CN115181656A - Preparation method of drug-mixed micro-slab by using drip plate - Google Patents
Preparation method of drug-mixed micro-slab by using drip plate Download PDFInfo
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- CN115181656A CN115181656A CN202210934971.8A CN202210934971A CN115181656A CN 115181656 A CN115181656 A CN 115181656A CN 202210934971 A CN202210934971 A CN 202210934971A CN 115181656 A CN115181656 A CN 115181656A
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- C12M23/00—Constructional details, e.g. recesses, hinges
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- C12M23/00—Constructional details, e.g. recesses, hinges
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- C12M23/38—Caps; Covers; Plugs; Pouring means
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- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/48—Holding appliances; Racks; Supports
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
- C12M41/22—Heat exchange systems, e.g. heat jackets or outer envelopes in contact with the bioreactor walls
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
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- 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
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/18—Testing for antimicrobial activity of a material
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/2806—Means for preparing replicas of specimens, e.g. for microscopal analysis
Abstract
The invention relates to microbiology and plant pathology technology, in particular to an agar micro-plate containing medicament prepared by mixing a tiny amount of medicament sample liquid with water agar under the working condition of a micro-plate sample mixing table of a 70 ℃ temperature system by adopting a drip plate. The main components of the microplate sample mixing table comprise a temperature control body A, a sample mixing drip plate preheating component B, a drug sample preheating component C, a gun head preheating component D and a glass slide preheating ceramic tile plate component E. The preparation method of the drug microplate mainly comprises the following steps: 1) preparing a medicine sample, 2) preparing a mixed sample by using water agar, 3) processing a gun head, 4) starting a sample mixing table to enter a working state, 5) preheating the sample, 6) preheating a glass slide, 7) injecting sample liquid, 8) injecting water agar, 9) uniformly mixing, and 10) unfolding the mixed sample into a medicine microplate. The invention has the advantages that: 1) The drip plate can be repeatedly sterilized for use; 2) The total amount of drug samples required is minimal; 3) The prepared drug microplate is convenient for subsequent biological determination and microscopic observation.
Description
Technical Field
The invention relates to microbiology and plant pathology technology, in particular to a preparation method of a drug micro-slab mixed by a drip plate.
Technical Field
Plant diseases caused by harmful microorganisms can cause huge economic loss and ecological damage, and the main means for preventing and controlling the plant diseases is still pesticide prevention and control at present and in a considerable period in the future. Exploring and exploring biological source medicine is an important direction for pesticide control. When a sample with inhibitory activity is searched and screened from a plurality of samples of microbial sources or plant sources, if the inhibitory activity can be detected and identified by little metabolic fluid (or interstitial fluid) in the primary screening stage, the screening efficiency can be improved, and the screening range can be enlarged. In the research process of biological-source medicines, the inhibition effect of metabolites generated in the culture process of beneficial microorganisms on harmful microorganisms is explored, the current analysis and detection technology often needs to terminate the culture of some culture bottles so as to filter the culture products and prepare enough detection samples, and the technicians in the field most hope to directly extract trace samples from the culture bottles for biological assay without terminating the culture process. In the case where microorganisms (or plants) are found to produce inhibitory metabolites (or plant tissue fluids), but the inhibitory components are still unknown, isolation of the active is generally required, and in the process of isolating the active, it is often necessary to perform a follow-up assay of the biological activity on a small number of isolated samples. Obviously, the research and development work of the biogenic pesticide needs technical support of detection of trace or small amount of drug samples.
