Imaging method and device for self-luminous objects on biological sample membrane

Abstract

The present invention discloses an imaging method and device for a self-luminous object on a biological sample film, wherein the imaging device comprises a shell, a photoelectric conversion element and an image correction device; the interior of the shell constitutes a darkroom space; the photoelectric conversion element is arranged in the shell; the photoelectric conversion element is used to obtain a first dark field image inside the shell when the biological sample film is not placed; after the biological sample film is attached to the surface of the photoelectric conversion element, the photoelectric conversion element is also used to obtain a second dark field image inside the shell; the image correction device is used to correct the second dark field image according to the first dark field image to obtain a target image corresponding to the self-luminous object. The present invention can effectively and more accurately obtain a target image of a self-luminous object with higher clarity, the operation process is simple, the imaging time is short, and the imaging device has the advantages of small structure, low manufacturing cost, convenient operation and easy carrying.

Description

Technical Field

[0001] The present invention relates to the technical field of sample analysis, and in particular to an imaging method and device for a self-luminous object on a biological sample film. Background Art

[0002] There are currently two types of commonly used bioluminescence detection technologies, one is using photosensitive film technology, and the other is using camera shooting technology.

[0003] Among them, the photosensitive film technology is similar to the traditional photographic film developing method. In a dark room, the self-luminous object (attached to the biological sample membrane) is tightly attached to the photosensitive film, and then the exposed film is re-washed with a developer to obtain the image corresponding to the self-luminous object; however, the imaging method has a complicated operation process and requires more steps for digital processing; the camera shooting technology is similar to the shooting technology of a mobile phone. The camera and the self-luminous object are placed in a dark room. This method can directly store digital images, but due to the long distance between the camera and the self-luminous object, the light signal acquisition rate is easily low, so it takes a long time to collect data, and there are defects such as low sensitivity. In addition, the above two imaging systems have defects such as large size, large space occupation, high cost, and difficulty in transportation. Summary of the invention

[0004] The technical problem to be solved by the present invention is that the imaging method of obtaining self-luminous objects on biological sample films in the prior art has the disadvantages of complex operation process, long imaging time, large size, large space occupation, high cost, and difficulty in transportation of the imaging system. The purpose is to provide a method and device for imaging self-luminous objects on biological sample films.

[0005] The present invention solves the above technical problems through the following technical solutions:

[0006] The present invention provides an imaging device for a self-luminous object on a biological sample film, the imaging device comprising a housing, a photoelectric conversion element and an image correction device;

[0007] The interior of the shell forms a darkroom space;

[0008] The photoelectric conversion element is disposed in the housing;

[0009] The photoelectric conversion element is used to obtain a first dark field image of the interior of the housing within a set acquisition time when no biological sample membrane is placed therein;

[0010] Wherein, the biological sample film carries a self-luminous object;

[0011] After the biological sample film is attached to the surface of the photoelectric conversion element, the photoelectric conversion element is further used to obtain a second dark field image of the interior of the shell within the set acquisition time;

[0012] The image correction device is used to perform correction processing on the second dark field image according to the first dark field image to obtain a target image corresponding to the self-luminous object.

[0013] Preferably, the image correction device performs correction processing on the second dark field image according to the first dark field image, and obtains the formula corresponding to the target image corresponding to the self-luminous object as follows:

[0014] I=I0-I d

[0015] Wherein, I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d Represents pixel data corresponding to the first dark field image.

[0016] Preferably, the imaging device further comprises a light source device, and the light source device is arranged in the housing.

[0017] Preferably, the photoelectric conversion element is also used to obtain a bright field image inside the housing within the set acquisition time after the biological sample film is not placed in the housing and the light source device in the darkroom space is turned on;

[0018] The image correction device is also used to perform correction processing on the second dark field image according to the first dark field image and the bright field image to obtain a target image corresponding to the self-luminous object.

[0019] Preferably, the photoelectric conversion element is also used to obtain a bright field image corresponding to the set acquisition time when the biological sample film is not placed in the housing, the housing is in an open state, and a uniform light field is provided by an external light source;

[0020] The image correction device is also used to perform correction processing on the second dark field image according to the first dark field image and the bright field image to obtain a target image corresponding to the self-luminous object.

[0021] Preferably, the image correction device performs correction processing on the second dark field image according to the first dark field image and the bright field image, and obtains the formula corresponding to the target image corresponding to the self-luminous object as follows:

[0022]

[0023] Wherein, I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d represents the pixel data corresponding to the first dark field image, I f Represents pixel data corresponding to the bright field image.

[0024] Preferably, the housing comprises a light shielding cover and a base;

[0025] One side of the light shielding cover is hinged to one side of the base;

[0026] When the light shielding cover and the base are closed, the interior of the shell forms the darkroom space.

