CN216209185U - Cell slide is preserved and immunochemistry dyeing apparatus - Google Patents

Abstract

The utility model discloses a cell slide preservation and immunochemical staining device, belonging to the field of sample staining devices for testing. The cell slide glass comprises a climbing glass slide, a glass slide clamp and a staining jar, wherein the climbing glass slide comprises a marking area and a cell climbing glass bearing area which is uniformly provided with cylindrical climbing glass bearing grooves for placing cell climbing glasses; the glass slide clamp comprises a clamping plate and a handle connected with the clamping plate; one or more immersion tanks are arranged in the dyeing tank, the side wall of the dyeing tank is provided with a long hole for the handle to penetrate out, and the bottom of the dyeing tank is provided with a water outlet hole and leveling support legs which penetrate through the immersion tanks. The utility model has the advantages that the experiment operation is convenient and fast by arranging the matched staining jar, the glass slide and the glass slide clamp, cells can be directly stained and marked in the staining jar at the same time, the consistency is good, and the product preservation is convenient by fixing paraffin; the utility model has high integration and strong compatibility, can save reagents and reduce test cost.

Description

Cell slide is preserved and immunochemistry dyeing apparatus

Technical Field

The utility model belongs to the field of sample dyeing devices for testing, and particularly relates to a cell slide preservation and immunochemical dyeing device.

Background

Cells are important tools for molecular biology research, and immunocytochemistry is an important molecular biology technology for detecting the expression level of genes, molecules, proteins and the like of cells. At present, if the cellular immunochemical staining is carried out, cells need to be revived and cultured, and are inoculated on a glass slide in a culture plate, and a staining experiment can be carried out in the culture plate when the cells grow to a certain density. Although this experiment is widely used in work, the following problems exist:

(1) the experiment was repeated: after the researchers complete one-time cell staining, related gene and protein indexes are further screened according to experimental results and recent progress at home and abroad to carry out cell staining experiments, cells need to be cultured again before each time of staining, and a large amount of working time and experimental reagents are wasted.

(2) Cell lysis: although the cells growing on the slide can be fixed by reagents such as ethanol or acetone and then stored in a refrigerator at the temperature of-20 ℃, the cells can not be stored for a long time after being fixed, and the cells have a dissolution phenomenon along with the prolonging of the storage time, so that the subsequent test can not be carried out.

(3) The experimental consistency is poor: when immunochemical staining is carried out on the cell glass slide in the culture plate, the steps of staining, decoloring, anti-blue, developing and the like of various chemical substances cannot be carried out simultaneously in batches, so that the staining condition difference of the cell glass slide of each hole is large, and the experimental result is influenced.

SUMMERY OF THE UTILITY MODEL

The utility model is provided for overcoming the defects in the prior art, and aims to provide a cell slide preservation and immunochemical staining device.

The utility model is realized by the following technical scheme:

a cell slide preservation and immunochemical staining device comprises a slide glass, a glass clip and a staining cylinder, wherein the slide glass comprises a marking area and a cell slide bearing area which is uniformly provided with a cylindrical slide bearing groove for placing cell slides; the glass slide clamp comprises a clamping plate and a handle connected with the clamping plate; in the dyeing tank, one or more immersion tanks are arranged in the dyeing tank, the side wall of the dyeing tank is provided with a long hole for the handle to penetrate out, and the bottom of the dyeing tank is provided with a water outlet hole and leveling support legs which penetrate through the immersion tanks.

In the technical scheme, the cross section of the immersion tank is matched with the bottom cross section of the glass slide clamp on which the slide glass is placed.

In the technical scheme, the bottom of the immersion tank is inclined downwards from the side edge to the center, the water outlet hole is formed in the center of the immersion tank, and a water plug is arranged in the water outlet hole.

In the technical scheme, the dyeing device further comprises a limiting lifting device, the limiting lifting device comprises a rack arranged in the dyeing cylinder and a limiting mechanism, and the limiting mechanism comprises a gear arranged in the dyeing cylinder and meshed with the rack, a connecting pipe which is connected with the gear and penetrates through the long hole, and a knob connected with the other end of the connecting pipe.

In the technical scheme, the handle is provided with the limiting snap ring.