The conventional technical method for detecting the inhibitory activity of a drug on harmful microorganisms in the current field is mainly a plate culture determination method, and the technical key point of the method is that the drug is mixed with water agar to prepare a drug-containing culture plate, then the harmful microorganisms are implanted on the drug-containing plate, the drug-containing culture plate is cultured under proper conditions, and the inhibitory activity of the drug is measured and calculated by observing and comparing the growth amount of the harmful microorganisms on the plate. The basic technology of the method is the preparation of a drug-containing culture plate, and the conventional technology of the current preparation of the drug-containing culture plate is to mix a drug sample and water agar in a triangular flask and pour the mixture in a conventional culture dish to prepare the drug-containing culture plate. Although the preparation method of the drug-containing culture plate is simple and mature, the preparation method has an obvious characteristic that the sample amount of the drug to be detected is required to be more than dozens of milliliters, which is difficult to match with the detection technology of trace or small amount of sample expected in the field, so the preparation method is an important defect of the current drug-containing culture plate preparation technology.
Disclosure of Invention
The invention aims to provide a method for preparing a drug-containing microplate by mixing a tiny amount of drug sample liquid with water agar by using a dropping plate.
The invention designs and adopts the technical conditions of the micro-plate sample mixing table to solve the technical problems, and the technical scheme for solving the problems is as follows:
1. a micro-plate sample mixing table is designed, and comprises a temperature control body A, a sample mixing drip plate preheating component B, a medicine sample preheating component C, a gun head preheating component D and a glass slide preheating ceramic tile plate component E. The sample mixing table is adaptive to the superclean bench.
The temperature control body A mainly comprises a basin-shaped container A1, a water body A2, a water body electric heating temperature control member A3 and a water body water surface cover plate A4. The electric heating temperature control component A3 of the water body can automatically heat the water body and stabilize the temperature of the water body to be in a working state of 70 ℃, and the cover plate A4 of the water surface of the water body can prevent the water body from being quickly evaporated and cooled.
The drip plate preheating component B for sample mixing mainly comprises a drip plate B1 and a drip plate cushion frame B2, wherein the drip plate B1 is half thick and is immersed in the water body A2. The medicine sample preheating component C mainly comprises a sample tube C1 and a sample tube rack C2, and the tube body of the sample tube C1 is directly immersed into the water body A2. The gun head preheating component D mainly comprises a gun head box D1 and a gun head box clamping frame D2, and the box body of the gun head box (D1) is immersed in the water body A2. The glass slide preheating ceramic tile plate component E mainly comprises a ceramic tile plate E1 and a ceramic tile plate bracket E2, and part of the ceramic tile plate E1 is immersed in a water body A2.
2. The preparation method for preparing the drug-containing microplate by using the designed microplate sample mixing platform mainly comprises the following steps of putting the sample mixing platform into an ultra-clean workbench, using a dropping plate as a mixing device of drugs and agar, using a ceramic tile plate as a preheating plate of a glass slide, and preparing the drug microplate in a technical system with a water body at 70 ℃, wherein the preparation method comprises the following steps:
1) Preparation of drug samples: a small amount of drug sample obtained during drug development work was transferred into the sterilized sample tube C1.
2) Preparing a mixed sample by using water agar: the mixture ratio of water agar for mixing samples is as follows: agar 40g and water 1000ml, heating to melt, and maintaining in water bath at 90 deg.C or above.
3) Gun head processing: using a conventional tip with the order of 200 μ l, a small section of the tip is cut off, and then the tip with the tip cut off is loaded into a tip box, sterilized and dried in a conventional manner, and the tip box is fixed on a tip box holder D2.
4) Starting a sample mixing table to enter a working state: the method comprises the steps of firstly, normally starting an operating superclean bench, then placing a sample mixing table on a working table surface inside the superclean bench, taking a sterilized dropping plate, horizontally placing the dropping plate on a dropping plate pad frame B2 frame, then switching on a power supply of the sample mixing table, electrically heating and regulating the working state of the water body A2 to 70 ℃, and keeping the operation till the dropping plate B1, a gun head in a gun head box D1 and a glass slide preheating ceramic tile plate E1 are in a working state balanced with the temperature of the water body at 70 ℃.
5) Inserting the medicine sample tube C1 in the step 1) into the tube hole of the sample tube rack C2 floating on the water surface of the water body A2, and preheating in the water body A2.