[0027] Preferably, the light source device is arranged at the top position of the inner side of the light shielding cover; and / or,

[0028] The corresponding illumination duration after the light source device is turned on is 10ms-30s; and / or,

[0029] The light source device includes a plurality of LED (light emitting diode) lamp beads arranged in a dot matrix, a plurality of lamps introduced by optical fibers, a plurality of lamp tubes arranged in parallel or a plurality of plate-shaped lamps.

[0030] Preferably, the imaging device further comprises a diffuser plate;

[0031] The light diffuser is fixedly mounted on the inner side of the light shielding cover and is located directly below the light source device; and / or,

[0032] The photoelectric conversion element includes a CMOS (complementary metal oxide semiconductor) chip, a CCD (charge coupled device) chip or an amorphous silicon photoelectric conversion detector; and / or,

[0033] The biological sample membrane includes a protein membrane, an agarose gel block, an agarose gel strip, a polyacrylamide gel block or a polyacrylamide gel strip.

[0034] Preferably, the imaging device further comprises a protective film;

[0035] Both sides of the protective film are respectively attached to the biological sample film and the photoelectric conversion element.

[0036] Preferably, the thickness of the protective film is 0.01 mm-0.2 mm; and / or,

[0037] The material of the protective film is a tempered glass film or a hard plastic film.

[0038] The present invention also provides a method for imaging a self-luminous object on a biological sample film, the imaging method is implemented using the above-mentioned imaging device for a self-luminous object on a biological sample film, and the imaging method comprises:

[0039] Using the photoelectric conversion element to acquire a first dark field image of the interior of the housing within a set acquisition time;

[0040] After the biological sample film is attached to the surface of the photoelectric conversion element, the photoelectric conversion element is used to obtain a second dark field image of the interior of the housing within the set acquisition time;

[0041] Wherein, the biological sample film carries a self-luminous object;

[0042] The second dark field image is corrected according to the first dark field image to obtain a target image corresponding to the self-luminous object.

[0043] Preferably, the formula corresponding to the step of correcting the second dark field image according to the first dark field image to obtain the target image corresponding to the self-luminous object is as follows:

[0044] I=I0-I d

[0045] Wherein, I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d Represents pixel data corresponding to the first dark field image.

[0046] Preferably, the imaging device further comprises a light source device;

[0047] Before the step of correcting the second dark field image according to the first dark field image to obtain the target image corresponding to the self-luminous object, the step further includes:

[0048] The photoelectric conversion element is used to obtain a bright field image inside the housing within the set acquisition time after the biological sample film is not placed in the housing and the light source device in the darkroom space is turned on;

[0049] The step of correcting the second dark field image according to the first dark field image to obtain the target image corresponding to the self-luminous object comprises:

[0050] The second dark-field image is corrected according to the first dark-field image and the bright-field image to obtain a target image corresponding to the self-luminous object.

[0051] Preferably, before the step of correcting the second dark field image according to the first dark field image to obtain the target image corresponding to the self-luminous object, the step further includes:

[0052] The photoelectric conversion element is used to obtain a bright field image corresponding to the set acquisition time when the biological sample film is not placed in the housing, the housing is in an open state, and a uniform light field is provided by an external light source;

[0053] The step of correcting the second dark field image according to the first dark field image to obtain the target image corresponding to the self-luminous object comprises:

[0054] The second dark-field image is corrected according to the first dark-field image and the bright-field image to obtain a target image corresponding to the self-luminous object.

[0055] Preferably, the formula corresponding to the step of correcting the second dark-field image according to the first dark-field image and the bright-field image to obtain the target image corresponding to the self-luminous object is as follows:

[0056]

[0057] Wherein, I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d represents the pixel data corresponding to the first dark field image, I f Represents pixel data corresponding to the bright field image.

[0058] The positive and progressive effects of the present invention are:

[0059] The interior of the shell in the image acquisition device of the present invention constitutes a darkroom space. When no self-luminous object is placed, the photoelectric conversion element first obtains a first dark field image. After the self-luminous object is placed, the photoelectric conversion element obtains a second dark field image. The second dark field image is then corrected according to the first dark field image to obtain a target image corresponding to the self-luminous object. In addition, the light field generated by the light source device in the shell or the bright field image under the light field generated by the external light source can be combined to more effectively and accurately obtain a target image of the self-luminous object with higher clarity. The operation process is simple and the imaging time is short. The imaging device has the advantages of small structure, low manufacturing cost, convenient operation and easy carrying. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1 This is a first structural schematic diagram of the imaging device of a self-luminous object on a biological sample film according to Embodiment 1 of the present invention.

[0061] Figure 2 This is a second structural schematic diagram of the imaging device for a self-luminous object on a biological sample film according to Embodiment 1 of the present invention.

[0062] Figure 3This is a schematic diagram of the first shell structure in the imaging device of a self-luminous object on a biological sample film according to Example 1 of the present invention.

[0063] Figure 4 This is a schematic diagram of the second shell structure in the imaging device of the self-luminous object on the biological sample film according to Example 1 of the present invention.

