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Novel millimeter-wave-based method for in situ cell isolation and other applications.

Scientific reports (2018-10-05)
Barney Boyce, Natalia Samsonova
ABSTRACT

As an alternative to laser-based methods, we developed a novel in situ cell isolation method and instrument based on local water absorption of millimeter wave (MMW) radiation that occurs in cellular material and nearby culture medium while the cultureware materials (plastic and glass) are transparent to MMW frequencies. Unwanted cells within cell population are targeted with MMWs in order to kill them by overheating. The instrument rapidly (within 2-3 seconds) heats a cell culture area of about 500 µm in diameter to 50 °C using a low-power W-band (94 GHz) MMW source. Heated cells in the area detach from the substrate and can be removed by a media change leaving a bare spot. Hence we named the instrument "CellEraser". Quick, local and non-contact heating with sharp boundaries of the heated area allows elimination of the unwanted cells without affecting the neighboring cells. The instrument is implemented as a compact microscope attachment and the selective hyperthermic treatment can be done manually or in an automated mode. Mammalian cells heated even momentarily above 50 °C will not survive. This "temperature of no return" does not compromise cellular membranes nor does it denature proteins. Using the CellEraser instrument we found that the key event that determines the fate of a cell at elevated temperatures is whether or not the selectivity of its nucleus is compromised. If a cell nucleus becomes "leaky" allowing normally excluded (cytoplasmic) proteins in and normally nuclear-localized proteins out, that cell is destined to die. Quick heating by MMWs to higher temperatures (70 °C) denatures cellular proteins but the cells are not able to detach from the substrate - instead they undergo a phenomenon we called "thermofixation": such cells look similar to cells fixed with common chemical fixatives. They remain flat and are not washable from the substrate. Interestingly, their membranes become permeable to DNA dyes and even to antibodies. Thermofixation allows the use of western blot antibodies for immunofluorescence imaging.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Etoposide, synthetic, 98.0-105.0%, powder
Sigma-Aldrich
U2OS LMNB1-TUBA1B-ACTB

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