In the above technical solution, the total length of the slide glass is 120mm, the width is 26mm, and the thickness is 4mm, wherein the length of the marking region is 15mm, and the length of the cell slide bearing region is 105 mm.

In the technical scheme, on the same slide glass, the number of the slide bearing grooves is eight, the diameter of the slide bearing grooves is 10mm, and the distance between every two slide bearing grooves is 2 mm.

In the technical scheme, on the same slide glass, the number of the slide bearing grooves is five, the diameter of the slide bearing grooves is 16mm, and the distance between every two slide bearing grooves is 4 mm.

In the above technical scheme, the depth of the creeper blade bearing groove is 2 mm.

In the technical scheme, capital letter marks are arranged right above the climbing sheet bearing grooves.

The utility model has the beneficial effects that:

(1) the operation is convenient and fast, and the consistency is good: the experimenter can directly put the cell-overgrowing slide in the slide glass bearing groove, so that the cells can be directly dyed and marked in the dyeing cylinder, and the consistency is good.

(2) The sample is convenient to preserve: after the cell slide is fixed on the glass slide by paraffin, the cell sample can be preserved at normal temperature for a long time without being put into a refrigerator with the temperature of 20 ℃ below zero, thereby saving energy and avoiding the cell autolysis phenomenon.

(3) Integration is high, compatibility is strong: the cell climbing film fixing and dyeing of 8-hole or 5-hole cell climbing film can be realized by one climbing film glass slide, the batch processing is convenient, and the climbing film glass slide with the specification can be directly placed in an immunohistochemical frame applied in a laboratory at present, so that the compatibility is strong.

(4) Reagent saving: for rare and expensive reagents such as the first antibody, the slide glass has great advantages, after the slide is added into the slide bearing groove, 50ul of the reagent is added to cover the whole slide, and reagent waste is greatly avoided.

Drawings

FIG. 1 is a schematic view of the external structure of the present invention;

FIG. 2 is a schematic cross-sectional view taken along line A-A of the present invention;

FIG. 3 is a schematic structural view of the limit lifting device of the present invention;

FIG. 4 is a schematic external view of an eight well slide of the present invention;

FIG. 5 is a schematic external view of a five-well slide according to the present invention.

Wherein:

1. a slide, 101, a marking zone,

102. a slide bearing groove 103, a cell slide bearing area,

2. dyeing vat, 201, water block,

202. an immersion tank, 203, a long hole,

204. water outlet holes, 205, leveling feet,

206. a handle, 3. a clamping plate,

301. a handle, 302. an L-shaped inserting piece,

3021. a horizontal plug board, 3022, a vertical plug board,

303. clip, 304, limit snap ring

4. A limit lifting device 401, a rack,

402. a gear, 403, a connecting pipe,

404. a knob.

Detailed Description

In order to make the technical solutions of the present invention better understood by those skilled in the art, the technical solutions of the present invention are further described below by way of specific embodiments with reference to the drawings of the specification.

Example 1:

a cell slide preservation and immunochemical staining device comprises a slide glass 1, a slide glass clamp and a staining cylinder 2, wherein the slide glass 1 comprises a marking area 101 and a cell slide bearing area 103 which is uniformly provided with a cylindrical slide bearing groove 102 for placing cell slides; the glass slide clamp comprises a clamping plate 3 and a handle 301 connected with the clamping plate 3; the slide glass 1 can be arranged on the clamping plate 3, and the handle 301 can be held to move the slide glass 1; one or more immersion tanks 202 are arranged inside the dyeing cylinder 2, the side wall of the dyeing cylinder 2 is provided with a long hole 203 for the handle to penetrate out, and the bottom of the dyeing cylinder 2 is provided with a water outlet 204 penetrating through the immersion tanks 202 and leveling support feet 205.

The holding plate 3 is used for holding one end of the marking area 101, the handle 301 penetrates through the long hole 203, and the experimental operation of the movement of the slide glass 1 can be completed by holding the handle 301. The operation of putting and lifting the slide glass 1 from the immersion tank 202 can be completed by moving the handle 301 up and down; the rotation of the slide glass 1 can be completed by rotating the handle 301, and the redundant liquid on the slide glass 1 is drained.