[0064] Figure 5 This is a schematic diagram of the first dark field imaging in the imaging device of the self-luminous object on the biological sample film according to Example 1 of the present invention.

[0065] Figure 6 This is a schematic diagram of the second dark field imaging in the imaging device of the self-luminous object on the biological sample film according to Example 1 of the present invention.

[0066] Figure 7 This is a first structural schematic diagram of an imaging device for a self-luminous object on a biological sample film according to Embodiment 2 of the present invention.

[0067] Figure 8 This is a second structural schematic diagram of the imaging device for a self-luminous object on a biological sample film according to Embodiment 2 of the present invention.

[0068] Fig. 9 This is a third structural schematic diagram of the imaging device for a self-luminous object on a biological sample film according to Embodiment 2 of the present invention.

[0069] Fig.10 Schematic diagram of bright field imaging in an imaging device for a self-luminous object on a biological sample film according to Example 2 of the present invention.

[0070] Fig.11 This is a schematic diagram of the relationship between the lanes and the signal intensity in the imaging device of the self-luminous object on the biological sample film according to Example 2 of the present invention.

[0071] Fig.12 This is a schematic diagram of the second dark field imaging in the imaging device of the self-luminous object on the biological sample film according to the second embodiment of the present invention.

[0072] Fig.13 This is a schematic structural diagram of an imaging device for a self-luminous object on a biological sample film according to Embodiment 3 of the present invention.

[0073] Fig.14 This is a flow chart of a method for imaging a self-luminous object on a biological sample film according to Embodiment 4 of the present invention.

[0074] Fig.15 This is a flow chart of a method for imaging a self-luminous object on a biological sample film according to Embodiment 5 of the present invention.

[0075] Fig.16 This is a flow chart of the imaging method of a self-luminous object on a biological sample film according to Example 6 of the present invention. DETAILED DESCRIPTION

[0076] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples.

[0077] Example 1

[0078] like Figure 1 and Figure 2 As shown, the imaging device of a self-luminous object on a biological sample film of this embodiment includes a housing 1 , a photoelectric conversion element 2 and an image correction device 3 .

[0079] The interior of the housing 1 forms a darkroom space, wherein the housing 1 includes a light shielding cover 4 and a base 5. Figure 3 As shown, one side of the shading cover 4 is hinged to one side of the base 5, and at this time the shading cover 4 is in an open state.

[0080] like Figure 4 As shown, when the light shielding cover 4 and the base 5 are closed, the interior of the housing 1 forms a dark room to ensure that the biological sample membrane (in Figure 2 The weak signal corresponding to the luminous object (represented by A in the figure) can be effectively collected.

[0081] Among them, biological sample membranes include protein membranes, agarose blocks, agarose strips, polyacrylamide gel blocks, polyacrylamide gel strips, etc.

[0082] The photoelectric conversion element is arranged in the housing, wherein the photoelectric conversion element includes a CMOS chip, a CCD chip or an amorphous silicon photoelectric conversion detector.

[0083] The photoelectric conversion element is used to obtain a first dark field image (such as Figure 5 shown);

[0084] Among them, the biological sample film carries a self-luminous object.

[0085] The collection time can be set and adjusted according to actual conditions. Preferably, the collection time is set to 1 second.

[0086] Specifically, when the biological sample membrane includes a protein membrane, the protein membrane sample is separated by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel) electrophoresis technology, and then the protein is transferred to a PVDF (polyvinylidene fluoride) membrane in a buffer to obtain a biological sample membrane carrying the protein. The biological sample membrane is sequentially incubated with a target protein-specific first-level antibody and a horseradish peroxidase-coupled second-level antibody, and then developed with a chemiluminescent liquid (i.e., displayed as a self-luminous object), generating a chemical light signal in the area of ​​the target protein.

[0087] After the biological sample film is attached to the surface of the photoelectric conversion element, the photoelectric conversion element is also used to obtain a second dark field image (such as Figure 6 As shown), that is, the data acquisition time lengths corresponding to the first dark field image and the second dark field image are consistent.

[0088] The image correction device is used to perform correction processing on the second dark field image according to the first dark field image to obtain a target image corresponding to the self-luminous object.

[0089] The process of acquiring the first dark field image using the photoelectric conversion element includes:

[0090] Open the light shielding cover 4 to remove the stains and the like remaining on the stage (i.e., the photoelectric conversion element) in the imaging device;

[0091] Close the light shielding cover 4 so that the interior of the housing 1 forms a darkroom space to ensure that the photoelectric conversion element is not affected by external light signals;

[0092] Wait for a set time (such as 1s), and use the photoelectric conversion element to collect the value I of each pixel in the uniform dark field inside the shell. d , where the collection time is 1s.

