Supporting Information for:
Supporting Information for: Capillary Flow Layer-by-Layer: A Microfluidic Platform for the High Throughput Assembly and Screening of Nanolayered Film Libraries Steven A. Castleberry1,2,3,†, Wei Li1,2,5,†, Di Deng4, Sarah Mayner1 & Paula T. Hammond1,2,* 1
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, Koch Institute of Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 3 Harvard-MIT Division of Health Sciences and Technology, Cambridge, MA 4 Department of Materials Science and Engineering, Massachusetts Institute of Technology, Cambridge, MA 5 Current address: Department of Chemical Engineering, Texas Tech University, Lubbock, TX † These authors contributed equally to this work. 2
Corresponding author: Paula T. Hammond Professor, Department of Chemical Engineering Koch Institute of Integrative Cancer Research Massachusetts Institute of Technology Room 76-553 Cambridge, MA 02139 Email: [email protected]
The PDF file includes: Supplementary Figure S1. Treatment Supplementary Figure S2. Supplementary Figure S3. Supplementary Figure S4. Supplementary Figure S5. Supplementary Figure S6. Transfection Efficacy
Photolithographic Mask Showing Partial O2 Plasma CF-LbL Device Arrangement is Modular and Customizable Automated CF-LbL Assembly Manual CF-LbL Assembly The Effect of PAH pH on Cell Density Cell Spread Area and Cell Number Compared to
Supplementary Figure S1. Photolithographic Mask Showing Partial O2 Plasma Treatment. The shaded portion of the design is covered during O2 plasma treatment so that it remains hydrophobic. This maintains the separation of material between channels to reduce the chance of cross-contamination during assembly.
Supplementary Figure S2: CF-LbL Device Arrangement is Modular and Customizable. (a) Each channel is independent of neighboring channels. Channel spacing is designed to pair with multichannel pipettes. (b) The device can be operated where channels are connected with an in-device manifold system (G 1.5 & G 2.0) or are held completely separate (G 1.0). Shaded regions denote the portion of the PDMS mold that is covered during plasma treatment so as to remain hydrophobic. (c) The CF-LbL channels can be arranged to 384-well plate spacing with 96 independent channels contained in the space of a common well-plate. This design consists of three modular units (A-B, C-D, and E-F) which can each be placed on individual microscope slides.
Supplementary Figure S3. Automated CF-LbL Assembly. Repeated material adsorption and washing steps for the automated operation of the CF-LbL device. A constant vacuum is applied to the device, however it only acts on the main channel when both the capillary flow break well (2) and the exit well (3) are covered.
Supplementary Figure S4. Manual CF-LbL Assembly Method. This schematic describes the bench top application of the CF-LbL device. The operation of this simple device requires only that the PDMS be sealed to a suitable substrate (e.g. glass, silicon, plastic, etc.) and that the user have a means of placing a droplet of working solution into the entrance well and a source of compressed air to assist in channel clearance. Capillary force ensures that the entire channel is filled with all wetting solutions and that excess material solutions can be recovered.
Supplementary Figure S5. The Effect of PAH pH on Cell Density. (a) The impact of PAH assembly pH on initial seeding of cells on a surface. (b) Increasing PAH assembly pH corresponds with decreased cell number after seven days. A clear change is apparent between pH 6.5 and 7.5. This is likely due to a reduction in the degree of ionization of the PAH at these conditions. Error bars represent 95% confidence interval.
Supplementary Figure S6. Cell Spread Area and Cell Number Compared to Transfection Efficacy. (a) Fraction GFP positive cells graphed with the corresponding average cell spread area. No trend can be seen. (b) Fraction GFP positive cells versus the number of cells on a given surface. No trend in transfection was seen for either criteria.
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, Koch Institute of Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 3 Harvard-MIT Division of Health Sciences and Technology, Cambridge, MA 4 Department of Materials Science and Engineering, Massachusetts Institute of Technology, Cambridge, MA 5 Current address: Department of Chemical Engineering, Texas Tech University, Lubbock, TX † These authors contributed equally to this work. 2
Corresponding author: Paula T. Hammond Professor, Department of Chemical Engineering Koch Institute of Integrative Cancer Research Massachusetts Institute of Technology Room 76-553 Cambridge, MA 02139 Email: [email protected]
The PDF file includes: Supplementary Figure S1. Treatment Supplementary Figure S2. Supplementary Figure S3. Supplementary Figure S4. Supplementary Figure S5. Supplementary Figure S6. Transfection Efficacy
Photolithographic Mask Showing Partial O2 Plasma CF-LbL Device Arrangement is Modular and Customizable Automated CF-LbL Assembly Manual CF-LbL Assembly The Effect of PAH pH on Cell Density Cell Spread Area and Cell Number Compared to
Supplementary Figure S1. Photolithographic Mask Showing Partial O2 Plasma Treatment. The shaded portion of the design is covered during O2 plasma treatment so that it remains hydrophobic. This maintains the separation of material between channels to reduce the chance of cross-contamination during assembly.
Supplementary Figure S2: CF-LbL Device Arrangement is Modular and Customizable. (a) Each channel is independent of neighboring channels. Channel spacing is designed to pair with multichannel pipettes. (b) The device can be operated where channels are connected with an in-device manifold system (G 1.5 & G 2.0) or are held completely separate (G 1.0). Shaded regions denote the portion of the PDMS mold that is covered during plasma treatment so as to remain hydrophobic. (c) The CF-LbL channels can be arranged to 384-well plate spacing with 96 independent channels contained in the space of a common well-plate. This design consists of three modular units (A-B, C-D, and E-F) which can each be placed on individual microscope slides.
Supplementary Figure S3. Automated CF-LbL Assembly. Repeated material adsorption and washing steps for the automated operation of the CF-LbL device. A constant vacuum is applied to the device, however it only acts on the main channel when both the capillary flow break well (2) and the exit well (3) are covered.
Supplementary Figure S4. Manual CF-LbL Assembly Method. This schematic describes the bench top application of the CF-LbL device. The operation of this simple device requires only that the PDMS be sealed to a suitable substrate (e.g. glass, silicon, plastic, etc.) and that the user have a means of placing a droplet of working solution into the entrance well and a source of compressed air to assist in channel clearance. Capillary force ensures that the entire channel is filled with all wetting solutions and that excess material solutions can be recovered.
Supplementary Figure S5. The Effect of PAH pH on Cell Density. (a) The impact of PAH assembly pH on initial seeding of cells on a surface. (b) Increasing PAH assembly pH corresponds with decreased cell number after seven days. A clear change is apparent between pH 6.5 and 7.5. This is likely due to a reduction in the degree of ionization of the PAH at these conditions. Error bars represent 95% confidence interval.
Supplementary Figure S6. Cell Spread Area and Cell Number Compared to Transfection Efficacy. (a) Fraction GFP positive cells graphed with the corresponding average cell spread area. No trend can be seen. (b) Fraction GFP positive cells versus the number of cells on a given surface. No trend in transfection was seen for either criteria.