6) And (3) taking the sterilized glass slide, and horizontally placing the sterilized glass slide on the surface of the glass slide preheating tile plate E1 for preheating.
7) Sleeving the gun head in the working state in the step 4) on a liquid transfer gun, sucking 100 mu l of sample liquid from the preheated sample tube in the step 5), and injecting the sample liquid into a recess of the drip plate B1 in the working state in the step 4) to form a sample liquid drop at the bottom of the recess; .
8) Covering the operating gun head of the step 4) with a liquid transfer gun, sucking 30 mu l of the mixed sample water agar of the step 2), injecting into the sample liquid drop formed in the step 7), keeping the liquid transfer gun head in the liquid drop, repeating the liquid suction and liquid discharge actions, washing the residual agar in the lower gun head, and obtaining the mixed sample liquid of the sample liquid and the water agar.
9) Sleeving a pipette tip with the working state in the step 4) on a pipette tip with the adjusted pipette tip volume of 150 mu l, inserting the pipette tip into the sample mixing liquid obtained in the step 8), and performing repeated liquid suction and liquid discharge actions to fully and uniformly mix the sample liquid in the sample mixing liquid with the water agar.
10 Completely sucking the uniform sample mixing liquid obtained in the step 9) into a gun head, transferring to the preheated glass slide obtained in the step 6), discharging the sample mixing liquid of the liquid-transfering gun onto the surface of the glass slide at different positions of the glass slide in a segmented point form, stirring and flattening the discharged liquid points by using the gun head to enable each liquid point to form a thin agar micro-block, transferring the glass slide into a sterile container with normal temperature and moisture preservation, and condensing the thin agar micro-block to obtain the drug micro-plate.
THE ADVANTAGES OF THE PRESENT INVENTION
1) The drip plate can be repeatedly sterilized for use.
2) The invention can mix 0.1ml of medicine sample liquid into a plurality of medicine micro-flat plates, and needs a little total amount of medicine samples.
3) The drug-containing microplate prepared based on the glass slide is convenient for subsequent biological determination and microscopic observation.
Drawings
FIG. 1 is a schematic front view of a microplate sample mixing station. In the figure, A is a temperature control body component, B is a drip plate preheating component for mixing samples, C is a medicine sample preheating component, D is a gun head preheating component, and E is a glass slide preheating ceramic tile plate component.
Fig. 2 is a schematic side view of the temperature control body a. In the figure, A1 is a basin-shaped container, A2 is a water body, A3 is a water body electric heating temperature control component, and A4 is a water surface cover plate of the water body.
Fig. 3 is a schematic side structure diagram of each parallel component in the sample mixing table and a schematic space relationship diagram between the components and the temperature control body water body. In the figure, A1 is a basin-shaped container of the temperature control body, A5 is a bottom plane line of the basin-shaped container of the temperature control body, and A6 is a horizontal line of the water body surface of the temperature control body. B is a preheating component of the drip plate for sample mixing, B1 is the drip plate, and B2 is the drip plate cushion frame. C is a preheating component of a medicine sample, C1 is a sample tube, and C2 is a sample tube rack. D is a gun head preheating component, D1 is a gun head box, and D2 is a gun head box clamping frame. E is a glass slide preheating ceramic tile plate component, E1 is a ceramic tile plate, and E2 is a ceramic tile plate bracket.
FIG. 4 is a schematic view of a pipette gun sucking a sample mixture and dropping the sample mixture on the surface of a glass slide, in which a rectangular frame is the glass slide and a dark gray circle spot is a drop of the sample mixture.
FIG. 5 is a schematic view showing a method of preparing a drug-containing microplate by mixing a drip plate according to the present invention, in which garlic tissue fluid is mixed with agar and the mixture is dripped on a common glass slide to form 5 drug-containing microplates.