[0093] The process of acquiring the second dark field image using the photoelectric conversion element includes:

[0094] Open the light shielding cover 4, and place the biological sample film carrying the self-luminous object on the stage (i.e., the photoelectric conversion element) so that the biological sample film and the photoelectric conversion element are closely attached;

[0095] Close the light shielding cover 4 so that the interior of the housing 1 forms a darkroom space to ensure that the photoelectric conversion element is not affected by external light signals;

[0096] A photoelectric conversion element is used to collect the light signal corresponding to the self-luminous object (target protein) on the biological sample film (at this time, other areas outside the target protein belong to a uniform dark field). The photoelectric conversion element collects the value I0 of each pixel point inside the shell, and the collection time is 1s.

[0097] The image correction device performs correction processing on the second dark field image according to the first dark field image, and obtains the formula corresponding to the target image corresponding to the self-luminous object as follows:

[0098] I=I0-I d

[0099] Where I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d Indicates pixel data corresponding to the first dark field image.

[0100] The size of the second dark field image is consistent with that of the first dark field image, I0-I d The difference represents the difference in pixel values ​​at the same point in the two images.

[0101] In the present embodiment, the interior of the shell in the image acquisition device constitutes a darkroom space. When no self-luminous object is placed in it, the photoelectric conversion element first obtains a first dark field image. After the self-luminous object is placed in it, the photoelectric conversion element obtains a second dark field image. The second dark field image is then corrected according to the first dark field image to obtain a target image corresponding to the self-luminous object, thereby obtaining a target image of the self-luminous object with higher clarity through contact imaging. The operation process is simple and the imaging time is short. The imaging device has the advantages of small structure, low manufacturing cost, convenient operation and easy carrying.

[0102] Example 2

[0103] like Figure 7 As shown, the imaging device of the self-luminous object on the biological sample film of this embodiment is a further improvement of the embodiment 1, specifically:

[0104] The imaging device further comprises a light source device 6 , which is arranged in the housing 1 .

[0105] The light source device 6 is arranged at the top position of the inner side of the light shielding cover 4;

[0106] After the light source device 6 is turned on, the corresponding illumination duration is 10ms-30s, and the illumination duration can also be adjusted according to actual needs.

[0107] The light source device 6 includes a plurality of LED lamp beads arranged in a dot matrix, a plurality of lamps introduced by optical fibers, a plurality of lamp tubes arranged in parallel, a plurality of plate-shaped lamps, and the like.

[0108] The imaging device further comprises a light diffusion plate 7 which is fixedly arranged on the inner side of the light shielding cover and is located directly below the light source device, so that the light of the light source device 6 is uniform.

[0109] like Figure 8 As shown, the imaging device further comprises a protective film 8, and two sides of the protective film 8 are respectively attached to the biological sample film and the photoelectric conversion element.

[0110] The thickness of the protective film is 0.01mm-0.2mm, and the material of the protective film is tempered glass film or hard plastic film.

[0111] The outer edge of the protective film 8 is located outside the outer edge of the photoelectric conversion element 2 .

[0112] The photoelectric conversion element 2 includes a probe body 9 and a substrate 10 , and an outer edge of the substrate 10 is located outside the outer edge of the probe body 9 .

[0113] Among them, the substrate is a metal plate, a PCBA (a circuit board manufacturing process) board, etc.

[0114] like Fig. 9 As shown, the top of the base 5 has a receiving groove 11 , and the photoelectric conversion element 2 and the protective film 8 are located in the receiving groove 11 .

[0115] The imaging device of this embodiment further includes a packaging member 12 , which acts on the protective film 8 and the base 5 to limit the movement of the photoelectric conversion element 2 relative to the base 5 .

[0116] Specifically, the package 12 includes a filler 13 and a card board 14 .

[0117] The filler 13 is located in a region surrounded by the photoelectric conversion element 2 , the protective film 8 and the surface of the base 5 , and is used to fix the photoelectric conversion element 2 in the base 5 .

[0118] Among them, the fillers are gel, glass powder, plastic embryos and the like.

[0119] When the photoelectric conversion element 2 is in the packaged state, the clamping plate 14 abuts against the edges of the protective film 8 to ensure the stability of the entire device.

[0120] The base 5 includes a bottom plate 15 and a base body 16. The base body 16 is provided with a receiving groove 11, and the receiving groove 11 penetrates the base body 16. The bottom plate 15 is movably connected to the bottom of the base body 16 to open or close the receiving groove 11 from the bottom of the base body 16. The height of the photoelectric conversion element 2 is not greater than the depth of the receiving groove 11.

[0121] The photoelectric conversion element 2 is also used to obtain a bright field image inside the shell within a set acquisition time after the biological sample membrane is not placed in the shell 1 and the light source device 6 in the darkroom space is turned on, that is, the data acquisition time corresponding to the first dark field image, the second dark field image and the bright field image are all consistent.

[0122] like Fig.10 As shown, the figure is Figure 5 The difference between the first dark field image and the second dark field image is that its overall brightness is higher than the overall brightness of the first dark field image, wherein the fewer the number of oblique lines, the higher the corresponding brightness.