Example 2:

on the basis of embodiment 1, the number of the immersion tanks 202 is four, and the immersion tanks are arranged at the bottom of the dyeing cylinder 2 side by side. The cross section of the immersion tank 202 is matched with the bottom cross section of the slide holder with the slide glass 1, so that the reagent can be saved in the immersion process, and the slide glass 1 can be immersed by a small amount of reagent. The bottom of the immersion tank 202 is inclined downwards from the side edge to the center, the water outlet 204 is arranged in the center of the immersion tank 202, namely the water outlet 204 is arranged at the lowest position of the bottom of the immersion tank 202, the water plug 201 is arranged in the water outlet 204, and the handle 206 is arranged at the lower end of the water plug 201, so that waste liquid of the immersion tank 202 can be conveniently discharged at any time without dumping the dyeing tank.

Example 3:

on the basis of the above embodiment, the grip block 3 includes L-shaped inserting plate 302 and clamping piece 303, L-shaped inserting plate 302 includes horizontal inserting plate 3021 of the chinese character 'shan' shape and vertical inserting plate 3022 connected with horizontal inserting plate 3021, clamping piece 303 is connected on vertical inserting plate 3022 and is parallel to horizontal inserting plate 3021, and handle 301 is L-shaped and is connected with vertical inserting plate 3022 upside.

Meanwhile, the utility model also comprises a limiting lifting device 4, wherein the limiting lifting device 4 comprises a rack 401 arranged in the dyeing cylinder 2 and a limiting mechanism, the limiting mechanism comprises a gear 402 which is arranged in the dyeing cylinder 2 and is meshed with the rack 401, a connecting pipe 403 which is connected with the gear 402 and penetrates through the long hole 203, and a knob 404 which is connected with the other end of the connecting pipe 403, the handle penetrates through the gear 402, the connecting pipe 403 and the knob 404 in sequence, and a limiting snap ring 304 can be further arranged on the handle to help the handle to maintain the position relative to the limiting mechanism.

The knob 404 is screwed, and the limit lifting device drives the handle 301 to move up and down in the long hole 203, so that after the handle 301 is loosened, the clamping plate 3 can also maintain the relative position with the immersion tank 202, and the operation of long-time immersion or draining is convenient.

Example 4:

on the basis of the above examples, the slide glass 1 has a length of 120mm, a width of 26mm and a thickness of 4 mm. The length of the marking area 101 is 15mm, the width thereof is 26mm, and the length of the cell slide bearing area 103 is 105mm, and the width thereof is 26 mm. The climbing piece glass slide 1 comprises two specifications, one is an eight-hole glass slide, namely the number of the climbing piece bearing grooves on the glass slide is eight, the diameter of the climbing piece bearing groove 102 is 10mm, the climbing piece bearing groove is used for placing climbing pieces with the diameter of 8mm, and the climbing piece bearing grooves 102 are separated by 2 mm; the other type is a five-hole glass slide, namely, the number of the slide bearing grooves is five, the diameter of the slide bearing groove 102 is 16mm, the slide bearing groove is used for placing 14 mm-diameter slide, and the distance between every two slide bearing grooves 102 is 4 mm. The arrangement is convenient for batch processing, and the slide glass 1 with the specification can be directly placed in an immunohistochemical frame applied in a laboratory at present, so that the compatibility is strong.

The mark area 101 is an area printed by 15mm × 26mm by using a thermal transfer printing technology, so that characters can be written conveniently.

The cell slide bearing area 103 marks each slide bearing groove 102 by capital letters, transversely marks each hole site (8mm is A-H, 14mm is A-E) in sequence, and marks the position right above the slide bearing groove 102 to clearly determine the grouping and positioning of each slide bearing groove 102.

In order to carry out a high-throughput test on the slide glass, numerical control machine tool holes are formed in the positions of each slide bearing groove 102, the holes are respectively 10mm and 16mm for cell slides with the cell thicknesses of 8mm and 14mm, and the depths are both 2 mm.

And (3) completely spraying paraffin on the slotted slide glass 1, printing and marking the position of the capital letters A-H, A-E after wax spraying, etching the exposed capital letters by hydrofluoric acid, and dewaxing and washing to obtain the clearly marked slide glass 1.

Example 5:

the use method of the utility model comprises the following steps:

when in use, the prepared cell slide is directly placed in the slide bearing groove 102 of the slide glass 1, and then the next immunocytochemistry experiment can be carried out.