Detailed Description
Under the condition of a tiny amount of a medicine sample, tiny medicine sample liquid and tiny water agar are mixed conventionally, the water agar is often not ready to be mixed with the medicine sample liquid and is solidified in a liquid transferring instrument, so that the work cannot be carried out, and obviously, under the conventional technical condition, a medicine-containing flat plate with tiny samples is difficult to prepare. The invention skillfully designs a set of micro-plate sample mixing table, establishes a proper sample mixing technical condition system, can effectively and uniformly mix trace drug sample liquid and trace high-concentration water agar, and forms a thin layer on a glass slide to be developed, so as to obtain the micro-plate containing the drug.
The main components of the microplate sample mixing table comprise a temperature control body A, a sample mixing drip plate preheating component B, a drug sample preheating component C, a gun head preheating component D and a glass slide preheating tile plate component E, as shown in FIG. 1. The whole micro-flat plate sample mixing table can be placed on the inner tabletop of the ultra-clean workbench, the related operations of the invention are implemented under the working running state of the ultra-clean workbench, and the prepared drug-containing micro-flat plate is ensured to be in a sterile state.
The sectional structure of each main member is shown in fig. 2 and 3.
The main components of the temperature control body A comprise a basin-shaped container A1, a water body A2 contained in the container, a water body electric heating temperature control component A3 and a water surface cover plate A4 of the water body, as shown in figure 2. The electric heating temperature control member A3 of the water body can automatically heat the water body and stabilize the temperature of the water body to be in a working state of 70 ℃, and the cover plate A4 on the water surface of the water body can prevent the water body from being quickly evaporated and cooled.
The droplet plate preheating member B for sample mixing mainly includes a droplet plate B1 and a droplet plate holder B2, as shown in fig. 3B. The drip plate is a ceramic drip plate widely used in a chemical laboratory, and a recess on the drip plate is used for mixing a sample and agar; the drip plate cushion frame B2 is embedded in the water of the temperature control body water body A2, and can support the drip plate B1 into a stable state; half of the thickness of the drip plate B1 is immersed into the water body A2, and the temperature of the water body A2 can be quickly balanced.
The medicine sample preheating component C mainly comprises a sample tube C1 and a sample tube rack C2, as shown in a C picture of figure 3, a tube hole is formed in the sample tube rack C2 and floats on the water surface of a water body A2 of the temperature control body, and a small centrifugal tube of 1.5ml is adopted for the sample tube C1; after the sample tube C1 is inserted into the tube hole of the tube frame C2, the sample tube body is directly immersed into the water of the water body A2, so that the sample tube and the sample liquid in the tube can be quickly balanced to the temperature of the water body A2.
The lance tip preheating member D mainly includes a lance tip cartridge D1 and a lance tip cartridge holder D2, as shown in fig. 3D. The gun head box clamping frame D2 is also embedded into the water body A2 of the temperature control body, so that a common gun head box can be clamped and stably fixed, and the box body of the gun head box D1 is maintained to be immersed into the water body A2, so that the gun head box D1 and a gun head arranged in the gun head box D1 can quickly balance the temperature of the water body A2. The cover of the gun head box D1 is completely exposed outside the water body A2, and the gun head can be taken by opening the box at will.
The slide preheating tile plate member D mainly includes a slide preheating tile plate D1 and a tile plate holder D2, as shown in D of fig. 3. The ceramic tile plate bracket D2 is immersed in the water body A2 of the temperature control body and can stably support the ceramic tile plate D1; the ceramic tile board D1 is formed by stacking 2 common floor ceramic tiles with the thickness of 1cm, the bottom ceramic tile board D1 is half thick and is buried in the water A2, the temperature of the water A2 can be quickly balanced by the ceramic tile board, and the temperature of the surface of the upper ceramic tile board can be reduced to about 43 ℃ due to heat resistance and conduction of a gap between the upper ceramic tile board and the two ceramic tile boards.
