Cell ultrasonication

Resuspend in 70% ethanol, disaggregate in French pressure cell, ultrasonicate and wet sieve. 

Fractional sonication - phase partition. Twenty ml of the lower phase B3 and 10 ml of fresh upper phase was sonicated 4 x 30 s with 2 min resting intervals under continuous cooling using a Vibra-cell ultrasonic processor, Model VC 500, equipped with a 3/4 inch "High Gain" horn. The output was set at 7 with 201 duty pulses. The phases were separated by low speed centrifugation and the upper phase was removed and stored separately. 

Wu, J., Lin, L., and Chau, F. 2001. Ultrasound-assisted extraction of ginseng saponins from ginseng roots and cultured ginseng cells. Ultrason. Sonochem. 8 347-352. 

There are several other means of disrupting cells ultrasonication, freezing and thawing, hypotonic treatment, etc. These procedures may be attempted when coaxial homogenizers and blendors prove ineffective. Unfortunately, they are generally injurious to mammalian subcellular structures. 

Two major sources of ultrasound are employed, namely ultrasonic baths and ultrasonic immersion hom probes [79, 71]- The fonuer consists of fixed-frequency transducers beneath the exterior of the bath unit filled with water in which the electrochemical cell is then fixed. Alternatively, the metal bath is coated and directly employed as electrochemical cell, but m both cases the results strongly depend on the position and design of the set-up. The ultrasonic horn transducer, on the other hand, is a transducer provided with an electrically conducting tip (often Ti6A14V), which is inuuersed in a three-electrode thenuostatted cell to a depth of 1-2 cm directly facing the electrode surface. 

Fig. 14 Effect of the molecular weight of tamarind seed xyloglucan depolymerized by ( ) 7-irradiation, ( ) ultrasonication, and ( ) endo-glucanase treatment on the production of various cytokines (Tumor necrosis factor a, TNE-a Interleukin 8, IL-8 Interleukin 10, IL-10 and Interleukin 12, IL-12) in HaCaT cells (Immortalized keratinocytes line) [301]...

Doulah MS (1977) Mechanism of disintegration of biological cells in ultrasonic cavitation. Biotech Bioeng 19 649-660... 

Fig. 4.1 (a) Electrochemical cell dipped in ultrasonic bath, (b) Klima sonoelectrochemical cell (Reprinted from [35]. Copyright (1999) with the permission from Elsevier)... 

It should also be important to highlight that, in addition to these two more popular experimental set-up, the early electrochemical glass cell dipped in ultrasonic bath configuration is also being used with satisfactory results [8, 32],... 

He et al. (2002) used an off-line HPLC/CE method to map cancer cell extracts. Frozen ovarian cancer cells (containing 107 cells) were reconstituted in 300 pL of deionized water and placed in an ultrasonic bath to lyse the cells. Then the suspension was centrifuged and the solubilized proteins were collected for HPLC fractionation. The HPLC separation was carried out on an instrument equipped with a RP C-4 column, 250 mm x 4.6 mm, packed with 5-pm spherical silica particles. Extracted proteins were dissolved in 300 pL of DI water, and lOOpL was injected onto the column at a flow rate of 1 mL/min. Buffer A was 0.1% TEA in water and buffer B was 0.1% TFA in acetonitrile. A two-step gradient, 15-30% B in 15 min followed by 30-70% B in 105 min, was used. The column effluent was sampled every minute into a 96-well microtiter plate with the aid of an automatic fraction collector. After collection, the fractions were dried at room temperature under vacuum. The sample in each well was reconstituted before the CE analysis with 10 pL deionized water. The... 

Large-scale ultrasonic irradiation is extant technology. Liquid processing rates of 200 liters/minute are routinely accessible from a variety of modular, in-line designs with acoustic power of several kW per unit (83). The industrial uses of these units include (1) degassing of liquids, (2) dispersion of solids into liquids, (3) emulsification of immiscible liquids, and (4) large-scale cell disruption (74). While these units are of limited use for most laboratory research, they are of potential importance in eventual industrial application of sonochemical reactions. 

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