If the immunohistochemistry experiment is not carried out temporarily, the slide can be placed in the slide bearing groove 102, then the immunohistochemical dehydration step is carried out, finally paraffin embedding is directly carried out in the slide bearing groove 102, the cell slide paraffin specimen is prepared for long-term storage, and when needed, the staining observation can be carried out after dewaxing hydration according to the immunohistochemical process.

The use steps are further detailed below:

firstly, directly carrying out immunocytochemistry staining experimental steps on cells in the slide bearing groove 102

1. Fixing: cells on slide 1 containing the cell slide were washed 3 times with 1 × PBS for 3min each time. Fixation with freshly prepared 4% paraformaldehyde for 15min at room temperature.

2. Rinsing: the fixative was removed by rinsing 3 times with 1 XPBS for 5min each time.

3. Permeabilization: 0.1% Triton X-100 was added to 1 XPBS and incubated for 10 minutes.

4. Rinsing: the residual liquid was removed and rinsed 3 times with 1 × PBS for 5min each.

5. And (3) sealing: residual liquid was removed and 5% BSA or serum was added to the sections. Ensure that the serum and secondary antibody are derived from the same species. Then incubated at room temperature for 20 min.

6. Incubating the primary antibody: the residual liquid was removed and the tissue sites were covered with primary antibody at the appropriate dilution ratio and incubated overnight at 4 ℃.

7. Rewarming: the sections were allowed to stand from 4 ℃ to room temperature for about 20min without immediate washing of the sections.

8. Rinsing: the residual liquid was removed and rinsed 3 times with 1 × PBS for 5min each.

9. Incubating fluorescently conjugated secondary antibodies: the residual liquid was removed, the tissue sites were covered with secondary antibody at the appropriate dilution ratio and incubated at 4 ℃ for 15min in the dark.

10. Rinsing: the residual liquid was removed and rinsed 3 times with 1 × PBS for 5min each.

11. Color development: adding DAB color developing agent, and standing at room temperature for 3 min.

12. Rinsing: the residual liquid was removed and rinsed 3 times with 1 × PBS for 5min each.

13. Hematoxylin staining: adding the threo lignin staining solution, and keeping the temperature for 5 min.

14. Rinsing: rinsing with tap water for 5 min.

15. Differentiation: the cells were differentiated with 1% HCl for 30s and washed with tap water for 5 min.

16. Returning blue: returning the blue to 0.2% ammonia water for 30s, and washing with tap water for 5 min.

17. Dewatering and sealing: dehydrating with 50%, 75%, 90%, 100% gradient alcohol for 3min each, air drying, and sealing with neutral gum.

Second, cell slide paraffin fixing method

1. Fixing: the slide was transferred to slide-carrying tank 102, and the cells on slide 1 containing the cell slide were washed 3 times with 1 × PBS for 3min each time. Fixation with freshly prepared 4% paraformaldehyde for 15min at room temperature.

2. And (3) dehydrating: the fixed cells were dehydrated for 3min by gradient alcohol of 50%, 75%, 90%, 100%.

3. Wax dipping: the creep plate bearing groove 102 containing creep plates is directly soaked in wax for embedding, and can be stored for a long time at room temperature after being embedded.

Thirdly, paraffin-embedded cell slide dewaxing and hydration experiment method:

1. baking: the slide bearing groove 102 containing the cell slide is baked in a 60 ℃ oven for 20 min.

2. Dewaxing: the slide bearing tank 102 was immersed in xylene and dewaxed for 30 min.

3. Hydration: hydrating with 100%, 90%, 75%, 50% gradient alcohol for 3min each.

4. Rinsing: rinse 3 times with 1 × PBS for 5min each time.

5. After the above treatment, immunocytochemical staining can be performed by cell slide (see experiment step one).

The utility model provides a cell slide preservation and immunochemical staining device, which achieves the following effects by arranging a staining cylinder, a glass slide and a glass slide clamp which are matched with each other:

(1) the operation is convenient and fast, and the consistency is good: the experimenter can directly put the cell-overgrowing slide in the slide glass bearing groove, so that the cells can be directly dyed and marked in the dyeing cylinder, and the consistency is good.

