TWI531785B - Detective device and detective method - Google Patents

Description

Detection device and detection method

The invention relates to a detecting device and a detecting method, and relates to a detecting device and a detecting method for detecting a state of a biochemical sample.

Cell culture is an indispensable technology in basic biology and medical research. Generally, the cells are cultured in a culture dish containing the culture medium, and the cell culture medium is periodically replaced and the cells are subcultured. Regular and close observation of cell status is an important step in cell culture as cells will continue to grow, proliferate, and produce metabolites, and may be subject to infections such as viruses or bacteria during culture.

Existing cell culture operations rely primarily on manual processing. However, since the growth state and proliferation rate of the cells may vary with time and environment, and there may be unexpected events such as cell infection and culture fluid deterioration, it is difficult for the operator to change the culture fluid. Conduct immediate judgment and disposal. Furthermore, the calculation of cell density is a cumbersome step for the operator. Therefore, if an automated instrument can help determine the state of the cell, it will help to improve the quality and efficiency of cell culture.

The invention provides a state detecting device for a biochemical sample to obtain an instant state of a biochemical sample.

The invention further provides a detection method to obtain an immediate state of a biochemical sample.

The detecting device of the present invention can be used to detect the state of the biochemical sample, and the detecting device comprises a loading platform, a first light source, a second light source, a light sensing component and a control component. The stage is used to carry a biochemical sample. The first light source emits a first wavelength of light to illuminate the biochemical sample at a first time point. The second source emits a second wavelength of light and illuminates the biochemical sample at a second time point. The light sensing component is configured to receive the first wavelength light and the second wavelength light after passing through the biochemical sample to obtain a first read value and a second read value, respectively. The control component has a first reference value corresponding to the light of the first wavelength and a second reference value corresponding to the light of the second wavelength, and is used for comparing the first read value with the first reference value And a second read value and a second reference value.

The detecting method of the present invention is for detecting the state of a biochemical sample, and the detecting method comprises the following steps. Obtaining a first reference value and a second reference value. A biochemical sample is irradiated with light of a first wavelength, and the first wavelength of light after passing through the biochemical sample is received by the photo sensing element to obtain a first read value. The biochemical sample is irradiated with the second wavelength light, and the second wavelength light after passing through the biochemical sample is received by the photo sensing element to obtain a second read value. Comparing the first read value with the first reference value to perform a first processing action. Comparing the second read value with the second reference value to perform a second processing action.

Based on the above, the detecting device and the detecting method of the present invention have different color lights Biochemical samples are irradiated in fractions to facilitate detection of different characteristics of biochemical samples. In this way, by synthesizing the characteristics of the biochemical samples obtained, the state of the biochemical samples can be accurately and immediately determined.

In order to make the above-described features of the present invention more comprehensible, the following detailed description of the embodiments will be described in detail below.

10‧‧‧Detection and Processing System

100‧‧‧Detection device

110‧‧‧stage

120‧‧‧First light source

130‧‧‧second light source

140‧‧‧Light sensing components

150‧‧‧Control elements

160A‧‧‧First Processing Unit

160B‧‧‧Second processing unit

170‧‧‧ Display elements

180‧‧‧Power supply

A, B‧‧‧ area

BS‧‧‧ biochemical sample

S210~S248‧‧‧Steps

FIG. 1A is a schematic diagram of a detecting device according to an embodiment of the invention.

FIG. 1B is a schematic diagram of a light sensing element of a detecting device according to an embodiment of the invention.

2 is a flow chart of a detection method in accordance with an embodiment of the invention.

FIG. 3 is a flow chart of a detection method according to an embodiment of the invention.

4 is a flow chart of a detection method in accordance with an embodiment of the invention.

FIG. 5 is a flow chart of a detection method according to an embodiment of the invention.

FIG. 6 is a flow chart of a detection method according to an embodiment of the invention.

7 is a schematic diagram of a detection and processing system in accordance with an embodiment of the present invention.

8A is a photographic view of the cell culture solution of Samples 1 to 8 of the experimental example, and FIG. 8B is an OD value measured by a spectrophotometer when the cell culture solution is irradiated with the first wavelength light and the second wavelength light, respectively. Line chart.

9A-9D are judgments of a first state in accordance with an embodiment of the present invention. Schematic diagram of the breaking method.

10A and FIG. 10B are schematic diagrams showing the principle of a second state determination method according to an embodiment of the invention.

