Application Cases

Experiment Name: Experimental Study on Ultrasonic Detachment Technology for Adherent Cells

Research Direction:
Design and adaptation of ultrasonic vibration systems, feasibility validation of cell detachment technology, and optimization of key parameters for detachment technology.

Experimental Objective:
This study investigates an ultrasonic vibration-based method for adherent cell detachment. An ultrasonic transducer unit was designed considering both the automation requirements of detachment applications and experimental needs. Experiments were conducted using control parameters such as input signal, liquid medium in the flask, and distance/position as variables to explore their effects on detachment efficiency. The optimal control parameters for achieving the best detachment performance were determined.

Testing Equipment:

1. Ultrasonic Vibration System: Control center, signal generator, high-voltage power amplifier (ATA-4052), oscilloscope (DZ4000), ultrasonic transducer unit, linear slide, stepper motor, holder. The experimental block diagram is shown in Figure 1.

2. Cell Experiment Equipment: Bright-field microscope, cell incubator, centrifuge, T75 culture flasks, trypan blue detection equipment.

3. Auxiliary Equipment and Consumables: Various culture media, physiological saline, serum substitutes, penicillin-streptomycin, various growth factors, etc.

   
   

Figure 1: Ultrasonic Vibration System

Experimental Procedure:
The ultrasonic vibration system consists of a control center, signal generation, signal detection, motion control, and ultrasonic vibration modules. The control center manages the signal generation and motion control components. The signal generator produces the required experimental signal, which is then amplified by a high-power amplifier (ATA-4052, Aigtek, China) and monitored by an oscilloscope (DZ4000, Rigol, China) for signal verification before being transmitted to the ultrasonic transducer unit.

The ultrasonic transducer unit comprises four ultrasonic transducers (40k60, Guangyuanda, China) arranged in a square configuration. A linear slide moves the cell culture flask, adjusting the distance d between the adherent cell layer and the transducer to position the cells within the ultrasonic energy field. A groove placed on the ultrasonic transducer holds the ultrasonic coupling medium.

Experimental Results:

1. Determination of Optimal Parameters:
   When the ultrasonic transducer input voltage was set to 255 V, with a frequency sweep of 159–161 kHz, an ultrasonic exposure time of 2 minutes, a cell-to-transducer distance of 3 mm, and 12 mL of physiological saline as the vibration liquid, the cell detachment area in the T75 flask reached 95%, cell viability was 70%, and the detachment efficiency parameter K reached 67%.

2. Influence of Various Parameters:

• (1) Voltage: Cell detachment rate was positively correlated with voltage, while cell viability was negatively correlated. Increasing voltage improved detachment efficiency but also increased cell damage (see Figure 2).

• (2) Frequency: No linear relationship was observed between cell detachment rate and frequency; however, higher frequencies resulted in higher cell viability. The optimal detachment effect was achieved around 160 kHz. Within the same frequency range, frequency-swept pulses yielded higher detachment rates than continuous or burst pulses (see Figure 3).

• (3) Distance: The optimal detachment effect was observed at a cell-to-transducer distance of 3 mm. A shorter distance (2 mm) increased cell damage due to physical contact, while a longer distance (4 mm) reduced the detachment rate (see Figure 4).

• (4) Liquid Medium in Flask: Pure physiological saline provided the best detachment performance, while media-containing liquids resulted in lower detachment rates. When the liquid volume was 12 mL (corresponding to a height of 1.6 mm), the optimal balance between detachment rate and cell viability was achieved (see Figure 5).

  
  

Figure 2: Effect of Voltage on Cell Detachment

Figure 3: Effect of Frequency on Cell Detachment

Figure 4: Effect of Distance on Cell Detachment

Figure 5: Effect of Liquid Medium in Flask on Cell Detachment

Product Recommendation: ATA-4052C High-Voltage Power Amplifier

Figure: ATA-4052C High-Voltage Power Amplifier Specifications and Parameters

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