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Automated Nanoscale Absolute Accuracy Alignment System for Transfer Printing.

ABSTRACT: The heterogeneous integration of micro- and nanoscale devices with on-chip circuits and waveguide platforms is a key enabling technology, with wide-ranging applications in areas including telecommunications, quantum information processing, and sensing. Pick and place integration with absolute positional accuracy at the nanoscale has been previously demonstrated for single proof-of-principle devices. However, to enable scaling of this technology for realization of multielement systems or high throughput manufacturing, the integration process must be compatible with automation while retaining nanoscale accuracy. In this work, an automated transfer printing process is realized by using a simple optical microscope, computer vision, and high accuracy translational stage system. Automatic alignment using a cross-correlation image processing method demonstrates absolute positional accuracy of transfer with an average offset of <40 nm (3? < 390 nm) for serial device integration of both thin film silicon membranes and single nanowire devices. Parallel transfer of devices across a 2 × 2 mm2 area is demonstrated with an average offset of <30 nm (3? < 705 nm). Rotational accuracy better than 45 mrad is achieved for all device variants. Devices can be selected and placed with high accuracy on a target substrate, both from lithographically defined positions on their native substrate or from a randomly distributed population. These demonstrations pave the way for future scalable manufacturing of heterogeneously integrated chip systems.

SUBMITTER: McPhillimy J

PROVIDER: S-EPMC7590505 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Automated Nanoscale Absolute Accuracy Alignment System for Transfer Printing.

McPhillimy John J Jevtics Dimitars D Guilhabert Benoit J E BJE Klitis Charalambos C Hurtado Antonio A Sorel Marc M Dawson Martin D MD Strain Michael J MJ

ACS applied nano materials 20200923 10

The heterogeneous integration of micro- and nanoscale devices with on-chip circuits and waveguide platforms is a key enabling technology, with wide-ranging applications in areas including telecommunications, quantum information processing, and sensing. Pick and place integration with absolute positional accuracy at the nanoscale has been previously demonstrated for single proof-of-principle devices. However, to enable scaling of this technology for realization of multielement systems or high thr ...[more]

PMID: 33134883

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Project description:BackgroundGene transfer is an important tool for cellular therapies. Lentiviral vectors are most effectively transferred into lymphocytes or hematopoietic progenitor cells using spinoculation. To enable cGMP (current Good Manufacturing Practice)-compliant cell therapy production, we developed and compared a closed-system spinoculation method that uses cell culture bags, and an automated closed system spinoculation method to decrease technician hands on time and reduce the likelihood for microbial contamination.MethodsSepax spinoculation, bag spinoculation, and static bag transduction without spinoculation were compared for lentiviral gene transfer in lymphocytes collected by apheresis. The lymphocytes were transduced once and cultured for 9 days. The lentiviral vectors tested encoded a CD19/CD22 Bispecific Chimeric Antigen Receptor (CAR), a FGFR4-CAR, or a CD22-CAR. Sepax spinoculation times were evaluated by testing against bag spinoculation and static transduction to optimize the Sepax spin time. The Sepax spinoculation was then used to test the transduction of different CAR vectors. The performance of the process using healthy donor and a patient sample was evaluated. Functional assessment was performed of the CD19/22 and CD22 CAR T-cells using killing assays against the NALM6 tumor cell line and cytokine secretion analysis. Finally, gene expression of the transduced T-cells was examined to determine if there were any major changes that may have occurred as a result of the spinoculation process.ResultsThe process of spinoculation lead to significant enhancement in gene transfer. Sepax spinoculation using a 1-h spin time showed comparable transduction efficiency to the bag spinoculation, and much greater than the static bag transduction method (83.4%, 72.8%, 35.7% n = 3). The performance of three different methods were consistent for all lentiviral vectors tested and no significant difference was observed when using starting cells from healthy donor versus a patient sample. Sepax spinoculation does not affect the function of the CAR T-cells against tumor cells, as these cells appeared to kill target cells equally well. Spinoculation also does not appear to affect gene expression patterns that are necessary for imparting function on the cell.ConclusionsClosed system-bag spinoculation resulted in more efficient lymphocyte gene transfer than standard bag transductions without spinoculation. This method is effective for both retroviral and lentiviral vector gene transfer in lymphocytes and may be a feasible approach for gene transfer into other cell types including hematopoietic and myeloid progenitors. Sepax spinoculation further improved upon the process by offering an automated, closed system approach that significantly decreased hands-on time while also decreasing the risk of culture bag tears and microbial contamination.

Dataset Information

Automated Nanoscale Absolute Accuracy Alignment System for Transfer Printing.

Publications

Automated Nanoscale Absolute Accuracy Alignment System for Transfer Printing.

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