FIG. 1A is a schematic diagram of a detecting device according to an embodiment of the invention. The detecting device 100 is configured to detect a state of the biochemical sample, and includes a stage 110, a first light source 120, a second light source 130, a light sensing component 140, and a control component 150. The stage 110 is used to carry the biochemical sample BS. The stage 110 is made of, for example, a light transmissive material such as plastic or glass. The biochemical sample BS contains a culture solution and a culture body, for example, a cell or a protein, which is placed in a carrier and cultured in a culture solution or suspended in a solution. The culture solution is, for example, a light-transmitting liquid having a color or a color. In addition, the culture solution usually changes color due to a change in pH, so deterioration of the culture solution itself or excessive cell metabolites or infection may cause discoloration of the culture solution. For example, taking the culture solution as DMEM, the culture solution changes from pink to orange depending on the pH value. Furthermore, as the cell content or protein content increases, the overall transmittance of the biochemical sample BS decreases.

The first light source 120 emits a first wavelength of light to illuminate the biochemical sample BS at a first time point. The second light source 130 emits a second wavelength of light to illuminate the biochemical sample BS at a second time point. That is, the first light source 120 and the second light source 130 respectively illuminate the biochemical sample BS at different times. The second wavelength light is different from the wavelength of the first wavelength light. In this embodiment, the wavelength range of the first wavelength light is, for example, 600 nm to 780. Nm, which is for example red light. The wavelength of the second wavelength light is, for example, 400 nm to 600 nm, which is, for example, green light or yellow light. In the present embodiment, the biochemical sample BS is placed between the first light source 120 and the second light source 130 and the light sensing element 140, for example. In detail, the first light source 120 and the second light source 130 are disposed, for example, above the biochemical sample BS, and the photo sensing element 140 is disposed under the biochemical sample BS. As a result, the light emitted by the first light source 120 or the second light source 130 is detected by the light sensing element 140 after passing through the biochemical sample BS. The first light source 120 and the second light source 130 are, for example, irradiated in the same area of the biochemical sample BS. In this embodiment, the first light source 120 and the second light source 130 are substantially the same light source, and can be switched to emit the first wavelength light or the second wavelength light by adjusting the wavelength, but the invention is not limited thereto. For example, in other embodiments, the first light source 120 and the second light source 130 may also be two members respectively, and then the desired light source is moved over the biochemical sample BS by, for example, rotating, so that the first light source 120 The same area as the biochemical sample BS is irradiated with the second light source 130. In addition, although in the present embodiment, the first light source 120 and the second light source 130 are directly above, the first light source 120 and the second light source 130 may be disposed at any position capable of illuminating the biochemical sample BS. For example, lateral illumination or the like is performed by the first light source 120 and the second light source 130, respectively, and the light sensing element 140 can sense the light intensity passing through the biochemical sample BS. Of course, in other embodiments, the first light source 120 and the second light source 130 may be irradiated to different regions of the biochemical sample BS as needed. It should be noted that the culture solution in the present embodiment is DMEM, and the first light source 120 and the second light source 130 respectively use red light and green light to respectively correspond to pink and orange colors formed by the pH change of the culture liquid. In other embodiments, according to The difference in color produced by the different samples can adjust the wavelength range of the first wavelength light and the wavelength range of the second wavelength light to implement the detection method of the present invention.

The light sensing component 140 is configured to receive the first wavelength light and the second wavelength light after passing through the biochemical sample BS to obtain a first read value and a second read value, respectively. In the present embodiment, the light sensing element 140 has, for example, different responses to light of different wavelengths, and the light sensing element 140 is, for example, a reaction intensity of light of the second wavelength light higher than that of the first wavelength light. In the present embodiment, the light sensing component 140 is, for example, a single sensor (as shown in FIG. 1A) or an array sensor (as shown in FIG. 1B), wherein the array sensor can be used to calculate biochemical samples. The area of the distribution area is, for example, an Amorphous Selenium Active Pixel Sensor (a-Se APS). In another embodiment, the light sensing element 140 can also be a thin film transistor element, a CCD, a CMOS, a photodiode, or the like.

The light sensing element 140 converts the light source passing through the biochemical sample BS into an electrical signal. In the present embodiment, the light sensing element 140 is disposed under the stage 110, for example, but the invention is not limited thereto. For example, in another embodiment, the light sensing elements can be integrated into the stage.

