SUPPORTING INFORMATION A Framework to Account for ...
SUPPORTING INFORMATION A Framework to Account for Sedimentation and Diffusion in Particle-Cell Interactions Jiwei Cui,†,# Matthew Faria,†,‡,# Mattias Björnmalm,† Yi Ju,† Tomoya Suma,† Sylvia T. Gunawan,† Joseph J. Richardson,†,§ Hamed Heidari,∆ Sara Bals,∆ Edmund J. Crampin,‡ and Frank Caruso†,* †
ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, and the
Department of Chemical and Biomolecular Engineering, the University of Melbourne, Parkville, Victoria 3010, Australia; ‡ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, and the Systems Biology Laboratory, Melbourne School of Engineering, The University of Melbourne, Parkville, Victoria 3010, Australia; ∆Electron Microscopy for Materials Research (EMAT), University of Antwerp, Groenenborgerlaan 171, 2020 Antwerp, Belgium. #
These authors contributed equally.
§
Current address: CSIRO Manufacturing Flagship, Clayton, Victoria 3168, Australia.
1
Fluorescence Intensity (a.u.)
1.2x107 1.0x10
7
Y=1.91627E8X+852958.9 R2=0.9995
8.0x106 6.0x106 4.0x106 2.0x106 0.00
0.01
0.02
0.03
0.04
0.05
-1
Concentration (mg mL )
Figure S1. Calibration curve of AF488-labeled PMAPDA.
2
Figure S2. Design of support for inverted particle-cell-association experiments. Support is placed at the bottom of a well in a 12-well culture plate. A glass slide with cultured cells is placed upside-down (i.e., cells facing down) on top of pillars. 3D model file (.skp file, for SketchUp software) is available from the author.
3
Figure S3. Design of support for vertical particle-cell-association experiments. A glass slide (with cultured cells) is slid into the slits so that the cells are facing sideward. The slits are 1 mm by 1 mm in cross-section and go for the full length of the chamber plus an additional 1 mm into the bottom of the chamber. 3D model file (.skp file, for SketchUp software) is available from the author.
4
Figure S4. TEM images of (a) SC/MS particles before pore enlargement and (b) APTESmodified SC/MS particles with enlarged pores.
5
Figure S5. Scheme of cross-linking strategy in MS templates to prepare SC/MS@PMA particles and PMA capsules.
6
Figure S6.
Different viewing directions of volume renderings of the tomographic
reconstruction of PMA capsules obtained from a tilt series of cryo-TEM images.
7
Figure S7. Ultramicrotomed TEM image of PMA capsules. The apparent heterogeneity in particle size is due to slicing of particles through random sections.
8
Cell viability (%)
100 80 60 40 20 0 Upright
Inverted
Vertical
Figure S8. Cell viability after 24 h of the cells positioned in the upright, inverted, and vertical orientations.
9
Figure S9. Super-resolution microscopy images with maximum intensity projection of HeLa cells after 24 h incubation with (a,b) SC/MS@PMA particles or (c,d) PMA capsules in the (a,c) upright or (b,d) inverted orientations. Blue, green and red represent nuclei stained with Hoechst, actin stained with AF488, and particles or capsules labeled with AF647, respectively.
10
Figure S10. 2D concentration profiles within wells for SC/MS@PMA particles. From the top, concentration profiles are: Without cells, with cells on the bottom of the plate wells (upright), with cells on the top (inverted), with cells on the side (vertical). Upright and inverted orientations were modeled using a 1D partial differential equation (PDE), but a 2D plot is shown for visual clarity.
11
Figure S11. 2D concentration profiles within cell-plate wells for PMA capsules. From the top, concentration profiles are: Without cells, with cells on the bottom of the plate wells (upright), with cells on the top (inverted), with cells on the side (vertical). Upright and inverted orientations were modeled using a 1D PDE, but a 2D plot is shown for visual clarity.
