is ordinate, generates the top left co-ordinate of a next scanning range；With the change rectangle detected for the first time
The abscissa in the region lower right corner is abscissa, and the right side of a next scanning range is generated using the maximum number of column C of screen as ordinate
Lower angular coordinate；Then second next sweep starting point is handled, until next sweep starting point all in this detection is all processed
Untill；
(11) scanning area all in next scanning range set is detected, is primarily based in next scanning range set first
The scope of the width and height of scanning area, raw row scan and column scan, repeats (3) to (10) and detects first scanning area
The rectangular area block of middle change, second scanning area is then handled, until scanning area all in next scanning range set
Untill domain is all detected；
(12) (10) are repeated and arrives (11), obtain the change rectangular area block of scanning range next time, until all scanning next time
The ordinate of starting point is more than or equal to the ordinate of last row sampled point, and whole screen detection finishes；
(13) collection in the not overlapping rectangles region for the area minimum that all two field pictures change relative to previous frame image has been obtained
Close, check the rectangular area in the set, two rectangle its upper left corner ordinates are identical with lower right corner ordinate, and a rectangle
Lower right corner abscissa it is adjacent with another rectangle upper left corner abscissa, merge into a rectangle, then recompress and send square
The view data and respective coordinates that the set in shape region is included are to client；
(14) image-receptive end will be based on each rectangular region image data after the data decompression of reception and respective coordinates are integrated into
In previous frame image and show；
(15) (2) are repeated every the T seconds and arrives (14), according to the difference of application scenarios and the requirement of bandwidth, interval time T is adjusted
It is whole；
Step 3: auto-focusing is carried out to the cell in culture dish by inverted microscope, target identification finds cell；
Step 4: computer export signal gives adjustable plate controller, adjustable plate controller drives left adjustable plate operation so that sticking
Pin and cells contacting, while start linear electrical platform and drive syringe movement to produce negative pressure so that sticking pin holds cell；
The signal r that the adjustable plate controller receiveskIt is expressed as：
<mrow>
<msub>
<mi>r</mi>
<mi>k</mi>
</msub>
<mo>=</mo>
<mi>A</mi>
<munderover>
<mo>&Sigma;</mo>
<mrow>
<mi>n</mi>
<mo>=</mo>
<mo>-</mo>
<mi>&infin;</mi>
</mrow>
<mrow>
<mo>+</mo>
<mi>&infin;</mi>
</mrow>
</munderover>
<msub>
<mi>a</mi>
<mi>n</mi>
</msub>
<mi>g</mi>
<mrow>
<mo>(</mo>
<mfrac>
<mrow>
<msub>
<mi>kT</mi>
<mi>s</mi>
</msub>
</mrow>
<mi>N</mi>
</mfrac>
<mo>-</mo>
<msub>
<mi>nT</mi>
<mi>s</mi>
</msub>
<mo>-</mo>
<msub>
<mi>&epsiv;T</mi>
<mi>s</mi>
</msub>
<mo>)</mo>
</mrow>
<mi>exp</mi>
<mo>&lsqb;</mo>
<mi>j</mi>
<mrow>
<mo>(</mo>
<mn>2</mn>
<msub>
<mi>&pi;f</mi>
<mi>o</mi>
</msub>
<mfrac>
<mrow>
<msub>
<mi>kT</mi>
<mi>s</mi>
</msub>
</mrow>
<mi>N</mi>
</mfrac>
<mo>+</mo>
<msub>
<mi>&theta;</mi>
<mn>0</mn>
</msub>
<mo>)</mo>
</mrow>
<mo>&rsqb;</mo>
<mo>+</mo>
<msub>
<mi>n</mi>
<mi>k</mi>
</msub>
<mo>;</mo>
</mrow>
In formula, A is signal amplitude, is unknown constant in a burst frame；foIt is not in a burst frame for carrier wave frequency deviation
Know constant；TsFor sampling period, foTsFor normalized carrier frequency shift；anFor QPSK modulation datas；θ0For skew, at one
It is unknown constant in burst frame；G sends pulse with receiving the product of matched filter impulse function；nkFor white complex gaussian noise,
Obey N (0, σ2) distribution；Timing Complete Synchronization during ε=0, otherwise timing is not synchronous；K is time sequence number, and N is over-sampling multiple；rk
There is 10dB dynamic range；The demodulation method main task of the short preamble burst signal of low signal-to-noise ratio is from rkMiddle recovery is set out
Send data；
The computer is to signal time-frequency domain matrixPre-processed, specifically included as follows
Two steps：
The first step is rightLow energy is carried out to pre-process, i.e., will in each sampling instant pValue of the amplitude less than thresholding ε is set to 0, and is obtained
Thresholding ε setting can determine according to the average energy of reception signal；
Second step, the time-frequency numeric field data of p moment (p=0,1,2 ... P-1) non-zero is found out, used
Represent, whereinRepresent the response of p moment time-frequencyCorresponding frequency indices, right when non-zero
The normalization pretreatment of these non-zeros, obtains pretreated vectorial b (p, q)=[b1(p,q),b2(p,q),…,bM(p,
q)]T, wherein
Step 5: the fixed injection needle of aft ramp controller regulation right adjustable plate so that injection needle connects cytoproximal cell
Film, now the piezoelectric ceramics power supply give the piezoelectric ceramic drive signal, piezoelectric ceramics produces vibration, realizes that cell is broken
Film, piezoelectric ceramics power supply is closed afterwards；
Step 6: computer export signal gives injection pump controller, injection pump controller control injection pump operation, pass through injection needle
Tissue is injected into cell desired position, completes the microinjection of cell.
5. malic dehydrogenase diagnostic kit as claimed in claim 1, it is characterised in that the dosing of the medicine addition box
Method comprises the following steps：
Step 1: receive dosing instruction；
Step 2: being instructed according to the dosing, the scarce medicine information that need to carry out feeding operations is searched by display device, if searched
As a result demonstrate the need for adding a certain or a variety of medicines, then perform next step；If lookup result shows that any kind need not be added
Similar drug, then terminate dosing flow, and user need not carry out feeding operations；
Step 3: confirm the scarce medicine information of the first medicine that need to be added；
Step 4: according to the scarce medicine information of the first medicine, the first medicine is added in manipulator, and by the medicine of addition
Information is input in database control centre；
Step 5: the first medicine that manipulator will receive, is transported to set in advance with placing the first medicine to be added
The first dosing region corresponding to product；
Step 6: the Quantity of drugs that manipulator lacks according to the first dosing tank corresponding with the first dosing area, by the first medicine
It is added to the first dosing tank；
Step 7: the first dosing tank by the quantity of the first medicine received, feeds back to database control centre, database control
Scarce medicine information in the more new system of center processed；
Step 8: judging whether manipulator also needs to move to other dosing areas adds the first medicine, if it is, manipulator
Move to other dosing areas and add the first medicine, if it is not required, then manipulator returns to initial loading position.
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|---|---|---|---|
| CN201711137382.2A CN107841526A (en) | 2017-11-16 | 2017-11-16 | A kind of malic dehydrogenase diagnostic kit |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711137382.2A CN107841526A (en) | 2017-11-16 | 2017-11-16 | A kind of malic dehydrogenase diagnostic kit |
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ID=61679543
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| CN201711137382.2A Pending CN107841526A (en) | 2017-11-16 | 2017-11-16 | A kind of malic dehydrogenase diagnostic kit |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108753802A (en) * | 2018-05-22 | 2018-11-06 | 昆明理工大学 | One malate dehydrogenase gene CIMDH1 and its recombinant expression carrier |
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