An automated system for high-throughput single cell-based breeding

Abstract

When establishing the most appropriate cells from the huge numbers of a cell library for practical use of cells in regenerative medicine and production of various biopharmaceuticals, cell heterogeneity often found in an isogenic cell population limits the refinement of clonal cell culture. Here, we demonstrated high-throughput screening of the most suitable cells in a cell library by an automated undisruptive single-cell analysis and isolation system, followed by expansion of isolated single cells. This system enabled establishment of the most suitable cells, such as embryonic stem cells with the highest expression of the pluripotency marker Rex1 and hybridomas with the highest antibody secretion, which could not be achieved by conventional high-throughput cell screening systems (e.g., a fluorescence-activated cell sorter). This single cell-based breeding system may be a powerful tool to analyze stochastic fluctuations and delineate their molecular mechanisms.

Classification of C2C12 cells at differentiation by convolutional neural network of deep learning using phase contrast images

“…The vertical column on the left represents the image size, and the horizontal column above represents the size of the kernel. For example, [3][4][5][6] indicates that the kernel size of the first convolution layer is 3 × 3 and the kernel size of the second convolution layer is 6 × 6. As a combination of kernel sizes, a total of nine ways are implemented.…”

“…To realize regenerative medicine, industrialization of tissue/organ regenerations is one important topic because the industrialization is expected to reduce human errors and labor cost, which provides safety, security and affordable tissue/organ supply. Tissue/organ regeneration requires large numbers of cells, typically 1 cm 3 tissue include about 10 9 cells, so that automatic cell-culturing system has been developed [3,4] and already commercially available. The next target must be developing the method how to evaluate cellular conditions in culturing state for further tissue/organ engineering.…”

System Integration, Modelling, and Simulation for Automation of Multiple Microfluidic Stream Based Bio-manipulation

“…In other previous of automatic bio-manipulation method [16]- [20] in micro fluidic environment, as the target of Bio-manipulation is cells which are extremely small in size, susceptible, and sensitive, precisely automatic control lead to great advantage. However, most of these automatic systems functioned for closed space, fixed pattern and single function micro fluidic system.…”

Automated single cell sorting and deposition in submicroliter drops

“…2(d)). Anomalous pressure of silicon oil was in the range given by the manufacturer for the vapor pressure at [20][21][22][23][24][25] C. Evaporation turned to be a major factor that has to be considered in microliter scale drop deposition processes. To overcome the problem of the critical dropping pressure (sum of the Laplace pressure, vapor recoil and pressure due to vapor extension) we opened both Valve 1 and Valve 2 (Fig.…”

“…24 A similar robot for the automated breeding of single cells has been recently developed. 25 This integrated instrument applies microwell arrays to immobilize cells on the surface for subsequent sorting. We consider our system being introduced in the current letter for single cell isolation and deposition more accessible for research and medical diagnostics as it can be mounted onto any standard inverted microscope available in most laboratories.…”