[0123] The image correction device 3 is also used to perform correction processing on the second dark field image according to the first dark field image and the bright field image to obtain a target image corresponding to the self-luminous object.

[0124] The process of acquiring a bright field image using a photoelectric conversion element includes:

[0125] Open the light shielding cover 4 to remove the stains and the like remaining on the stage (i.e., the photoelectric conversion element) in the imaging device;

[0126] Close the light shielding cover 4 so that the interior of the housing 1 forms a darkroom space to ensure that the photoelectric conversion element is not affected by external light signals;

[0127] The light source device is controlled to turn on (the corresponding illumination duration is 10ms-30s), and the photoelectric conversion element 2 is used to synchronously collect the value I of each pixel in the uniform bright field. f1 , where the collection time is 1s.

[0128] The image correction device performs correction processing on the second dark field image according to the first dark field image and the bright field image, and obtains the formula corresponding to the target image corresponding to the self-luminous object as follows:

[0129]

[0130] Where I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d represents the pixel data corresponding to the first dark field image, I f1 Represents pixel data corresponding to a bright field image acquired in a light field provided by a white light lamp in an imaging device.

[0131] The second dark field image, the first dark field image and the bright field image are of the same size, I0-I d The difference and I f1 -I d Both are for the same pixel in the two images, and the ratio I is the percentage of signal intensity corresponding to each pixel.

[0132] Specifically, Fig.11 As shown, the horizontal axis represents the lane (1-5), and the vertical axis represents the signal intensity percentage (unit: %) of each lane, which decreases from left to right. Each lane on the biological sample membrane is selected, and the signal value of each lane is calculated. Based on these signal values, the signal intensity of the target protein in each lane is compared. The larger the signal value, the stronger the signal intensity of the target protein in the corresponding lane, and the darker the displayed color (that is, the clearer the display), as shown in FIG. Fig.12 As shown, from left to right (lanes 1-5) are the locations of the self-luminous objects, i.e., the target proteins, and their corresponding signal intensities gradually decrease, and the displayed colors gradually become lighter. In addition, region B represents the region where the markers of the pre-stained molecular weight standards are located.

[0133] Therefore, according to the signal value obtained by data processing, a more accurate target image of the intensity of the target protein (ie, the self-luminous object) on the biological sample membrane can be obtained.

[0134] In the present embodiment, the interior of the shell in the image acquisition device constitutes a darkroom space. When no self-luminous object is placed in it, the photoelectric conversion element first obtains a first dark field image. After the self-luminous object is placed in it, the photoelectric conversion element obtains a second dark field image. The second dark field image is then corrected according to the first dark field image to obtain a target image corresponding to the self-luminous object. In addition, the bright field image under the light field generated by the light source device in the shell can be combined to more effectively and accurately obtain a target image of the self-luminous object with higher clarity. The operation process is simple and the imaging time is short. The imaging device has the advantages of small structure, low manufacturing cost, convenient operation and easy carrying.

[0135] Example 3

[0136] The imaging device of the self-luminous object on the biological sample film of this embodiment is a further improvement of the embodiment 1, specifically:

[0137] This embodiment includes the light source device 6 in the second embodiment.

[0138] The photoelectric conversion element 2 is also used to obtain a bright field image corresponding to a set acquisition time when the biological sample membrane is not placed in the housing and a uniform light field is provided by an external light source, that is, the data acquisition time corresponding to the first dark field image, the second dark field image and the bright field image are all consistent;

[0139] The image correction device 3 is also used to perform correction processing on the second dark field image according to the first dark field image and the bright field image to obtain a target image corresponding to the self-luminous object, such as Fig.10 As shown, the difference between this image and the first dark field image is that its overall brightness is higher than that of the first dark field image.

[0140] The process of acquiring a bright field image using a photoelectric conversion element includes:

[0141] Open the light shielding cover 4 to remove the stains and the like remaining on the stage (i.e., the photoelectric conversion element) in the imaging device;

[0142] like Fig.13 As shown, in an open space, an external light source (top lighting S) is set just above the imaging device and about 2 meters away from the imaging device;

[0143] Wait for 1 minute, then use the photoelectric conversion element to collect the value I of each pixel in the uniform bright field. f2 .

[0144] The image correction device performs correction processing on the second dark field image according to the first dark field image and the bright field image, and obtains the formula corresponding to the target image corresponding to the self-luminous object as follows:

[0145]

[0146] Where I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d represents the pixel data corresponding to the first dark field image, I f2 Represents pixel data corresponding to a bright field image acquired in a uniform light field provided by an external light source.

[0147] The second dark field image, the first dark field image and the bright field image are of the same size, I0-I d The difference and I f2 -I d Both are for the same pixel in the two images, and the ratio I is the percentage of signal intensity corresponding to each pixel.