The control component 150 is configured to receive the first read value and the second read value from the light sensing component, and has a built-in light sensing component corresponding to a first reference value of the first wavelength light and corresponding to the first A second reference value of the two-wavelength light. Therefore, the control element 150 compares the first read value with the first reference value, and compares the second read value with the second reference value, and outputs the comparison result. In this embodiment, the control component 150 is further connected to the first light source 120 and the second light source 130, for example. The biochemical sample BS is irradiated with the first light source 120 or the second light source 130 by control.

Next, the detection method of the present invention will be described in conjunction with a detection device. 2 is a flow chart of a detection method in accordance with an embodiment of the invention. Referring to FIG. 1A and FIG. 2 simultaneously, first, a correction step is performed on the light sensing element 140 (step S210). The calibration step is respectively in two stages, the first stage is to establish a first reference value of the first wavelength of the biochemical sample corresponding to the first wavelength, and the second stage is to establish the second wavelength of the biochemical sample corresponding to the second wavelength of light a second reference value, wherein the first reference value and the second reference value are generated according to a density, a concentration, a quantity, an area of the distribution area, a pH value, or a color of the biochemical sample. A first biochemical standard is provided in a first state, such as a DMEM culture medium that exhibits a pink color. Irradiating the first biochemical standard with the first wavelength light (step S212), and receiving the first wavelength light after passing the first biochemical standard by the light sensing element 140 to obtain the first reference value L1 (step S214), The first reference value L1 is then set in the control element 150 (step S216). The wavelength of the first wavelength light is, for example, 600 nm to 780 nm.

A second biochemical standard in a second state is provided, the second biochemical standard comprising, for example, a DMEM broth that turns orange due to deterioration of the culture fluid or infection of the culture. The second biochemical standard is irradiated with the second wavelength light (step S218), and the second wavelength light after passing the second biochemical standard is received by the light sensing element 140 to obtain the second reference value L ₂ (step S220) And the second reference value L _{2 is} set in the control element 150 (step S222). The wavelength of the second wavelength light ranges from 400 nm to 600 nm.

After performing the above-described correcting step on the light sensing element 140, the photometric sensing element 140 is used to perform a detection step for the first wavelength light on the biochemical sample (step S230). In detail, the biochemical sample BS is irradiated with the first wavelength light (step S232), and the first wavelength light after passing through the biochemical sample BS is received by the photo sensing element 140 to obtain the first read value L ₁ ' (step S234). Transmitting the first read value L ₁ ' to the control element 150, the control element 150 comparing the first read value L ₁ ' with the first reference value L ₁ to confirm the condition that the biochemical sample corresponds to the first state (step S236) . In the present embodiment, the first read value L ₁ ' rises as the cell density increases. The comparison method may be to directly compare the first read value L ₁ ' with the first reference value L ₁ or calculate OD ₁ , OD ₁ = L ₁ / L ₁ '. When the first read value L ₁ ' is smaller than the first reference value L ₁ , that is, L ₁ '<L ₁ , the control element 150 sends a signal that can continue to cultivate the biochemical sample BS, and the signal is transmitted to the first light source 120, for example. To monitor the biochemical sample BS by irradiating the biochemical sample BS with the first wavelength light again after a specific time interval. When the first read value L ₁ ' is greater than the first reference value L ₁ , that is, L ₁ '>L ₁ , the control component 150 issues a signal to immediately perform an action such as a splitting on the biochemical sample BS, such as Transfer to the processing unit to perform the first process (step S238).

For example, when the culture body in the biochemical sample BS is a cell, the first reference value L ₁ may be a critical cell density, and when the first read value L ₁ ' exceeds the first reference value L ₁ , it indicates that the cell needs to be performed. Distribute processing. Furthermore, the first reference value L ₁ may be a value directly read by the light sensing element 140 or a calculated ratio, and the calculated ratio may be advantageous for removing background value differences such as machine error. Further, the density of the culture body in the first biochemical standard can be adjusted and step S214 is repeated to generate a plurality of first reference values. For example, a density of 50%, 60%, or 70% of the culture can be used as the first biochemical standard to obtain a corresponding plurality of first reference values, or 80%, 90% of the culture body can be used. Or 100%, etc., the density of the discs must be immediately performed to obtain a corresponding plurality of first reference values.

