CN209946167U - Special perforated plate for mammal oocyte and early stage blastocyst immunofluorescence - Google Patents

Abstract

A special porous plate for immunofluorescence of mammalian oocyte and early blastocyst belongs to the field of cell biology experimental equipment. The special porous plate for mammal oocyte and early stage blastocyst immunofluorescence comprises at least two groups of top plates and bottom plates, wherein the top plates are movably inserted into the bottom plates, the thickness of each top plate is larger than that of each bottom plate, each top plate comprises at least one non-penetrating operation hole and an insertion part, the operation holes are arranged on the upper portions of the top plates and are in a shape of a round step or a thread with a reduced aperture, and the top plates between each group are spliced through the insertion parts. The porous plate can simplify the transfer process of the oral suction glass tube of the oocyte and the blastocyst in the immunofluorescence staining process, improve the immunofluorescence efficiency, disperse the mammalian oocyte or the blastocyst in the operation hole, facilitate the operator to transfer or observe the mammalian oocyte or the blastocyst by using the oral suction glass tube, and effectively prevent the mammalian oocyte or the blastocyst from being lost.

Description

Special perforated plate for mammal oocyte and early stage blastocyst immunofluorescence

Technical Field

The utility model belongs to cell biology experimental device field, concretely relates to special perforated plate of mammal oocyte and early stage blastocyst immunofluorescence.

Background

Reproductive medicine research is one of the important research fields of medical research, and germ cells (primordial germ cells and mature sperms and ova) are the research objects at the core. In particular, women have limited ovarian reserve and continue to decline with age (while men have tens of millions of newborn sperm per day). More and more unfavorable internal and external environments and gene mutations causing female ovum abnormalities are identified. The utilization of oocytes of mice and other model animals as a model for researching the mechanism of oocyte maturation and blastocyst development after fertilization provides important theoretical reference for the reproductive health of human females. One of the important experimental techniques in these studies is the immunofluorescence of oocytes at various stages of the cell cycle (meiosis) to determine the cellular localization pattern of the protein of interest. The whole process of immunofluorescence requires 6 different operations, and each two steps are washed for 3 times, so that an operator needs to frequently replace a cell culture dish to perform oocyte transfer operation. Furthermore, in the case of the 2 nd penetration, the triton-containing liquid is very diffusible and must be handled in a small well with walls and a transparent bottom (e.g. a 4-well dish from Nunc). However, the walls of these wells are very tall and perpendicular to the bottom of the well, so the aspiration glass needle used by the operator to transfer the oocyte is difficult to reach the cell at the edge of the bottom of the well and the glass needle is liable to touch the upper edge of the wall of the well and break. In addition, the subsequent fixation step also tends to disperse the eggs in the bottom of the dish, making collection difficult. In summary, there is no good device for achieving a "one-stop" immunofluorescence procedure for a long time.

SUMMERY OF THE UTILITY MODEL

The technical problem to be solved is as follows: to the technical problem, the utility model provides a special perforated plate of mammal oocyte and early stage blastocyst immunofluorescence can simplify the mouth of immunofluorescence dyeing in-process oocyte and blastocyst and inhale glass tube transfer process, improves immunofluorescence efficiency to make mammal oocyte or blastocyst disperse in the handle hole, the operator of being convenient for shifts or observes mammal oocyte or blastocyst with mouth inhale glass tube, and prevents effectively that mammal oocyte or blastocyst from losing.

The technical scheme is as follows: the utility model provides a special perforated plate of mammal oocyte and early stage blastocyst immunofluorescence, special perforated plate of mammal oocyte and early stage blastocyst immunofluorescence includes at least two sets of roof and bottom plate, the roof activity is pegged graft in the bottom plate, and the roof thickness is greater than the bottom plate thickness, and the roof includes at least one operating hole and the grafting part that does not run through, the operating hole is located roof upper portion, and the operating hole is radius step form or the screw thread form that the aperture reduces, and the roof between every group passes through the grafting part concatenation.