[0148] Specifically, Fig.11 As shown, the horizontal axis represents the lane (1-5), and the vertical axis represents the signal intensity percentage (unit: %) of each lane, which decreases from left to right. Each lane on the biological sample membrane is selected, and the signal value of each lane is calculated. Based on these signal values, the signal intensity of the target protein in each lane is compared. The larger the signal value, the stronger the signal intensity of the target protein in the corresponding lane, and the darker the displayed color (that is, the clearer the display), as shown in FIG. Fig.12 As shown, from left to right (lanes 1-5) are the locations of the self-luminous objects, i.e., the target proteins, and their corresponding signal intensities gradually decrease, and the displayed colors also gradually become lighter. In addition, area B represents the area where the markers of the pre-stained molecular weight standards are located.

[0149] Therefore, according to the signal value obtained by data processing, a more accurate target image of the intensity of the target protein (ie, the self-luminous object) on the biological sample membrane can be obtained.

[0150] In the present embodiment, the interior of the shell in the image acquisition device constitutes a darkroom space. When no self-luminous object is placed in it, the photoelectric conversion element first obtains a first dark field image. After the self-luminous object is placed in it, the photoelectric conversion element obtains a second dark field image. The second dark field image is then corrected according to the first dark field image to obtain a target image corresponding to the self-luminous object. In addition, the bright field image under the light field generated by an external light source can be combined to more effectively and accurately obtain a target image of the self-luminous object with higher clarity. The operation process is simple and the imaging time is short. The imaging device has the advantages of small structure, low manufacturing cost, convenient operation and easy carrying.

[0151] Example 4

[0152] like Fig.14 As shown, the imaging method of the self-luminous object on the biological sample film of this embodiment is implemented by the imaging device of the self-luminous object on the biological sample film of embodiment 1, and the imaging method includes:

[0153] S101, using a photoelectric conversion element to obtain a first dark field image of the interior of the inner shell for a set acquisition time;

[0154] The collection time can be set and adjusted according to actual conditions. Preferably, the collection time is set to 1 second.

[0155] S102, after the biological sample film is attached to the surface of the photoelectric conversion element, the photoelectric conversion element is used to obtain a second dark field image inside the housing within a set acquisition time, that is, the data acquisition time corresponding to the first dark field image and the second dark field image is consistent;

[0156] Among them, the biological sample film carries a self-luminous object;

[0157] S103: Correct the second dark field image according to the first dark field image to obtain a target image corresponding to the self-luminous object.

[0158] The formula corresponding to step S103 is as follows:

[0159] I=I0-I d

[0160] Where I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d Indicates pixel data corresponding to the first dark field image.

[0161] The size of the second dark field image is consistent with that of the first dark field image, I0-I d The difference represents the difference in pixel values ​​at the same point in the two images.

[0162] In the present embodiment, the interior of the shell in the image acquisition device constitutes a darkroom space. When no self-luminous object is placed in it, the photoelectric conversion element first obtains a first dark field image. After the self-luminous object is placed in it, the photoelectric conversion element obtains a second dark field image. The second dark field image is then corrected according to the first dark field image to obtain a target image corresponding to the self-luminous object, thereby obtaining a target image of the self-luminous object with higher clarity. The operation process is simple and the imaging time is short. The imaging device has the advantages of small structure, low manufacturing cost, convenient operation and easy carrying.

[0163] Example 5

[0164] like Fig.15 As shown, the imaging method of the self-luminous object on the biological sample film of this embodiment is realized by the imaging device of the self-luminous object on the biological sample film of embodiment 2.

[0165] The imaging method of the self-luminous object on the biological sample film of this embodiment is a further improvement of the embodiment 4, specifically:

[0166] After step S102 and before step S103, the following steps are also included:

[0167] S10301, using a photoelectric conversion element to obtain a bright field image inside the housing within a set acquisition time after the biological sample membrane is not placed in the housing and the light source device in the darkroom space is turned on, that is, the data acquisition time corresponding to the first dark field image, the second dark field image and the bright field image are all the same;

[0168] Step S103 includes:

[0169] S1031, correct the second dark field image according to the first dark field image and the bright field image to obtain a target image corresponding to the self-luminous object. The formula corresponding to step S1031 is as follows:

[0170]

[0171] Where I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d represents the pixel data corresponding to the first dark field image, I f Represents the pixel data corresponding to the bright field image.

[0172] The second dark field image, the first dark field image and the bright field image are of the same size, I0-I d The difference and I f1 -I d Both are for the same pixel in the two images, and the ratio I is the percentage of signal intensity corresponding to each pixel.

[0173] Specifically, Fig.11 As shown, the horizontal axis represents the lane (1-5), and the vertical axis represents the signal intensity percentage (unit: %) of each lane, which decreases from left to right. Each lane on the biological sample membrane is selected, and the signal value of each lane is calculated. Based on these signal values, the signal intensity of the target protein in each lane is compared. The larger the signal value, the stronger the signal intensity of the target protein in the corresponding lane, and the darker the displayed color (that is, the clearer the display), as shown in FIG. Fig.12 As shown, from left to right (lanes 1-5) are the locations of the self-luminous objects, i.e., the target proteins, and their corresponding signal intensities gradually decrease, and the displayed colors gradually become lighter. In addition, region B represents the region where the markers of the pre-stained molecular weight standards are located.

