Massively parallel high throughput single-cell optoporation
Massively parallel high throughput single-cell optoperation and their uses thereof are disclosed. The disclosed method teaches the use of parallel single-cell printing technique using SU-8 membrane and nanosecond pulsed laser to espouse on single-cell with micro-dish pattern structure. The platform is able to effectively deliver different (small to large) cargo in a different cell type with high transfection efficiency and high cell viability at parallel single-cell resolution. The proposed Bio-MEMS device has potential applications in personalized and regenerative medicine.
1 . A method for printing a plurality of individual cells in parallel, said method comprising:
fabricating an SU-8 membrane material with an array of holes using a photolithography technique on a glass substrate in order to use the SU-8 membrane material as part of a cell printing platform, wherein the plurality of individual cells are cultured and seeded on the SU-8 membrane material, wherein a size of each hole in the array of holes of the SU-8 membrane material is designed such that respective individual cells of the plurality of individual cells can pass through each hole of the SU-8 membrane material to create a pattern of cells on the glass substrate;
flipping an array of microdish patterned metal on the glass substrate and placing the array of microdish patterned metal on the glass substrate, such that the array of microdish patterned metal faces the pattern of cells; and
exposing the glass substrate with a pulsed laser at a near infrared wavelength (NIR) to create photothermal vapor induced bubbles, wherein the photothermal vapor induced bubbles create pores in a cell membrane of each of the individual cells of the plurality of individual cells and each cell membrane is used to diffuse desired molecules in each of the individual cells of the plurality of individual cells.
2 . The method of claim 1 , further comprising preparing the glass substrate by: cutting a glass microslide into a square, cleaning the glass microslide with a standard piranha cleaning process, drying the glass microslide with nitrogen blow, and dehydrating the glass microslide.
3 . The method of claim 2 , wherein fabricating the SU-8 membrane material comprises spin coating SU-8 3005 on the glass substrate, patterning the SU-8 3005 , ultra-sonicating the SU-8 3005 in a developer for 1 minute to release the SU-8 membrane material from the glass substrate, cleaning the SU-8 membrane material with isopropyl alcohol (IPA) and deionized water, and drying the SU-8 membrane material using nitrogen.
4 . The method of claim 2 , further comprising peeling the SU-8 membrane material off from the glass substrate and putting the SU-8 membrane material into a petri dish wherein the petri dish was UV treated with the Su8 membrane for 2 hrs.