diff --git a/content/13.hardware.md b/content/13.hardware.md
index 9e48fe768851fa9aa470e555a15f94a2123b09c5..dc3ff291581c264a958fc06249deabc859f8c147 100644
--- a/content/13.hardware.md
+++ b/content/13.hardware.md
@@ -10,58 +10,67 @@
## **Introduction**
-Having determined that we wanted to design a gate switch for oxygen and lactic acid, we were thinking about how to detect the expression intensity in different oxygen environments and different lactic acid concentrations. A relatively mature measurement method is to use ELISA, but ELISA cannot control the concentration of lactic acid and oxygen. In the actual experiment, we cultured it in a stable environment for a period of time and then transferred it to ELISA for measurement. The fluorescence intensity data measured by the enzyme labeler also need to exclude the influence of the concentration of bacterial solution. In the actual experiment, the fluorescence intensity was divided by OD value. With the progress of the experiment, although we used the macroscopic measurement means of ELISA to get differentiated results, a more accurate measurement can make our data more reliable. We eventually plan to use microfluidic chip technology to obtain single-cell precision fluorescence intensity data at precise oxygen and lactate concentrations.
+After deciding to design an AND-gate for sensing hypoxia and high lactic acid, we consider how to detect the expression intensity in different oxygen environments and various lactic acid concentrations. A relatively established measurement method is ELISA, but Multi-Mode Microplate Reader (Biotek) lacks control over the concentration of lactic acid and oxygen. In our actual experiment, we culture E. coli in a stable environment for a specific duration before transferring it to Biotek for measurement. The fluorescence intensity data obtained from Biotek also require the exclusion of bacterial culture concentration's influence. In the experiment, the fluorescence intensity is divided by the OD value. We obtain differentiated results by Biotek, but Biotek can only perform fluorescence intensity measurements at a macroscopic level, with relatively lower accuracy. Additionally, the plasmids we introduced into E. coli are of low copy number, resulting in inherently low fluorescence intensity. This increases the measurement error in our results. A more accurate measurement method can enhance the reliability of our data. Our ultimate plan is to employ microfluidic chip technology to acquire single-cell precise fluorescence intensity data at specific oxygen and lactate concentrations.
## **Design of the microfluidic chip**
-Microfluidic chips need to meet the following functions:
+Microfluidic chip need to meet the following functions:
1, There is an oxygen concentration gradient in the chip environment
2, The chip can bind the bacteria to grow in a small area
-3, the chip simultaneously measures a variety of bacteria, and does not interfere with each other
+3, the chip simultaneously measures a variety of E.coli, and the E.coli do not interfere with each other
-Based on the above three requirements, we made changes on the basis of the previous chip and designed a chip that can form six oxygen concentrations and measure four kinds of bacteria at the same time [Figure 1].。
+Based on the above three requirements, we make changes on the basis of the previous chip and design a chip that forms six oxygen concentrations and measure four types of E.coli simultaneously (see Figure 1).
<br/>
<center><img src="w1.png" alt="Figure 1" style="width: 70%;" /></center>
-
<font size=3 color=grey>
-
-**Figure 1** The above figure shows the details of the culture layer (Mold 1) and loading layer (Mold 2). The culture layer (Mold 1) is composed of 3 masks on the left, and the loading layer (Mold 2) on the right is composed of 2 mask. To clearly show the detailed pattern of each layer of the two molds, the size in the figure does not represent the true proportion of the structure. All our masks are drawn using L-edit software, and the tdb format file is also uploaded in [https://gitlab.igem.org/2023/software-tools/peking](https://gitlab.igem.org/2023/software-tools/peking)
+**Figure 1** The above figure shows the details of the culture layer (Mold 1) and loading layer (Mold 2). The culture layer (Mold 1) is composed of 3 masks on the left, and the loading layer (Mold 2) on the right is composed of 2 mask. To clearly show the detailed pattern of each layer of the two molds, the size in the figure does not represent the true proportion of the structure. All our masks are drawn using L-edit software, and the tdb format file is uploaded in [https://gitlab.igem.org/2023/software-tools/peking](https://gitlab.igem.org/2023/software-tools/peking)
</font>
<br/>
-We designed the trap to be very low, only 1.3 microns, and the chamber outlet is also very narrow, only 3 microns, so once the E. coli enters the chamber, it is trapped in it. The method of feeding bacteria and culture medium is shown in Figure 2.
+We design the chamber to be very low, only 1.3 microns, and the chamber outlet is also very narrow, only 3 microns, so when the E. coli enters the chamber, it becomes trapped in it. The method of loading bacterial culture and culture solution is shown in Figure 2.
<br/>
<center><img src="w2.png" alt="Figure 1" style="width: 70%;" /></center>
-
+**Figure 2** The method of loading. Step 1 load the E.coli, Step 2 load the solution and flush the E. coli into the trap.
<br/>
+The first step of loading is to plug the culture inlet. The bacteria solution flows in from the bacteria inlet and out through the bacteria outlet. At this stage, a large number of E.coli will be distributed in the flow channel. The second step of loading is to plug the bacteria inlet and open the culture inlet. The culture solution will pass through the trap due to the pressure difference, thereby flushing the E. coli into the trap. This method not only allows real-time updating of the culture fluid in the trap but also enables trapping of the E.coli.
