Methods of non-destructive nanostraw intracellular sampling for longitudinal cell monitoring
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsNicholas A. MELOSH, Yuhong CAO, Karl Martin HJORT, Amanda JONSSON
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Abstract
Methods and apparatuses to non-destructively and periodically sample a small quantity of intracellular proteins and mRNA from the same single cell or cells for an extended period of time. Specifically, describe herein are non-perturbative methods for time-resolved, longitudinal extraction and quantitative measurement of intracellular proteins and nucleic acids from a variety of cell types using systems including nanostraws.
Claims21
(canceled)
A system for nondestructively sampling intracellular material, the system comprising:
- a cell culture chamber having an upper region and a lower region;
- a nanostraw substrate positioned over the lower region, wherein the substrate comprises a plurality of sample regions;
- a plurality of nanostraws extending through the nanostraw substrate in each sample region, wherein each nanostraw has an outer diameter configured to support a cell;
- a plurality of sample material collectors, wherein each sample material collector corresponds to one sample region of the plurality of sample regions;
- a first electrode in the upper region;
- a second electrode in the lower region;
- and
- a controller coupled to the first electrode and the second electrode and configured to apply a pulsed voltage through the plurality of nanostraws of between about 1 V and 100V, a pulse width of between about 10 microseconds and 50 milliseconds for a duration of between 1 second and 300 seconds.
A system for nondestructively sampling intracellular material, the system comprising:
- a cell culture chamber having an upper region and a lower region;
- a nanostraw substrate positioned over the lower region, wherein the substrate comprises a pattern of recessed sample regions;
- a plurality of nanostraws extending through the nanostraw substrate in each sample region, wherein each nanostraw has an outer diameter configured to support a cell without penetrating a cell's cell membrane, wherein the outer diameter is between about 20 nm to about 1500 nm;
- a plurality of removable sample material collectors comprising a sample material capture substrate, wherein each sample material collector corresponds to one sample region of the plurality of sample regions;
- a first electrode in the upper region;
- a second electrode in the lower region;
- and
- a controller coupled to the first electrode and the second electrode and configured to apply a pulsed voltage through the plurality of nanostraws of between about 1 V and 100V, a pulse width of between about 10 microseconds and 50 milliseconds for a duration of between 1 second and 300 seconds.
Dependencies are inferred from the claim text.