The preparation method for preparing the drug-containing microplate by using the designed microplate sample mixing table technically comprises the steps of putting the sample mixing table into an ultra-clean workbench, using a spot plate as a mixing tool of drugs and agar, using a ceramic tile plate as a preheating plate of a glass slide, and preparing the drug microplate in a technical system with water at 70 ℃, wherein the preparation method comprises the following steps:
1. preparation of drug samples: a small sample of drug taken during drug development work is transferred into the sterilized sample tube C1.
2. Preparing a mixed sample by using water agar: the mixture ratio of water agar for mixing samples is as follows: agar 40g and water 1000ml, sterilizing at high temperature, and heating to melt before use. The water agar is viscous after being melted, a large number of bubbles are easily generated when the water agar is heated and boiled, and the bubbles are difficult to fade. When the water agar is completely melted, it is maintained in a water bath at a temperature above 90 ℃ and placed in a position within reach of the operator.
3. Gun head processing: the preparation operation mainly uses 200 mul of gun head, because 200 mul of gun head has small diameter of tip mouth and high viscosity water agar can not pass through smoothly, the tip of the gun head is cut off by about 5-8mm by a cutting tool such as a blade, thereby obtaining the opening of the gun head with larger diameter and ensuring the water agar to pass through smoothly. And (4) putting the tip-cut gun heads into a gun head box in a conventional mode, sterilizing and drying, and fixing on a gun head clamping frame D2.
4. Starting a sample mixing table to enter a working state: firstly, normally starting and operating an ultra-clean workbench, then placing a micro-flat plate sample mixing table on a working table top in the ultra-clean workbench, taking a sterilized dropping plate, flatly placing the sterilized dropping plate on a dropping plate pad frame B2 frame of the sample mixing table, then switching on a power supply of the sample mixing table, electrically heating a water body A2 to a working state with the temperature of 70 ℃, and maintaining the working state to operate for more than 20 minutes, so that the temperatures of the dropping plate B1 and a gun head in a gun head box D1 and the water body A2 reach balance and are in the working state; the temperature of the surface of the glass slide preheating ceramic tile plate E1 is kept at about 43 ℃ in the working state.
5. And (3) inserting the medicine sample tube C1 in the step (1) into the tube hole of the sample tube rack C2 floating on the water surface of the water body A2, immersing the tube body of the sample tube in the water body, and preheating for more than 5 minutes.
6. And (3) taking the sterilized glass slide, placing the sterilized glass slide on the surface of the glass slide preheating tile plate E1, and preheating for more than 5 minutes.
7. And (3) sleeving the gun head in the working state in the step (4) by using a liquid transfer gun, sucking 100 mu l of sample liquid from the preheated sample tube in the step (5), and injecting the sample liquid into a cavity of the dropping plate B1 in the working state in the step (4) to form a sample liquid drop at the bottom of the cavity.
8. And (4) sleeving the pipette head in the working state in the step (4) by using a liquid transfer gun, sucking 30 mu l of the water agar for mixing the sample in the step (2), injecting the sample into the sample liquid drop formed in the step (7), keeping the pipette head in the liquid drop, repeatedly carrying out liquid suction and liquid discharge for about 5 times, washing the residual agar in the pipette head, and obtaining the sample liquid and the water agar mixed sample liquid.
9. And (3) sleeving the pipette head in the working state in the step (4) by using a pipette gun with the adjusted liquid suction amount of 150 mu l, inserting the pipette head into the sample mixing liquid obtained in the step (8), and performing repeated liquid suction and liquid discharge actions for about 5 times to fully and uniformly mix the sample liquid in the sample mixing liquid with the water agar.
10. Sucking all the uniformly mixed sample liquid obtained in the step 9 into a gun head, transferring to the preheated glass slide in the step 6, discharging the sample liquid of a liquid transfer gun onto the surface of the glass slide in a segmented dotted manner at different positions of the glass slide, as shown in fig. 4, pulling and flattening the discharged liquid points by using the gun head to enable each liquid point to form a thin agar micro-block, transferring the glass slide into a sterile container with normal temperature and moisture preservation, placing the glass slide in the sterile container, condensing the thin agar micro-block to form a drug micro-plate, and generally, injecting the sample liquid for one time on the glass slide to form 5-10 drug micro-plates.