[0174] Therefore, according to the signal value obtained by data processing, a more accurate target image of the intensity of the target protein (ie, the self-luminous object) on the biological sample membrane can be obtained.

[0175] In the present embodiment, the interior of the shell in the image acquisition device constitutes a darkroom space. When no self-luminous object is placed in it, the photoelectric conversion element first obtains a first dark field image. After the self-luminous object is placed in it, the photoelectric conversion element obtains a second dark field image. The second dark field image is then corrected according to the first dark field image to obtain a target image corresponding to the self-luminous object. In addition, the bright field image under the light field generated by the light source device in the shell can be combined to more effectively and accurately obtain a target image of the self-luminous object with higher clarity. The operation process is simple and the imaging time is short. The imaging device has the advantages of small structure, low manufacturing cost, convenient operation and easy carrying.

[0176] Example 6

[0177] like Fig.16 As shown, the imaging method of the self-luminous object on the biological sample film of this embodiment is realized by the imaging device of the self-luminous object on the biological sample film of embodiment 3, and the imaging method of the self-luminous object on the biological sample film of this embodiment is a further improvement of embodiment 4, specifically:

[0178] After step S102 and before step S103, the following steps are also included:

[0179] S10302, using a photoelectric conversion element to obtain a bright field image corresponding to a set acquisition time when the biological sample membrane is not placed in the housing and a uniform light field is provided by an external light source, that is, the data acquisition time corresponding to the first dark field image, the second dark field image, and the bright field image are all consistent;

[0180] Step S103 includes:

[0181] S1032: perform correction processing on the second dark field image according to the first dark field image and the bright field image to obtain a target image corresponding to the self-luminous object.

[0182] The formula corresponding to step S1032 is as follows:

[0183]

[0184] Where I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d represents the pixel data corresponding to the first dark field image, I f Represents the pixel data corresponding to the bright field image.

[0185] The second dark field image, the first dark field image and the bright field image are of the same size, I0-I d The difference and I f2 -I d Both are for the same pixel in the two images, and the ratio I is the percentage of signal intensity corresponding to each pixel.

[0186] Specifically, Fig.11 As shown, the horizontal axis represents the lane (1-5), and the vertical axis represents the signal intensity percentage (unit: %) of each lane, which decreases from left to right. Each lane on the biological sample membrane is selected, and the signal value of each lane is calculated. Based on these signal values, the signal intensity of the target protein in each lane is compared. The larger the signal value, the stronger the signal intensity of the target protein in the corresponding lane, and the darker the displayed color (that is, the clearer the display), as shown in FIG. Fig.12 As shown, from left to right (lanes 1-5) are the locations of the self-luminous objects, i.e., the target proteins, and their corresponding signal intensities gradually decrease, and the displayed colors gradually become lighter. In addition, region B represents the region where the markers of the pre-stained molecular weight standards are located.

[0187] Therefore, according to the signal value obtained by data processing, a more accurate target image of the intensity of the target protein (ie, the self-luminous object) on the biological sample membrane can be obtained.

[0188] In the present embodiment, the interior of the shell in the image acquisition device constitutes a darkroom space. When no self-luminous object is placed in it, the photoelectric conversion element first obtains a first dark field image. After the self-luminous object is placed in it, the photoelectric conversion element obtains a second dark field image. The second dark field image is then corrected according to the first dark field image to obtain a target image corresponding to the self-luminous object. In addition, the bright field image under the light field generated by an external light source can be combined to more effectively and accurately obtain a target image of the self-luminous object with higher clarity. The operation process is simple and the imaging time is short. The imaging device has the advantages of small structure, low manufacturing cost, convenient operation and easy carrying.

[0189] Although the specific embodiments of the present invention are described above, it should be understood by those skilled in the art that these are only examples, and the protection scope of the present invention is defined by the appended claims. Those skilled in the art may make various changes or modifications to these embodiments without departing from the principles and essence of the present invention, but these changes and modifications all fall within the protection scope of the present invention.

Claims

1. An imaging device for a self-luminous object on a biological sample film, characterized in that: The imaging device comprises a housing, a photoelectric conversion element and an image correction device; The interior of the shell forms a darkroom space; The housing comprises a light shielding cover and a base; One side of the light shielding cover is hinged to one side of the base; When the light shielding cover and the base are closed, the interior of the shell forms the darkroom space; The photoelectric conversion element is disposed in the housing; The photoelectric conversion element is used to obtain a first dark field image of the interior of the housing within a set acquisition time when no biological sample membrane is placed therein; Wherein, the biological sample film carries a self-luminous object; After the biological sample film is attached to the surface of the photoelectric conversion element, the photoelectric conversion element is further used to obtain a second dark field image of the interior of the shell within the set acquisition time; The image correction device is used to perform correction processing on the second dark field image according to the first dark field image to obtain a target image corresponding to the self-luminous object; The imaging device further comprises a light source device, and the light source device is arranged in the housing; The photoelectric conversion element is also used to obtain a bright field image inside the housing within the set acquisition time after the biological sample film is not placed in the housing and the light source device in the darkroom space is turned on; Or, the photoelectric conversion element is further used to obtain a bright field image corresponding to the set acquisition time when the biological sample film is not placed in the housing, the housing is in an open state, and a uniform light field is provided by an external light source; The image correction device is also used to perform correction processing on the second dark field image according to the first dark field image and the bright field image to obtain a target image corresponding to the self-luminous object.