Preferably, the plug-in components are a protruding part and a plug-in port, the protruding part comprises a horizontal protruding part and a vertical protruding part, the plug-in port comprises a horizontal plug-in port and a vertical plug-in port, the horizontal protruding part and the horizontal plug-in port are respectively arranged on the edge of one side of the top plate, the vertical protruding part and the vertical plug-in port are respectively arranged on the edge of the other side of the top plate, when the top plates are transversely spliced, the horizontal plug-in port of one top plate is inserted into the horizontal protruding part of the other top plate, and when the top plates are longitudinally spliced, the vertical plug-in port of one top plate is.

Preferably, the top of the bottom plate is provided with a groove, and the bottom of the top plate is inserted into the groove at the top of the bottom plate.

Preferably, the top plate and the bottom plate are both transparent acrylic plates.

Preferably, the mammal oocyte and early stage blastocyst immunofluorescence special multi-well plate comprises six groups of top plates and bottom plates, each group of top plates comprises 4 non-penetrating operation holes and an insertion part, and the operation holes in each group of top plates are in the shape of round steps with reduced hole diameters or in the shape of threads alternately.

Preferably, when the operation hole is in the shape of a rounded step with a reduced aperture, an included angle between the side wall of the operation hole and the vertical direction is 45 °.

Has the advantages that: 1. this novel perforated plate designs the whole required handle hole of immunofluorescence into 4X 6 form concatenation on a perforated plate, can simplify the mouth of immunofluorescence dyeing in-process oocyte and blastocyst and inhale the glass tube transfer process, improves immunofluorescence efficiency, reduces the loss rate of whole in-process oocyte and blastocyst.

2. The novel porous plate comprises at least two groups of top plates and bottom plates, wherein the top plates are movably inserted into the bottom plates, the top plates of each group are connected through insertion parts, and the detachable connection mode is more convenient for cleaning and disinfection of the subsequent porous plate;

3. the shape of this novel handle hole is the radius step form or the heliciform that the aperture reduces, makes mammal oocyte or blastocyst disperse in the handle hole, and the operator of being convenient for shifts or observes mammal oocyte or blastocyst with mouth suction glass pipe to prevent effectively that mammal oocyte or blastocyst from losing, avoid among the prior art mouth suction glass pipe 5 be difficult to reach the cell of hole bottom edge and easily touch the situation that the pore wall upper limb was broken, simultaneously because the constraint of hole also can save reagent (especially expensive one resist and the quantity of two antibodies).

Drawings

FIG. 1 is a top view of an operation hole of embodiment 1 in a rounded step shape;

FIG. 2 is a sectional view of the mammalian oocyte and early stage blastocyst immunofluorescence specific multi-well plate A-A of example 1;

FIG. 3 is a top view of the operating hole of example 2 in a spiral shape with a reduced diameter;

FIG. 4 is a sectional view of the mammalian oocyte and early stage blastocyst immunofluorescence specific multi-well plate A-A of example 2.

The numerical designations in the drawings represent the following: 1. a top plate; 2. a base plate; 3. an operation hole; 4. a mammalian oocyte or blastocyst; 5. the mouth sucks the glass tube.

Detailed Description

The invention is further described with reference to the accompanying drawings and specific embodiments.