After the preparation work of the drug micro-flat plate is finished, the power is cut off to stop the operation work of the micro-flat plate sample mixing table, and relevant components of the sample mixing table are cleaned, dried and properly stored for later use.
The design of the invention is specifically implemented in a laboratory with the normal temperature of 25 ℃. The main technology is that a spot plate is used as a sample mixing device, a ceramic tile plate of a common floor ceramic tile is used as a preheating plate of a glass slide, and the water body of a micro-plate sample mixing table is designed to be 70 ℃; preheating of sample mixing equipment, preheating of samples, preheating of gun heads and preheating of glass slides are all organically integrated into a micro-flat plate sample mixing table system. Such a thermal equilibrium technology system can avoid the mixed sample liquid in the drop plate recess to be in an excessive high temperature state and lead to rapid evaporation of water, can also avoid rapid evaporation of water of the microplate developed on the glass slide, and can prevent agar condensation in the operation process at the same time. Since the total amount (130 μ l) of the sample mixing solution is small, bubbles are easily generated in the sample mixing operations of the liquid suction and liquid discharge in step 9 of the preparation method, and the speed of the liquid suction and liquid discharge operations needs to be observed and controlled to reduce the formation of bubbles, which is beneficial to obtaining the bubble-free agar microplate.
The operation from step 7 to step 10 of the method aims at a tiny amount of sample liquid and water agar and is operated under a higher temperature condition, so the operation is coherent, and the long-time pause in the operation process is avoided, thereby not only reducing the condensation probability of the water agar in the operation process, but also reducing the evaporation and the loss of water in the sample mixing liquid. In practice, 3 pipetting guns with fixed pipetting volumes can be used to adjust and fixedly aspirate 100 μ l of drug sample solution, 30 μ l of water agar and 150 μ l of water agar for mixing the sample solution, so that the pause of adjusting the pipetting volumes in the middle of using one pipetting gun alone can be avoided.
Example 1
Taking 1ml of garlic tissue grinding extract (extraction ratio: 1g of garlic, 5ml of water) as a medicine sample solution. By using the method for preparing a micro-plate by mixing a drug sample solution with water agar according to the present invention, a micro-plate containing garlic tissue fluid was prepared, and as a result, a micro-plate containing garlic tissue fluid without bubbles was obtained, as shown in FIG. 5.
Claims (2)
1. A drug microplate sample mixing table is characterized in that main components comprise a temperature control body (A), a sample mixing drip plate preheating component (B), a drug sample preheating component (C), a gun head preheating component (D) and a glass slide preheating ceramic tile plate component (E), and the sample mixing table is adapted to be applied to an ultra-clean workbench;
the main components of the temperature control body (A) comprise a basin-shaped container (A1), a water body (A2), a water body electric heating temperature control component (A3) and a water body water surface cover plate (A4), wherein the water body electric heating temperature control component (A3) can automatically heat the water body and stabilize the working state of the water body to 70 ℃, and the water body water surface cover plate (A4) can prevent the water body from quickly evaporating and cooling;
the sample mixing drip plate preheating component (B) mainly comprises a drip plate (B1) and a drip plate cushion frame (B2), wherein the half thickness of the drip plate (B1) is immersed in a water body (A2); the medicine sample preheating component (C) mainly comprises a sample tube (C1) and a sample tube rack (C2), and the tube body of the sample tube (C1) is directly immersed into the water body (A2); the gun head preheating component (D) mainly comprises a gun head box (D1) and a gun head box clamping frame (D2), and the box body of the gun head box (D1) is immersed in the water body (A2); the glass slide preheating ceramic tile plate component (E) mainly comprises a ceramic tile plate (E1) and a ceramic tile plate bracket (E2), and part of the ceramic tile plate (E1) is immersed in a water body (A2).
