1. Bill Trogler
UCSD

2. Current Biosensing Cannot Readily Detect Individual Cancer Cells in the Body1
1000
1 million
1 billion
1 trillion or 40 generations
Number of cancer cells

3. Hollow Nanoparticles A Biomimetic Synthetic Analogue of a Virus for Cell Delivery and Sensing
200 nm SiO nm SiO nm TiO2

4. Silica like virus particles readily enter living cells with Davorka Messmer’s group
Uptake of FITC-labeled 100 nmSiO2 NPs by primary human dendritic cells (40x)
45nm-FITC nanoparticles are taken up by dendritic cells and traffic to the nucleus (Confocal microscopy 60x)

5. Targeted Delivery to Endothelial Cells with Carson and Esener groups
100 nm amine modified silica hollow spheres with CGKRK targeting peptide uptake by HUVEC cells vs. nontargeting CREKA control

6. Hollow Nanosphere Cell Biosensors with A. Kummel and S. Esener’s groups
Interior contains pH sensitive
Fluorophore, redox sensitive
Fluorophore, temperature
Sensing quantum dot, ….
TEM image

7. D. Messmer, A. C. Kummel, W. C. Trogler UCSD
The Single Cell Hyper Analyzer for Secretants (SiCHAS)
D. Messmer, A. C. Kummel, W. C. Trogler UCSD
Arrays of microwells will be coated with a mixture of secretant detection antibodies. Secretant molecules bound to the antibodies on the bottom of the wells will be detected by fluorescently labeled secondary antibodies with different emission spectra.
Cell surface molecules will be simultaneously detected with quantum dot labeled antibodies.
The wells will be connected with narrow slits to allow the cells to communicate via secretants or bigger slits to allow physical cell-cell contact.
Note the wells will actually be at least 10x taller than the cells (100 micron) but only 20x20 microns in-plane to limit out diffusion; they are shown as short wells here just for graphical clarity.

8. D. Messmer, A. C. Kummel, W. C. Trogler UCSD
The Single Cell Hyper Analyzer for Secretants (SiCHAS)
D. Messmer, A. C. Kummel, W. C. Trogler UCSD
(TOP) Cell seeding in microwells. 50 x 50 x 50 μm wells 10μm thickwalls. 50,000 cells were spread over 1cm2 so we had less than one cells/well initially. The cells divide over time so there density increases. Green is a membrane stain.
(Bottom Left) Proof of Viability: The cells were kept in the wells sealed by a coverslip for 5 days in culture media. After 5 days, a small amount of propidium iodide (PI) was added which stains dead cells red. The image on the left shows there are almost no dead cells.
(Bottom Right) Proof of Diffusion: If a mixture of dead and live cells is used and stain with PI, many of the cells are red proving we can diffuse stain to the bottom of the wells. The wells employed for this experiment were 60x60x50 μm nl/well.

9. Specific Absorption of Breast Cancer Cells MCF-7 – with Dr
Specific Absorption of Breast Cancer Cells MCF-7 – with Dr. Sarah Blair, A. Kummel’s groups
Gold Coverage=8.73%
Cells on Substrate=21
Cells on Gold Pads=16=76.19%
Coverage Ratio: = 8.73 x random
Selective binding of breast cancer cell line tested. (Above) Bright yellow circles are cells and dim red circles are the gold pads. (Right) Cell nucleus is blue. Cell membrane is green. Au pad is red.
Gold pads represent only 9% of the surface area but 76% of the cells attached to the gold pads. This is 9x random.
Increased selectivity will be obtained by improvement of the washing step.

10. Grand Challenges Viral-like endocytosed intracellular biosensors
Sensing cell-cell signaling in ordered cell arrays to capture biodiversity of population
Parallel sensing equivalent of flow cytometry on ordered arrays of cells
Cell specific preconcentration