Concentration of Cryoprotectant in water-in-oil microdroplets for single cell vitrificaton

(cont.) Droplets with an initial concentration of IM were found to be concentrated to about 3-4M in 90s while droplets starting at 2M were concentrated to 6M in about the same time. The entire process takes place over a time scale of about one minute, fast enough to minimize exposure times but slow enough to be precisely controllable. This phenomenon is demonstrated to dynamically concentrate cryoprotectants within single cell-containing droplets. These droplets of sizes of about 30 micron diameter were concentrated to 3-4M from a starting concentration of IM in about 300s. The cells are tolerant to this concentration process and do not die when subjected to it. The process may be used in practice to innocuously concentrate cell encapsulating droplets which may then be vitrified before they are exposed to high temperatures for fatally long time scales. With appropriate characterization, the controllability of the process will allow for choosing exact cryoprotectant concentration levels used for vitrification. The demonstrated phenomenon has several other applications in cryobiology. Its controllability and speed may be used to dynamically modulate cryoprotectant concentrations in preservation protocols that require stepwise concentration or dilution. In addition, the process was found to be reversible and may thus be used for unloading cryoprotectants by controlled cooling as opposed to heating.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Mechanical Engineering.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2008

Author dc:creator

• Bajpayee, Anurag

Advisor dc:contributor.advisor

• Mehmet Toner and Gang Chen.

• Mechanical Engineering.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/46055

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/46055