Dose and location controlled drug/gene/particle delivery to individual cells by nanoelectroporation
A simple and low cost method of producing sealed arrays of laterally ordered nanochannels interconnected to microchannels of tunable size, over large surface areas, is disclosed. The method incorporates DNA combing and subsequent imprinting. Associated micro and macroscale inlets and outlets can be formed in the same process or manufactured later in low cost, non-cleanroom techniques. The techniques embrace two procedures, generating DNA nanostrands and translating these strands into nanoscale constructs via imprinting. Devices incorporating the novel arrays have a first microchannel, a second microchannel and a nanochannel that is substantially linear and which defines an axis. The nanochannel is connected at its open ends to the microchannels, which are aligned along the axis. Methods for precise dose delivery of agents into cells employing the devices in nanoelectroporation methods are also disclosed.
1. A method for transfecting cells, comprising the steps of:
providing a nanochannel electroporation device, the device comprising an array of transfection sites, each transfection site having a first microchannel, a second microchannel and a nanochannel having a first opening at the first microchannel and a second opening at the second microchannel, the nanochannel connecting and extending between the first and second microchannels;
providing, in the respective first microchannels of the array, a transfecting agent;
loading, in the respective second microchannels of the array, a cell to be transfected, the cell positioned at the second opening of the respective nanochannel; and
applying a voltage, in excess of 200 V, across the nanochannel.
2. The method of claim 1 , wherein:
applying the voltage transfects the cell with a predetermined dose of the transfecting agent.
3. The method of claim 2 , wherein:
the voltage is applied for milliseconds to transfect the cell with an error of the dose of less than 20%.
4. The method of claim 1 , wherein:
the transfecting agent is chosen from the group consisting of plasmids, biomolecules, fluorophores, dyes, RNA, DNA, and proteins.
5. The method of claim 1 , wherein:
the cell is viable after transfection.
6. The method of claim 1 , wherein the loading step is achieved by applying a centrifugal force.
7. The method of claim 1 , wherein the loading substep is achieved by optical tweezers.
8. The method of claim 2 , wherein:
a pulse number and duration of the voltage applied determines an amount of the predetermined dose transfected.
9. The method of claim 1 , wherein the nanochannels in the array of transfection sites are laterally ordered and each nanochannel defines an axis, with the first and second microchannel connected to each nanochannel aligned with the axis.
10. The method of claim 2 , wherein the transport of the transfecting agent across the nanochannel is dominated by electrophoresis occurring during the step of applying voltage across the nanochannel.