In step S240, the biosensor sample BS is subjected to a detection step for the second wavelength light using the light sensing element 140 (step S240). In detail, the biochemical sample BS is irradiated with the second wavelength light (step S242), and the photo sensing element 140 receives the second wavelength light after passing through the biochemical sample BS, and obtains the second read value L ₂ ' (step S244). Next, the second read value L ₂ ' is transmitted to the control element 150, and the control element 150 compares the second read value L ₂ ' with the second reference value L ₂ to confirm the condition of the biochemical sample corresponding to the second state ( Step S246). In the present embodiment, the second read value L ₂ ' decreases as the degree of discoloration of the culture solution increases. The comparison method may be to directly compare the second read value L ₂ ' with the second reference value L ₂ or calculate OD ₂ , OD ₂ = L ₂ / L ₂ '. When the second read value L ₂ ' is less than the second reference value L ₂ , that is, L ₂ '<L ₂ , the control element 150 determines to continue to irradiate the biochemical sample BS with the second wavelength light after a certain time interval to monitor biochemistry. Sample BS. When the second read value L ₂ ' is greater than the second reference value L ₂ , that is, L ₂ '>L ₂ , the control element 150 issues a need to replace the culture solution with the biochemical sample BS, add the drug to the culture solution, or discard the cells. The processed signal, for example, is transmitted to the processing unit for performing the second processing (step S248). The second reference value L ₂ may be a value directly read by the light sensing component 140 or a calculated ratio, wherein the calculated ratio may be beneficial to remove background value differences such as machine error.

In an embodiment, as shown in FIG. 10A, since the intensity of the reaction of the light sensing element 140 with respect to the second wavelength light (B region) is higher than the reaction intensity of the first wavelength light (A region), according to the intensity of the received light Change trend can judge the correspondence of biochemical samples In the second state, that is, whether the culture solution is discolored or not. For example, as shown in FIG. 10B, when the measured brightness value rises, the rising speed of the light absorption value (OD) is slowed down, and even the turning point is lowered (as indicated by the dotted circle), which can be reasonably speculated. The reason for this is that the region where the reaction intensity is shifted to the light sensing element 140 is high, indicating that the color of the culture solution changes.

It should be noted that the present invention does not limit the sequence of steps described above. For example, as shown in FIG. 3, the first read value and the second read value of the biochemical sample BS pair may also be measured first ( That is, steps S232, S234 and S242, S244), and then the status of the biochemical sample BS corresponding to the first state and the second state is determined (ie, steps S236 and S246), and finally the first and the first corresponding to the biochemical sample BS are performed. Two processes (ie steps S238 and S248). Alternatively, as shown in FIG. 4, after the first read value and the second read value of the biochemical sample BS are measured (ie, steps S232, S234 and S242, S244), step S236 is performed separately or simultaneously. S246 interpretation and processing steps. Alternatively, in an embodiment, the detecting step of S242 and S244 and the processing step of S246 may be performed before the detecting step of S232 and S234 and the processing step of S246. Furthermore, in an embodiment, as shown in FIG. 5, step S210 may already be built into the detecting device, so step S210 may also be omitted.

In one embodiment, shown in Figure 6, reference may be made to a third predetermined value L ₃ (step S215), the third reference value L ₃ is measured continuously over time a first reference value L ₁ The speed of change obtained. The third reference value L _{3 is} used to compare with the change speed of the plurality of first read values L ₁ ' (ie, the third read value L ₃ ') to confirm the change speed of the biochemical sample BS corresponding to the first state (Step S215). The third reference value L ₃ can be a critical cell growth rate. In detail, the first reference value L ₁ is measured by measuring the first reference value L ₁ twice at least at two time points to obtain a predetermined third reference value L ₃ . . Similarly, a predetermined fourth reference value L ₄ may be obtained (step S223), and the fourth reference value L ₄ is a rate of change obtained by continuously measuring the second reference value L ₂ for a certain period of time. The fourth reference value L _{4 is} used to align with the rate of change of the plurality of second read values L ₂ ' (ie, the fourth read value L ₄ ') to confirm the rate of change of the biochemical sample corresponding to the second state. In detail, the second reference value L ₂ is measured by measuring the second reference value L ₂ twice at least at two time points to obtain a predetermined fourth reference value L ₄ . For example, the fourth reference value L ₄ is, for example, a critical speed indicating a color change of the culture liquid, and therefore exceeding the fourth reference value L ₄ indicates, for example, that the color change speed of the culture liquid is too fast and it is necessary to immediately change the liquid. It should be noted that, as described above, the present invention does not limit the above-described sequence of steps, so the third reference value L ₃ , the fourth reference value L ₄ , the third read value L ₃ ' and the fourth read value The acquisition of L ₄ ' can be combined with the aforementioned sequence of steps of FIGS. 3 to 5.

