Abstract
Oocyte enucleation is a critical procedure for somatic cell nuclear transfer. Yet, the main steps of oocyte enucleation are still manually operated, which presents several drawbacks such as low precision, high repetition error, and long training time for operators. For improving the operation efficiency and success rate, a robotic precise oocyte blind enucleation method is presented in this paper. The proposed method involves the following key techniques: oocyte translation control, oocyte immobilization and penetration control, and enucleation volume control based on the adaptive slide mode. Compared with the manual blind enucleation method, the proposed robotic blind enucleation method reduced the operation time by 44.5% (manual method: 62 s vs. proposed method: 34.4 s), increased the accuracy of enucleation by 83.1% (manual method: 30.7 vs. proposed method: 5.2), increased the success rate from 80% to 93.3%, and increased the cleavage rate from 41.7% to 63.3%.
Highlights
Cloning refers to the process in which cells develop into new organisms through asexual reproduction [1,2]
The proposed robotic enucleation method estimated the overshoot compared with the other control methods
A pair of XYZ micromanipulators were used for mounting the holding micropipette and the enucleation micropipette (EM), and a host computer was used for microscopic image processing and motion control of the XY stage, manipulators, and syringe pump
Summary
Cloning refers to the process in which cells develop into new organisms through asexual reproduction [1,2]. In 1997, British scientist Wilmut et al used sheep mammary cells as donor cells for somatic cell nuclear transfer (SCNT) and obtained the world’s first somatic cloned sheep, Dolly. The standard nuclear transplantation procedure is divided into two parts: the enucleation of oocytes and the injection of donor cells. The enucleation operation needs to tear apart the genetic material and cytoplasm in oocytes, which is the most difficult step in the operation of somatic cell nuclear transfer (SCNT). The main steps of oocyte enucleation are still manually operated [9], which presents several drawbacks, for example, low precision, high repetition error, and long training time for operators [10]. It is necessary to automate the operation process of oocyte enucleation [11,12,13], which will improve the operation precision of oocyte enucleation, improve the success rate of oocyte enucleation, and further promote micro-manipulation technology and gene technology [14]
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