(2) The sample is convenient to preserve: after the cell slide is fixed on the glass slide by paraffin, the cell sample can be preserved at normal temperature for a long time without being put into a refrigerator with the temperature of 20 ℃ below zero, thereby saving energy and avoiding the cell autolysis phenomenon.

(3) Integration is high, compatibility is strong: the cell slide with 6 holes or 5 holes can be fixed and dyed by one slide glass, so that the batch treatment is convenient, and the slide glass with the specification can be directly placed in an immunohistochemical frame applied in a laboratory at present, and has strong compatibility.

(4) Reagent saving: for rare and expensive reagents such as the first antibody, the slide glass has great advantages, after the slide is added into the slide bearing groove, 50ul of the reagent is added to cover the whole slide, and reagent waste is greatly avoided.

It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.

In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on those shown in the drawings, and are used only for convenience in describing the present invention and for simplicity in description, and do not indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and thus, are not to be construed as limiting the present invention. Furthermore, the terms "first", "second", etc. are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first," "second," etc. may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless otherwise specified.

In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meaning of the above terms in the present invention can be understood by those of ordinary skill in the art through specific situations.

The applicant declares that the above description is only a specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and it should be understood by those skilled in the art that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are within the scope and disclosure of the present invention.

Claims (10)

1. The utility model provides a cell slide is preserved and immunochemistry dyeing apparatus which characterized in that: the cell slide glass comprises a slide glass (1), a slide glass clamp and a staining jar (2), wherein the slide glass (1) comprises a marking area (101) and a cell slide bearing area (103) which is uniformly provided with a cylindrical slide bearing groove (102) for placing cell slides; the glass slide clamp comprises a clamping plate (3) and a handle (301) connected with the clamping plate (3); the inside of dyeing jar (2) is equipped with one or more immersion tank (202), the lateral wall of dyeing jar (2) is equipped with slot hole (203) that supply handle (301) to wear out, dyeing jar (2) bottom is equipped with apopore (204) and leveling stabilizer blade (205) that link up immersion tank (202).

2. The device for cell slide preservation and immunochemical staining according to claim 1, wherein: the cross section of the immersion tank (202) is matched with the bottom cross section of the glass slide clamp on which the slide glass (1) is placed.

3. The device for cell slide preservation and immunochemical staining according to claim 2, wherein: the bottom of the immersion tank (202) is inclined downwards from the side edge to the center, the water outlet hole (204) is formed in the center of the immersion tank (202), and a water plug (201) is arranged in the water outlet hole (204).

4. The device for cell slide preservation and immunochemical staining according to claim 1, wherein: the dyeing device is characterized by further comprising a limiting lifting device (4), wherein the limiting lifting device (4) comprises a rack (401) arranged in the dyeing cylinder (2) and a limiting mechanism, and the limiting mechanism comprises a gear (402) which is arranged in the dyeing cylinder (2) and is meshed with the rack (401), a connecting pipe (403) which is connected with the gear (402) and penetrates through the long hole (203), and a knob (404) which is connected with the other end of the connecting pipe (403).

5. The device for cell slide preservation and immunochemical staining according to claim 4, wherein: the handle (301) is provided with a limiting snap ring (304).

6. The device for cell slide preservation and immunochemical staining according to claim 1, wherein: the total length of the slide glass (1) is 120mm, the width is 26mm, the thickness is 4mm, the length of the marking area (101) is 15mm, and the length of the cell slide bearing area (103) is 105 mm.

7. The device for cell slide preservation and immunochemical staining according to claim 6, wherein: on the same slide glass (1) of climbing film, the quantity of climbing film bearing groove (102) is eight, and its diameter is 10mm, and each climbing film bearing groove (102) is separated by 2 mm.

8. The device for cell slide preservation and immunochemical staining according to claim 6, wherein: on the same slide glass (1) of climbing film, the quantity of climbing film bearing groove (102) is five, its diameter is 16mm, every climbing film bearing groove (102) is 4mm apart.

9. The cell-slide preserving and immunochemical staining apparatus as claimed in claim 7 or 8, wherein: the depth of the climbing piece bearing groove (102) is 2 mm.

10. The device for cell slide preservation and immunochemical staining according to claim 9, wherein: capital letter marks are arranged right above each climbing sheet bearing groove (102).

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