7 is a schematic diagram of a detection and processing system in accordance with an embodiment of the present invention. Referring to FIG. 7, the detection and processing system 10 incorporates the aforementioned detecting apparatus 100 shown in FIGS. 1A and 1B and is applicable to the aforementioned detecting method. The detection and processing system 10 includes a first light source 120, a second light source 130, a light sensing component 140, a control component 150, a first processing unit 160A, a second processing unit 160B, a display component 170, and a Power supply 180. The control element 150 is connected to the first light source 120, the second light source 130, and the light sensing element 140. For detailed description, reference may be made to the previous embodiment, and details are not described herein. In the present embodiment, the control element 150 is, for example, connected to the display element 170, and the display element 170 is, for example, a display surface. A board or a display with an alarm function to display the comparison result on the panel as a message or to give a warning tone to provide the status of the biochemical sample BS to the operator.

In the present embodiment, the control element 150 is, for example, further connected to the processing unit (such as the first processing unit 160A and the second processing unit 160B. The control element 150 is compared with the first state and the second state according to the biochemical sample BS. As a result, the processing unit is instructed to perform processing such as splitting, changing the culture solution, adding the drug, or discarding the biochemical sample BS. In the present embodiment, the first process is a process in which the number of cells is excessive, and the control element 150 can send a message to notify the disk. Or send a message to inform the removal, or send a message to inform the liquid change, the liquid change includes the water absorption process by the first processing unit 160A and the water injection or the administration by the second processing unit 160B. The second process is the color change process. The control component 150 can also instruct the first processing unit 160A or the second processing unit 160B to automatically perform related processing for issuing a message.

Next, an experimental example will be given to illustrate that the second wavelength light of the present invention can be used to detect changes in the color of the cell culture medium. 8A is a photographic view of the cell culture solution of Samples 1 to 8 of the experimental example, and FIG. 8B is an OD value measured by a spectrophotometer when the cell culture solution is irradiated with the first wavelength light and the second wavelength light, respectively. Line chart. The coloration of the cell culture solution of Samples 1 to 8 was changed from pink to yellow in color, wherein the cell culture medium was DMEM, and cells were not placed in Samples 1 to 8, and only different colors were exhibited by changes in pH. The wavelength of the first wavelength light ranges from 600 nm to 780 nm, and the wavelength of the second wavelength light ranges from 400 nm to 600 nm. Please refer to FIG. 8A and FIG. 8B at the same time, and the experimental example shows that the color of the cell culture liquid has When the sample 1 to the sample 8 are changed, the OD value of the cell culture liquid for the first wavelength light is substantially similar, but the OD value of the second wavelength light is drastically decreased. That is to say, the color change of the cell culture fluid can be extracted by the combination of the first wavelength light and the second wavelength light.

It is to be noted that although the color of the culture liquid is used as the distinction between the first state and the second state in the above embodiment, the present invention is not limited thereto. The first state and the second state include the density, concentration, quantity, and area of the distribution area of the biochemical sample (as shown in FIGS. 9A to 9D, wherein FIG. 9D reaches the disc standard), and the pH of the culture solution for culturing the biochemical sample Or the color of the culture solution. In other words, the light sensing element can be widely used to detect various characteristics related to the optical sample. In addition, in the above embodiment, the cell density detection is performed on the biochemical sample first, and then the culture liquid state detection is taken as an example, but the present invention is not limited to this order. In addition, although the two types of wavelength light are taken as an example in the above embodiment, in other embodiments, two or more types of wavelength light may be used, that is, two or more types of features may be detected.

In summary, the detecting device and the detecting method of the present invention sequentially irradiate the biochemical samples by different color lights to separately detect different characteristics of the biochemical samples. In one embodiment, the light sensing component has a greater reaction intensity for the first wavelength light and the second wavelength light, that is, the light sensing component has better sensing properties for the two types of light, thereby improving light sensing. The ability of a component to detect specific features of a biochemical sample. In this way, by synthesizing the characteristics of the biochemical samples obtained, the state of the biochemical samples can be judged more accurately and immediately, and the biochemical samples can be appropriately processed.