Example 1

A special multi-hole plate for immunofluorescence of mammalian oocytes and early blastula is shown in figures 1 and 2 and comprises six groups of top plates 1 and bottom plates 2. The top plate 1 and the bottom plate 2 are both transparent acrylic plates. The top plate 1 is movably inserted into the bottom plate 2. The thickness of the top plate 1 is greater than that of the bottom plate 2. The top plate 1 comprises 4 non-through operating holes 3 and plug-in parts. The operation hole 3 is arranged at the upper part of the top plate 1, the operation hole 3 is in the shape of a circular step with a reduced aperture, the included angle between the side wall of the operation hole and the vertical direction is 45 degrees, mammalian oocytes or blastocysts 4 can be dispersed in the operation hole 3, an operator can transfer or observe the mammalian oocytes or blastocysts 4 conveniently by using the mouth suction glass tube 5, the mammalian oocytes or blastocysts 4 are effectively prevented from being lost, the situation that the mouth suction glass tube 5 in the prior art cannot reach the cells at the edge of the bottom of the hole easily and can be broken by touching the upper edge of the hole wall is avoided, and meanwhile, reagents (especially the dosage of expensive primary antibody and secondary antibody) can be saved due to the constraint of the operation hole 3. The top plates 1 between each group are spliced through the splicing parts. Roof 1 activity is pegged graft in bottom plate 2, connects through insertion parts between every group roof, and the cleaning and the disinfection of follow-up perforated plate are convenient for more to detachable connected mode.

Example 2

A special perforated plate for immunofluorescence of mammalian oocytes and early blastula is disclosed, and referring to fig. 3 and 4, the special perforated plate for immunofluorescence of mammalian oocytes and early blastula comprises 6 groups of top plates 1 and bottom plates 2, and the top plates 1 and the bottom plates 2 are transparent acrylic plates. The top plate 1 is movably inserted into the bottom plate 2, a groove is arranged at the top of the bottom plate 2, and the bottom of the top plate 1 is inserted into the groove at the top of the bottom plate 2. The thickness of the top plate 1 is greater than that of the bottom plate 2. The top plate 1 includes 4 non-penetrating operation holes 3 and insertion members, and the arrangement order of the operation holes on the top plate 1 is 2 (rows) × 2 (columns). The plug-in components are protruding portions and plug-in ports, the protruding portions comprise horizontal protruding portions and vertical protruding portions, the plug-in ports comprise horizontal plug-in ports and vertical plug-in ports, the edges of one side of the top plate 1 are respectively arranged on the horizontal protruding portions and the horizontal plug-in ports, the edges of the other side of the top plate 1 are respectively arranged on the vertical protruding portions and the vertical plug-in ports, when the top plates are transversely spliced, the horizontal plug-in ports of one top plate are inserted into the horizontal protruding portions of the other top plate, and when the top plates are longitudinally spliced, the vertical plug-in ports of one top plate are inserted into the vertical protruding. The operation hole 3 is arranged at the upper part of the top plate 1, the operation hole 3 is in a thread shape with a reduced aperture, so that mammalian oocytes or blastocysts are dispersed in the operation hole, an operator can transfer or observe the mammalian oocytes or blastocysts conveniently by using the mouth-sucking glass tube 5, the mammalian oocytes or blastocysts are effectively prevented from being lost, the situation that in the prior art, the mouth-sucking glass tube 5 cannot reach cells at the edge of the bottom of the hole easily and can be broken by touching the upper edge of the hole wall easily is avoided, and reagents (especially the dosage of expensive primary antibody and secondary antibody) can be saved due to the restriction of the hole. Roof 1 between every group is through the concatenation of grafting part, and the roof activity is pegged graft in the bottom plate, connects through grafting part between every group roof, and the cleaning and the disinfection of follow-up perforated plate of being convenient for more of detachable connected mode. The splicing mode among 6 top plates is 2 (rows) × 3 (columns), and the sequence of the operation holes of the special multi-hole plate for mammal oocyte and early blastocyst immunofluorescence is 4 (rows) × 6 (columns). The multi-hole plate designs the operation holes required in the whole immunofluorescence process into a 4 x 6 format and is spliced on the multi-hole plate, so that the process of transferring the oral suction glass tubes of the oocytes and the blastocysts in the immunofluorescence staining process can be simplified, the immunofluorescence efficiency is improved, and the loss rate of the oocytes and the blastocysts in the whole process is reduced.