2. The imaging device for self-luminous objects on a biological sample film as claimed in claim 1, characterized in that: The image correction device performs correction processing on the second dark field image according to the first dark field image, and obtains the formula corresponding to the target image corresponding to the self-luminous object as follows: I=I0-I d Wherein, I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d Represents pixel data corresponding to the first dark field image.

3. The imaging device of a self-luminous object on a biological sample film as claimed in claim 1, characterized in that: The image correction device performs correction processing on the second dark field image according to the first dark field image and the bright field image, and obtains the formula corresponding to the target image corresponding to the self-luminous object as follows: Wherein, I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d represents the pixel data corresponding to the first dark field image, I f Represents pixel data corresponding to the bright field image.

4. The imaging device of a self-luminous object on a biological sample film as claimed in claim 1, characterized in that: The light source device is arranged at the top position of the inner side of the light shielding cover; and / or, The corresponding illumination duration after the light source device is turned on is 10ms-30s; and / or, The light source device includes a plurality of LED lamp beads arranged in a dot matrix, a plurality of lamps introduced by optical fibers, a plurality of lamp tubes arranged in parallel or a plurality of plate-shaped lamps.

5. The imaging device of a self-luminous object on a biological sample film as claimed in claim 1, characterized in that: The imaging device further comprises a diffuser plate; The light diffuser is fixedly mounted on the inner side of the light shielding cover and is located directly below the light source device; and / or, The photoelectric conversion element includes a CMOS chip, a CCD chip or an amorphous silicon photoelectric conversion detector; and / or, The biological sample membrane includes a protein membrane, an agarose gel block, an agarose gel strip, a polyacrylamide gel block or a polyacrylamide gel strip.

6. The imaging device for self-luminous objects on a biological sample film according to claim 1, characterized in that: The imaging device further includes a protective film; Both sides of the protective film are respectively attached to the biological sample film and the photoelectric conversion element.

7. The imaging device of a self-luminous object on a biological sample film as claimed in claim 6, characterized in that: The thickness of the protective film is 0.01 mm to 0.2 mm; and / or, The material of the protective film is a tempered glass film or a hard plastic film.

8. A method for imaging a self-luminous object on a biological sample film, characterized in that: The imaging method is implemented by using the imaging device of the self-luminous object on the biological sample film according to claim 1, and the imaging method comprises: Using the photoelectric conversion element to acquire a first dark field image of the interior of the housing within a set acquisition time; After the biological sample film is attached to the surface of the photoelectric conversion element, the photoelectric conversion element is used to obtain a second dark field image of the interior of the shell within the set acquisition time; Wherein, the biological sample film carries a self-luminous object; Performing correction processing on the second dark field image according to the first dark field image to obtain a target image corresponding to the self-luminous object; The imaging device also includes a light source device; Before the step of correcting the second dark field image according to the first dark field image to obtain the target image corresponding to the self-luminous object, the step further includes: The photoelectric conversion element is used to obtain a bright field image inside the housing within the set acquisition time after the biological sample film is not placed in the housing and the light source device in the darkroom space is turned on; Or, the photoelectric conversion element is used to obtain a bright field image corresponding to the set acquisition time when the biological sample film is not placed in the housing, the housing is in an open state, and a uniform light field is provided by an external light source; The step of correcting the second dark field image according to the first dark field image to obtain the target image corresponding to the self-luminous object comprises: The second dark-field image is corrected according to the first dark-field image and the bright-field image to obtain a target image corresponding to the self-luminous object.

9. The method for imaging a self-luminous object on a biological sample film according to claim 8, characterized in that: The formula corresponding to the step of correcting the second dark field image according to the first dark field image to obtain the target image corresponding to the self-luminous object is as follows: I=I0-I d Wherein, I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d Represents pixel data corresponding to the first dark field image.

10. The method for imaging a self-luminous object on a biological sample film according to claim 8, characterized in that: The formula corresponding to the step of correcting the second dark-field image according to the first dark-field image and the bright-field image to obtain the target image corresponding to the self-luminous object is as follows: Wherein, I represents the pixel data corresponding to the target image, I0 represents the pixel data corresponding to the second dark field image, and I d represents the pixel data corresponding to the first dark field image, I f Represents pixel data corresponding to the bright field image.

Images