For example, due to the growth state and proliferation rate of cells over time It is different from the changes in the environment, and there may be unexpected events such as cell infection and deterioration of the culture fluid, so it is difficult for the operator to immediately judge and handle the changes in the culture fluid. If the detecting device of the embodiment of the present invention is used, the cell density and the color of the culture solution can be easily monitored, and the state of the cells can be known. In this way, it will be beneficial to greatly improve the quality and efficiency of cell culture. Of course, the present invention can be widely applied to other biochemical related technologies, such as the Western blot method to detect protein concentration, thereby improving research tools in the biochemical field. In other words, the present invention can be applied to detect various optically relevant features in a biochemical sample to facilitate obtaining or monitoring the state of the biochemical sample.

Although the present invention has been disclosed in the above embodiments, it is not intended to limit the present invention, and any one of ordinary skill in the art can make some changes and refinements without departing from the spirit and scope of the present invention. The scope of the invention is defined by the scope of the appended claims.

S210~S248‧‧‧Steps

Claims (15)

A detecting device for detecting a state of a biochemical sample, comprising: a stage for carrying the biochemical sample; a first light source emitting a first wavelength light to illuminate the biochemical sample at a first time point a second light source emitting a second wavelength of light to illuminate the biochemical sample at a second time point; a light sensing element for receiving the first wavelength light and the second wavelength light after passing the biochemical sample , respectively, generating a first read value and a second read value; and a control component having a first reference value of the biochemical sample in a first state and the biochemical sample in a second state a second reference value for comparing the first read value with the first reference value and comparing the second read value with the second reference value. The detecting device of claim 1, wherein the first reference value corresponds to a density, a concentration, a quantity, an area of a distribution area, a pH value or a color of the biochemical sample in the first state, the first The two reference values correspond to the density, concentration, number, area of the distribution area, pH value or color of the biochemical sample in the second state. The detecting device of claim 1, wherein the light sensing element has a higher response intensity to the second wavelength light than the first wavelength light. The detecting device of claim 1, wherein the light sensing The component respectively generates a plurality of the first read value and the plurality of second read values; the control component further includes a third reference value and a fourth reference value for comparing the third reference value with the a rate of change of the read value, and a rate of change of the fourth reference value and the second read values. The detecting device according to claim 1, wherein the biochemical sample comprises a culture solution and a culture body, and the culture body is a cell or a protein. The detecting device of claim 1, wherein the light sensing element is an array element for calculating an area of a distribution area of the biochemical sample. The detecting device of claim 2, wherein the wavelength range of the first wavelength light corresponds to a color of the biochemical sample in the first state, and the wavelength range of the second wavelength light corresponds to the biochemical sample The color of the second state. A detecting method for detecting a state of a biochemical sample, comprising: obtaining a first reference value and a second reference value; illuminating the biochemical sample with a first wavelength light, and receiving by a light sensing component Passing the first wavelength light after the biochemical sample to obtain a first read value; irradiating the biochemical sample with the second wavelength light, and receiving the second after passing the biochemical sample by the light sensing element Wavelength light to obtain a second read value; comparing the first read value with the first reference value to perform a first processing action; and comparing the second read value with the second reference value to Perform a second processing action. The detecting method of claim 8, wherein the method of obtaining the first reference value comprises irradiating a first biochemical standard with the first wavelength light, and Receiving, by the light sensing component, the first wavelength light after passing the first biochemical standard; the method for obtaining the second reference value comprises: irradiating a second biochemical standard with the second wavelength light, and by using the The light sensing element receives the second wavelength light after passing the second biochemical standard. The detecting method of claim 8, wherein the light sensing component receives the first wavelength light after passing the biochemical sample to obtain a plurality of the first read values; the detecting method further comprises Comparing the rate of change of the first read values with a third reference value. The detecting method of claim 8, wherein the light sensing component receives the second wavelength light after passing the biochemical sample to obtain a plurality of the second read values; the detecting method further comprises Comparing the rate of change of the second read values with a fourth reference value. The detecting method of claim 8, wherein the light sensing element has a higher reaction intensity to the second wavelength light than the first wavelength light. The detection method according to claim 8, wherein the biochemical sample comprises a culture solution and a culture, and the culture is a cell, a culture solution or a protein. The detection method of claim 8, wherein the first reference value corresponds to a density, a concentration, a quantity, an area of a distribution area, a pH value or a color of the biochemical sample in a first state; The two reference values correspond to the density, concentration, quantity, area of the distribution area, pH value or color of the biochemical sample in a second state. The detecting method of claim 14, wherein the first wave The wavelength range of the long light corresponds to the color of the biochemical sample in the first state, and the wavelength range of the second wavelength light corresponds to the color of the biochemical sample in the second state.