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.
Claims14
A method of nondestructive sampling of intracellular sample material from within a cell, the method comprising:
- applying a voltage between an upper electrode and a lower electrode through a nanostraw to open one or more pores in a portion of the cell membrane extending over an opening of the nanostraw;
- capturing a sample material released from within the cell and into the nanostraw in a sample collector beneath the nanostraw;
- and
- stopping the application of voltage between the upper and lower electrodes and allowing the cell membrane to recover before more than 15% of the sample material within the cell is released.
A method of nondestructive sampling of intracellular sample material from within a cell at multiple time points, the method comprising:
- applying a voltage of between 1 and 100 V between an upper electrode and a lower electrode through a nanostraw to open one or more pores in a portion of the cell membrane extending over an opening of the nanostraw;
- capturing a sample material released from within the cell and into the nanostraw in a sample collector beneath the nanostraw, wherein capturing comprises immobilizing the sample material onto a capture substrate;
- stopping the application of voltage between the upper and lower electrodes and allowing the cell membrane to recover before more than 15% of the sample material within the cell is released;
- and
- allowing the cell to recover for a minimum recovery time of at least 1 hour before reapplying the voltage and capturing additional sample material.
Dependencies are inferred from the claim text.