The present invention is directed to methods and apparatus for and products from disrupting, removing intracellular proteins, enzymes and nucleic acids, spray drying, lipid extraction, and making nanoparticles of Saccharomyces cerevisiae (yeast) cell wall followed by acid and/or enzymatic hydrolysis to produce Beta (β)-glucans, chitin and mannans (mannoproteins). The process and apparatus feature critical, supercritical, or near critical fluids for disruption of yeast and making yeast cell wall nanoparticles. The product materials retain full activity and are devoid of residual processing chemicals such as solvents, salts, or surfactants.

In one example in accordance with the present disclosure, a cellular analytic system is described. The cellular analytic system includes an analytic device. The analytic device includes a chamber to receive a cell to be analyzed. At least one lysing element agitates the cell and at least one sensor detects a change in the cell based on an agitation of the cell. The cellular analytic system also includes a controller to determine a rupture threshold of the cell based on parameters of the agitation when a cell membrane ruptures.

Devices and methods are provided for electrically lysing cells and releasing macromolecules from the cells. A microfluidic device is provided that includes a planar channel having a thickness on a submillimeter scale, and including electrodes on its upper and lower inner surfaces. After filling the channel with a liquid, such that the channel contains cells within the liquid, a series of voltage pulses of alternating polarity are applied between the channel electrodes, where the amplitude of the voltage pulses and a pulse width of the voltage pulses are effective for causing irreversible electroporation of the cells. The channel is configured to possess thermal properties such that the application of the voltage produces a rapid temperature rise as a result of Joule heating for releasing the macromolecules from the electroplated cells. The channel may also include an internal filter for capturing and concentrating the cells prior to electrical processing.

Magnetic beads having cell components of interest are translated between a sequence of processing wells in a tray without need for pipetting. The circular tray contains one or more sequences of wells each interconnected by a respective channel. The tray is rotated about a central axis and a magnet, an agitator, and a heater provided external to the tray enable magnetic bead translation, mixing, and incubation, respectively. The magnet proximate a well forms a cluster of beads. Manipulation of the tray in rotation and elevation results in translation of the cluster from one well, through a channel, and into an adjacent well. The well containing a cluster may be rotationally positioned in front of the agitator, the agitator extended into contact with the well, followed by mechanical agitation. The heater, disposed beneath the tray, may accept a well lowered thereto for selective heating.

In one example in accordance with the present disclosure, a cellular analytic system is described. The cellular analytic system includes a series of analytic devices. Each analytic device includes 1) a separator to separate a cellular particle from a surrounding fluid, 2) an analyzer coupled to a first outlet of the separator to analyze the surrounding fluid, and 3) at least one lysing device coupled to at least a second outlet of the separator to rupture a membrane of the cellular particle. An outlet of the lysing device is fluidly coupled to a separator of a downstream analytic device.

The present disclosure provides improved methods for bead beating and a bead beating system useful therefor. The present disclosure further provides methods of using the bead beating system to extract nucleic acids from cells containing the nucleic acids.

A method for preparing a sample by utilizing a shearing force in the presence of a size stabilizer to break apart the sample to obtain nucleic acid molecules in a usable size range. Once nucleic acid molecules are obtained, magnetic nanoparticles are used to concentrate and clean the nucleic acid molecules for further testing.

Disrupting a biological cell includes freezing, boiling or perhaps alternately freezing and boiling material containing the biological cell using a thermoelectric cell with a working face, and a base face whereof is contiguous with a heat source/sink at a substantially constant temperature. Apparatus for the disruption process includes a peltier cell, a base face, which is flexibly attached to a heat source/sink held at a constant temperature, and a working face contiguous with a reaction vessel or holder thereof. Reversal of the voltage in the peltier cell enables the working face alternately to reach below freezing and above boiling temperatures, and/or with use of a resistive wire on the vessel or holder for heating, with the TEC used purely for cooling. The materials of the base face tend to inhibit disintegration of the peltier cell brought about by expansion and contraction by heat.

The present invention provides an apparatus and a method for a lysis procedure, in particular for an automated and/or controlled lysis procedure of a sample, in particular a biological sample. The apparatus comprises at least one rotation disc (31), at least one vial holder (90) which is configured to receive a vial (100), wherein the vial holder (90) is arranged on the disc (31), at least one driving device (20) which is configured to rotate the disc (31) and the vial holder (90), at least one heating device (60) which is configured to heat the sample to a determined incubation temperature, and—at least one control device (70) which is configured to control the driving device (20) and/or the heating device (60) by means of a timing and/or step control, and/or—at least one transmitting device (80) for inductive coupling for energy and signal transmission, which is configured to transmit the energy for heating to the heating device (60), and/or—wherein the driving device (20) is configured to rotate the disc (31) in a first direction (A1) and/or with a first speed, and to rotate the vial holder (90) in a second direction (A3) and/or with a second speed. The apparatus and the method are adapted for an (automated) lysis procedure, wherein the lysis can be carried out in a safe, efficient and effective manner.

A system, methods, and apparatus are described to collect and prepare single cells, nuclei, subcellular components, and biomolecules from specimens including tissues. The system can perform enzymatic and/or physical disruption of the tissue to dissociate it into single-cells or nuclei in suspension or subcellular components including nucleic acids. In some embodiments, the titer of dissociated cells is monitored at intervals and the viability determined. In some embodiments, the processing is adjusted according to the measurements of the titer and viability. In some embodiments, the single-cells or nuclei in suspension are washed and resuspended in the buffer or media of choice. In some embodiments, the conditions are chosen to produce nuclei. In other embodiments, the single-cells or nuclei are purified by affinity paramagnetic bead processing. In some embodiments, matched bulk nucleic acid to the single-cells is produced. In other embodiments, single-cell libraries, or nuclei libraries, or matched bulk libraries, or bulk libraries are produced. The single cells or nuclei can then be further processed by FACS, DNA sequencing, mass spectrometry, fluorescence, or other methods. In other embodiments, the tissue processing is integrated with an analytical system to produce a sample-to-answer system such as a tissue-to-genomics system.