## 3 Results
### Analysis of Blood Cell lysis Efficiency  
We confirmed the efficiency of blood cell lysis via image analysis and counting the number of cells before and after cell lysis. For comparison, we also analyzed the number of cells in the control group without an applied electric field. Representative analyzed images are shown in Figure 4a,b. As shown in Figure 4a, images of the control group without the cell lysis were obtained at 20s intervals from the microchannel near the Nafion membrane. For images of the blood cell lysis caused by ICP, before and after images of the  
Figure 3: Continuous lysis of blood cells in the developed microfluidic device. (**a,b**) Continuous injection of diluted blood without applied electric field: (**a**) Brightfield image showing blood cells flowing near the Nafion membrane. (**b**) Timelapse sequence image magnifying the dashed region in (**a**). Eight consecutive frames are shown, captured every 33 ms (total duration: 264 ms). Blood cells maintain their shape and remain intact throughout the sequence, demonstrating that no lysis occurs in the absence of an electric field. (**c,d**) Continuous injection of diluted blood with an electric field applied (75 V): (**c**) Brightfield image showing blood cells near the Nafion membrane under electric field. (**d**) Timelapse sequence magnifying the dashed region in (**c**). Eight consecutive frames are shown at 33 ms intervals. The blood cell progressively loses its defined shape, becomes increasingly transparent, and by the final frame (264 ms), is indistinguishable from the background—indicating successful lysis. Black arrows indicate the temporal sequence of frames. (all images were acquired at a flow rate of 1000 nL/min and 100 ms exposure time) (scale bar in Figure 3b,d: white bar in 33 ms image, 100 μm).  
microchannel near the Nafion membrane were obtained without and with the electric field applied for 1 min. As shown in Figure 4b, concentrated blood cell lysates were observed in all cases. The number of blood cells was measured using ImageJ software (version 1.53e) with an exposure time of 100 ms at the time of capture. As shown in Figure 4c, the number of blood cells dramatically decreased after lysis, with approximately 99.4% \(\pm\) 0.2% (n = 5) of the cells injected into the microchannel lysed.  
To evaluate the operational boundaries of our device, we experimentally examined how variations in flow rate and applied voltage affect cell lysis performance. As shown in Figure S1a, increasing the flow rate from 1 to 3 \(\mu\)L/min while maintaining the applied voltage between 75 and 100 V resulted in a substantial decrease in lysis efficiency. This is attributed to the higher drag force on the cells, which causes them to transit through the depletion region too quickly to experience effective electric field disruption. Additionally, as shown in Figure S1b, lowering the applied voltage from 75 V to 40 V at a constant flow rate of 1 \(\mu\)L/min also led to negligible lysis, indicating that a minimum threshold field strength is necessary to maintain disruption capability. These results highlight a trade-off between throughput (flow rate) and operating voltage. Based on these findings, effective lysis is consistently achieved at flow rates up to 1 \(\mu\)L/min with applied voltages of 75 V or higher. Operating beyond these ranges--such as at flow rates above 3 \(\mu\)L/min or voltages below 40 V--markedly reduces lysis efficiency and may introduce challenges such as biofouling or bubble formation (Figure S2). This analysis offers guidance for optimizing device operation for different application scenarios and sample types.