The specific use method of the mammal oocyte and early blastocyst immunofluorescence special porous plate is as follows: when the operator performs immunofluorescence on the oocyte or blastocyst, a sample (the mammalian oocyte or blastocyst 4) is moved to the multi-well plate through the oral suction glass tube 5, and then the sample is sequentially rinsed 3 times → 1 time of penetration → 3 times of rinsing → 1 time of fixation → 3 times of rinsing → 1 time of primary anti-incubation → 3 times of rinsing → 1 time of secondary antibody incubation → DNA staining → 1 time of rinsing, and 22 holes are needed in total, so that all operation steps of immunofluorescence are sufficiently completed on all the holes.

Example 3

The difference from example 2 is that the top plate 1 is 3 mm thick, the bottom plate 2 is 1.5 mm thick, 12 cm long and 8 cm wide. The top surface of the operation hole 3 is larger than the bottom surface, the thickness of the operation hole 3 is 2 mm, the diameter of the top surface is 1.2 cm, and the diameter of the bottom surface is 0.8 cm. The shape of the operation hole 3 in each group of top plates 1 is a round step shape with reduced aperture or a thread shape which is alternated, so that the identification degree is increased. When the operation hole is in the shape of a round step with a reduced aperture, the included angle between the side wall of the operation hole and the vertical direction is 45 degrees, so that the situation that the mouth suction glass tube 5 in the prior art is difficult to reach the cells at the edge of the bottom of the hole and is easy to touch the upper edge of the hole wall to break can be avoided.

Claims (6)

1. The utility model provides a special perforated plate of mammal oocyte and early stage blastocyst immunofluorescence, its characterized in that, special perforated plate of mammal oocyte and early stage blastocyst immunofluorescence includes at least two sets of roof (1) and bottom plate (2), roof (1) activity is pegged graft in bottom plate (2), and the thickness of roof (1) is greater than the thickness of bottom plate (2), and roof (1) includes at least one handling hole (3) and the grafting part that does not run through, roof (1) upper portion is located in handling hole (3), and handling hole (3) are radius step form or the screw thread form that the aperture reduces, and roof (1) between every group are through the grafting part concatenation.

2. The special multi-hole plate for immunofluorescence of oocytes and early stage blastocysts of mammals according to claim 1, wherein the insertion parts are a protrusion part and an insertion port, the protrusion part comprises a horizontal protrusion part and a vertical protrusion part, the insertion port comprises a horizontal insertion port and a vertical insertion port, the horizontal protrusion part and the horizontal insertion port are respectively arranged on the edge of one side of the top plate (1), the vertical protrusion part and the vertical insertion port are respectively arranged on the edge of the other side of the top plate (1), when the two top plates are transversely spliced, the horizontal insertion port of one top plate is inserted into the horizontal protrusion part of the other top plate, and when the two top plates are longitudinally spliced, the vertical insertion port of one top plate is inserted into the vertical protrusion part of the other top plate.

3. The special multi-well plate for immunofluorescence of mammalian oocyte and early stage blastocyst according to claim 1, wherein the top of the bottom plate (2) is provided with a groove, and the bottom of the top plate (1) is inserted into the groove at the top of the bottom plate (2).

4. The mammalian oocyte and early stage blastocyst immunofluorescence special multi-well plate according to claim 1, wherein, the top plate (1) and the bottom plate (2) are both transparent acrylic plates.

5. The special perforated plate for immunofluorescence of mammalian oocytes and early blasts according to claim 1, which comprises six groups of top plates (1) and bottom plates (2), wherein each group of top plates (1) comprises 4 non-penetrating operation holes (3) and an insertion part, and the operation holes (3) in each group of top plates (1) are in the shape of round steps with reduced pore diameters or are in an alternating thread shape.

6. The special multiwell plate for immunofluorescence of mammalian oocyte and early stage blastocyst according to claim 1, wherein when the operation hole (3) is a rounded step shape with reduced aperture, the angle between the side wall of the operation hole (3) and the vertical direction is 45 °.

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