+
+
-We first blocked the mouth into the culture medium, the bacterial liquid flowed in from the inlet and out of the outlet. At this time, there will be a large number of bacteria distributed in the flow channel. Then, the outlet is blocked and the outlet of the culture liquid is opened, and the culture liquid will pass through the trap due to the pressure difference, thus flushing the E. coli in the flow path into the trap. In this way, not only can the culture fluid in the trap be updated in real time, but also the E. coli can be bound in the trap.
+## **Manufacture process of the microfluidic chip**
+We first use laser printing to create a mask consisting of five layers of chips (see Figure 3a). Next, we apply a certain height of photoresist and expose it to develop a mold consisting of two layers of chips (see Figure 3b and Figure 3c). PDMS is then spread on the mold to obtain the upper and lower chips, which are subsequently spliced together (see Figure 3d). The chip is then baked overnight in an oven at 70℃. The following day, the chip is punched (see Figure 3e) and combined with the slide (see Figure 3f). Finally, the chip is baked for over 3 hours.
<br/>
+<br/>
+
+<center>
+
+<img src="w100.png" alt="Figure 1" style="width: 70%;" />
-<center><img src="w3.jpg" alt="Figure 1" style="width: 70%;" /></center>
+</center>
+<font size=3 color=grey>
+**Figure 3** Manufacture process. (a) 5 masks. (b1) and (c1) loading layer mold and its microgragh. (b2) and (c2) culture layer mold and its microgragh. (d) combination of two layer. (e) Baking the combined layer overnight, followed by the need for punching. (f) combine layer with the slide.
<br/>
@@ -81,7 +90,6 @@ We tested the feasibility of the chip and proved that our chip can bind E. coli
**Figure 4** The growth of E. coli in the trap chamber
-
</font>
<br/>
@@ -102,21 +110,18 @@ Due to the defects of the air pump, the concentration gradient is not stable, bu
<center><img src="w9.png" alt="Figure 1" style="width: 70%;" /></center>
-
<font size=3 color=grey>
**Figure 5** Fluorescence intensity of 6 channels
-
</font>
<br/>
-
Using the Stern-Volmer equation,
-$I_0/I=1+K_q [O_2]$
+$$I_0/I=1+K_q [O_2]$$
where $[O_2]$ represents the concentration of oxygen, $I$ represents the fluorescence intensity at this concentration, $I_0$ represents the fluorescence intensity in the absence of oxygen, and $K_q$ is the Stern-Volmer quenching constant. We can get the fluorescence intensity $I_0$ and $I_{21}$ at 0% and 21% by passing only nitrogen and air, and then calculate $K_q$, and get the corresponding relationship between oxygen concentration and fluorescence intensity:
- $[O_2 ]=(I_0/I -1)/(I_0/I_{21} -1)×21\%$
+ $$[O_2 ]=(I_0/I -1)/(I_0/I_{21} -1)×21\%$$
Due to lack of time, we did not do a quantitative oxygen concentration calculation.
@@ -128,10 +133,14 @@ $I_0/I=1+K_q [O_2]$
<font size=3 color=grey>
-**Figure 6** The change of fluorescence intensity after standardization
-
+**Figure 6** The change of fluorescence intensity after standardization.
</font>
<br/>
+## Conclusion
+
+We design and successfully construct a microfluidic chip, and detect the chip's ability to trap Escherichia coli and to facilitate the creation of oxygen concentration gradients. Due to time constraints, we have not used the chip for measuring bacterial fluorescence intensity, but this chip is proven to be feasible. Our chip also allows for experimental measurements of expression variation caused by different lactate concentrations in the culture medium.
+
+Our chip has a wide range of application scenarios where fluorescence measurements targeting E.coli can be conducted on it, while enabling precise control over oxygen concentration and culture medium concentration.
diff --git a/content/14.design.md b/content/14.design.md
index 7d92d091dc01fcb112e52016ee41ed8a55607c94..d98ed3166687b0c7d361caacd2733e2582328455 100644
--- a/content/14.design.md
+++ b/content/14.design.md
@@ -145,7 +145,7 @@ In addition, in our design, the expression of vesicles is not controlled by the
<br>
-## **Controllable enveloping**
+## **Controllable encapsulation**
<br>
diff --git a/content/15.results.md b/content/15.results.md
index 556bf6104da53a07746f3a8927853e535f59a52d..ed2019d2372aaefec9f32bfa0304e433aadfbaa1 100644
--- a/content/15.results.md
+++ b/content/15.results.md
@@ -96,7 +96,7 @@ In order to measure fluorescence intensity with single cell accuracy at precise
<br>
-<center><img src="r2/chipschematic.jpg" alt="Figure 2" style="width: 70%;"/></center>
+<center><img src="r2/chipschematic2.jpg" alt="Figure 2" style="width: 70%;"/></center>
<font size=3 color=grey>