Freezing and Archiving Cells on Microfluidic Devices
A microfluidic device and cell technology, applied in enzymology/microbiology devices, human or animal body preservation, biochemical cleaning devices, etc., can solve problems such as inability to interact with microfluidic devices, occupying freezing space, etc.
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example 1
[0194] Example 1 - Freezing of 1F5 hybridoma cells (ATCC HB-9645) on a microfluidic device.
[0195] The inventors of the present application introduced biological cells grown in standard culture plates into a microfluidic device at 36°C. Cells were then manipulated and isolated in individual isolation regions in the microfluidic device (still at 36°C). A 15% DMSO solution (in media) was poured into the microfluidic device, and the device was incubated at 36°C for 30 minutes without perfusion. The microfluidic device was then cooled to 0°C at a controlled rate of 1.8°C / min. It was observed that during freezing, the isolated cells continued to move, but at a slower rate as the temperature dropped. The device was placed in a Styrofoam box, which was then placed in a -80°C freezer overnight. The next day, the microfluidic device was removed from the freezer (and from the Styrofoam box) and allowed to warm up to room temperature (assuming the device temperature rose to room tem...
example 2
[0196] Example 2 - Freezing of prostate cancer cells on a microfluidic device.
[0197] The tissue biopsy is broken down into individual cells and introduced into the microfluidic device. Dilute 12 mL of cell suspension at 1 x 10 6 A concentration of cells / mL was input into the device, which had a total channel volume of approximately 1.0 mL, with a nominal volume close to 1000 cells at any one time. Cells were placed individually into isolation chambers using an automated penning algorithm. During cell loading and isolation, the device temperature was set at 12 °C. The microfluidic device is then placed in the freezer for sufficient time. The device was then thawed by removing it from the freezer and placed on the device (T = 12°C). Live cells in the isolation chamber were observed to be mobile using OET and could be exported for further analysis, such as sequencing.
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