12
Table S1. Size, shell thickness, zeta potential, and average density of SC/MS@PMA particles and PMA capsules. Sample SC/MS@PMA particles PMA capsules
Diameter (nm) 250 ± 10
Shell thickness (nm) 38 ± 3
Zeta potential (mV) -38.6 ± 6
Average density (mg mL-1) 1.65
200 ± 10
30 ± 2
-35.2 ± 5
1.06
13
Table S2. Raw values for theoretical cellular dose. Samples SC/MS@PMA particles PMA capsules
Upright 0.374 0.106
Inverted 0.015 0.084
Vertical 0.048 0.054
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ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, and the
Department of Chemical and Biomolecular Engineering, the University of Melbourne, Parkville, Victoria 3010, Australia; ‡ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, and the Systems Biology Laboratory, Melbourne School of Engineering, The University of Melbourne, Parkville, Victoria 3010, Australia; ∆Electron Microscopy for Materials Research (EMAT), University of Antwerp, Groenenborgerlaan 171, 2020 Antwerp, Belgium. #
These authors contributed equally.
§
Current address: CSIRO Manufacturing Flagship, Clayton, Victoria 3168, Australia.
1
Fluorescence Intensity (a.u.)
1.2x107 1.0x10
7
Y=1.91627E8X+852958.9 R2=0.9995
8.0x106 6.0x106 4.0x106 2.0x106 0.00
0.01
0.02
0.03
0.04
0.05
-1
Concentration (mg mL )
Figure S1. Calibration curve of AF488-labeled PMAPDA.
2
Figure S2. Design of support for inverted particle-cell-association experiments. Support is placed at the bottom of a well in a 12-well culture plate. A glass slide with cultured cells is placed upside-down (i.e., cells facing down) on top of pillars. 3D model file (.skp file, for SketchUp software) is available from the author.
3
Figure S3. Design of support for vertical particle-cell-association experiments. A glass slide (with cultured cells) is slid into the slits so that the cells are facing sideward. The slits are 1 mm by 1 mm in cross-section and go for the full length of the chamber plus an additional 1 mm into the bottom of the chamber. 3D model file (.skp file, for SketchUp software) is available from the author.
4
Figure S4. TEM images of (a) SC/MS particles before pore enlargement and (b) APTESmodified SC/MS particles with enlarged pores.
5
Figure S5. Scheme of cross-linking strategy in MS templates to prepare SC/MS@PMA particles and PMA capsules.
6
Figure S6.
Different viewing directions of volume renderings of the tomographic
reconstruction of PMA capsules obtained from a tilt series of cryo-TEM images.
7
Figure S7. Ultramicrotomed TEM image of PMA capsules. The apparent heterogeneity in particle size is due to slicing of particles through random sections.
8
Cell viability (%)
100 80 60 40 20 0 Upright
Inverted
Vertical
Figure S8. Cell viability after 24 h of the cells positioned in the upright, inverted, and vertical orientations.
9
Figure S9. Super-resolution microscopy images with maximum intensity projection of HeLa cells after 24 h incubation with (a,b) SC/MS@PMA particles or (c,d) PMA capsules in the (a,c) upright or (b,d) inverted orientations. Blue, green and red represent nuclei stained with Hoechst, actin stained with AF488, and particles or capsules labeled with AF647, respectively.
10
Figure S10. 2D concentration profiles within wells for SC/MS@PMA particles. From the top, concentration profiles are: Without cells, with cells on the bottom of the plate wells (upright), with cells on the top (inverted), with cells on the side (vertical). Upright and inverted orientations were modeled using a 1D partial differential equation (PDE), but a 2D plot is shown for visual clarity.
11
Figure S11. 2D concentration profiles within cell-plate wells for PMA capsules. From the top, concentration profiles are: Without cells, with cells on the bottom of the plate wells (upright), with cells on the top (inverted), with cells on the side (vertical). Upright and inverted orientations were modeled using a 1D PDE, but a 2D plot is shown for visual clarity.
12
Table S1. Size, shell thickness, zeta potential, and average density of SC/MS@PMA particles and PMA capsules. Sample SC/MS@PMA particles PMA capsules
Diameter (nm) 250 ± 10
Shell thickness (nm) 38 ± 3
Zeta potential (mV) -38.6 ± 6
Average density (mg mL-1) 1.65
200 ± 10
30 ± 2
-35.2 ± 5
1.06
13
Table S2. Raw values for theoretical cellular dose. Samples SC/MS@PMA particles PMA capsules
Upright 0.374 0.106
Inverted 0.015 0.084
Vertical 0.048 0.054
14