2. A method of preparing a drug-containing microplate using the drug microplate mixing platform of claim 1, characterized in that the drug-containing microplate is prepared in a technical system in which the water is at 70 ℃ by placing the mixing platform on a clean bench, using a spot plate as a mixing implement for the drug and agar, and using a tile plate as a preheating plate for the slide glass, the method comprising the steps of:
1) Preparation of drug samples: transferring a small amount of drug samples obtained in drug research and development work into a sterilized sample tube (C1);
2) Preparing a mixed sample by using water agar: the mixture ratio of water agar for mixing samples is as follows: heating agar 40g and water 1000ml to melt, and maintaining in water bath at 90 deg.C above;
3) Gun head treatment: cutting a small section of a tip of a conventional gun head with the magnitude of 200 mu l, then loading the gun head with the tip cut off into a gun head box in a conventional mode, sterilizing and drying, and then fixing the gun head box on a gun head box clamping frame (D2);
4) Starting a sample mixing table to enter a working state: firstly, normally starting and operating an ultra-clean workbench, then placing a sample mixing table on a working table top in the ultra-clean workbench, then taking a sterilized dropping plate, flatly placing the dropping plate on a dropping plate pad frame (B2), then switching on a power supply of the sample mixing table, electrically heating and regulating the temperature of a water body (A2) to a working state of 70 ℃, and maintaining and operating until the dropping plate (B1), a gun head in a gun head box (D1) and a glass slide preheating tile plate (E1) are in a working state of being balanced with the temperature of the water body of 70 ℃;
5) Inserting the medicine sample tube (C1) in the step 1) into a tube hole of a sample tube frame (C2) of a floating frame on the water surface of a water body (A2), and preheating in the water body (A2);
6) Placing the sterilized glass slide on the surface of a glass slide preheating ceramic tile plate (E1) for preheating;
7) Sleeving the gun head in the working state in the step 4) by using a liquid transfer gun, sucking 100 mu l of sample liquid from the preheated sample tube in the step 5), and injecting the sample liquid into a recess of the drip plate (B1) in the working state in the step 4) to form a sample liquid drop at the bottom of the recess;
8) Sleeving a gun head in a working state in the step 4) by using a liquid transfer gun, sucking 30 mu l of the mixed sample water agar in the step 2), injecting into the sample liquid drop formed in the step 7), keeping the liquid transfer gun head in the liquid drop, repeatedly sucking and discharging liquid, washing residual agar in the lower gun head, and obtaining mixed sample liquid of the sample liquid and the water agar;
9) Sleeving a liquid transferring gun with the adjusted liquid suction amount of 150 mu l on the gun head in the working state in the step 4), inserting the gun head into the sample mixing liquid obtained in the step 8), and performing repeated liquid suction and liquid discharge actions to fully and uniformly mix the sample liquid in the sample mixing liquid with the water agar;
10 And) sucking all the uniform sample mixing liquid obtained in the step 9) into a gun head, transferring to the preheating glass slide obtained in the step 6), discharging the sample mixing liquid of the liquid transfer gun onto the surface of the glass slide at different positions of the glass slide in a segmented point form, stirring and flattening the discharged liquid points by using the gun head to enable all the liquid points to form a thin agar micro-block, transferring the glass slide into a sterile container with normal temperature and moisture preservation, and condensing the thin agar micro-block to obtain the drug micro-plate.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210934971.8A CN115181656A (en) | 2022-07-28 | 2022-07-28 | Preparation method of drug-mixed micro-slab by using drip plate |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210934971.8A CN115181656A (en) | 2022-07-28 | 2022-07-28 | Preparation method of drug-mixed micro-slab by using drip plate |
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| Publication Number | Publication Date |
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| CN202210934971.8A Pending CN115181656A (en) | 2022-07-28 | 2022-07-28 | Preparation method of drug-mixed micro-slab by using drip plate |
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