Provided herein are methods and systems for processing preserved tissue and/or dissociating tissue to release cells, nuclei and/or other subcellular organelles. Instruments comprise an interface for engaging cartridges through fluidic lines to introduce or remove liquids from a processing chamber of a cartridge. Systems also include reagent subsystems configured to prevent liquid spills and securely engage fittings between reagent containers and fluidic lines. Systems also include real-time feedback on the pressure on a grinding rotor in a processing chamber of a cartridge.
A cartridge for dissociating tissue, comprising: a processing chamber comprising a stator, a side wall, a top orifice, and a first processing chamber port positioned in the side wall; and a grinder assembly comprising a plunger comprising a rotor, a grinder assembly slidably positioned in the processing chamber through the top orifice; wherein: the stator comprises a plurality of teeth arranged in a spaced-apart array of rings; and the rotor comprises one or more central teeth and a plurality of teeth arranged in a spaced-apart array of rings, wherein one ring of teeth is positioned at or substantially at a circumference of the rotor; wherein the rings in the stator and the rings in the rotor are positioned such that when the rotor contacts the stator, rings of teeth in the stator mesh with the one or more central teeth and rings of teeth in the rotor
A system, methods, and apparatus are described to collect and prepare single cells, nuclei, subcellular components, and biomolecules from specimens including tissues and in some embodiments use the single cells to form organoids or microtissues. The system can perform enzymatic and/or physical disruption of the tissue to dissociate it into single-cells and then use a hanging droplet method to form organoids or microtissues.
Systems, methods, and apparatus are described to collect and prepare cells, nuclei, subcellular components, and biomolecules from specimens including tissues. Some of the systems provided herein comprise an instrument and cartridge. The instrument can comprise at least one cartridge interface configured to engage at least one cartridge. In some cases, the cartridge comprises a functional unit (e.g., a processing chamber comprising a grinder assembly) and at least one cartridge bay or vessel bay, wherein the cartridge bay or vessel bay comprises attachment features permitting at least one functional unit to engage with the cartridge. The functional unit or units can include at least one of a filtration unit, a magnetic processing unit, a dissociation chamber, a processing chamber, an output unit, a flowcell unit, a tangential flow filtration unit, and a waste unit.
G01N 35/02 - Analyse automatique non limitée à des procédés ou à des matériaux spécifiés dans un seul des groupes Manipulation de matériaux à cet effet en utilisant une série de récipients à échantillons déplacés par un transporteur passant devant un ou plusieurs postes de traitement ou d'analyse
C12M 1/00 - Appareillage pour l'enzymologie ou la microbiologie
G01N 35/00 - Analyse automatique non limitée à des procédés ou à des matériaux spécifiés dans un seul des groupes Manipulation de matériaux à cet effet
This disclosure provides methods for producing a sample of subcellular organelles, particularly nuclei, from a tissue. In some embodiments, this disclosure provides a method of processing a tissue sample involves performing enzymatic/chemical disruption of tissue in a chamber to produce disrupted tissue comprising released cells and/or nuclei and debris; separating the released cells and/or nuclei from the debris therein; and moving the released cells and/or nuclei. In some instances, the method comprises mechanical disruption of the tissue sample.
A system, methods, and apparatus are described to collect and prepare single cells, nuclei, subcellular components, and biomolecules from specimens including tissues and in some embodiments use the single cells to form organoids or microtissues. The system can perform enzymatic and/or physical disruption of the tissue to dissociate it into single-cells and then use a hanging droplet method to form organoids or microtissues.
A cartridge for dissociating tissue, comprising: a processing chamber comprising a stator, a side wall, a top orifice, and a first processing chamber port positioned in the side wall; and a grinder assembly comprising a plunger comprising a rotor, a grinder assembly slidably positioned in the processing chamber through the top orifice; wherein: the stator comprises a plurality of teeth arranged in a spaced-apart array of rings; and the rotor comprises one or more central teeth and a plurality of teeth arranged in a spaced-apart array of rings, wherein one ring of teeth is positioned at or substantially at a circumference of the rotor; wherein the rings in the stator and the rings in the rotor are positioned such that when the rotor contacts the stator, rings of teeth in the stator mesh with the one or more central teeth and rings of teeth in the rotor
A system, methods, and apparatus are described to collect and prepare cells, nuclei, subcellular components, and biomolecules from specimens including FFPE and OCT preserved tissues. The system can perform deparaffinization, rehydration, enzymatic and/or chemical and physical disruption of the FFPE tissue, or residue removal of the OCT tissue, to dissociate it into a single cell or nuclei suspension.
A system, methods, and apparatus are described to collect and prepare single cells, nuclei, subcellular components, and biomolecules from specimens including tissues and in some embodiments use the single cells to form organoids or microtissues. The system can perform enzymatic and/or physical disruption of the tissue to dissociate it into single-cells and then use a hanging droplet method to form organoids or microtissues.
This disclosure provides methods for producing a sample of subcellular organelles, particularly nuclei, from a tissue. In some embodiments, this disclosure provides a method of processing a tissue sample involves performing enzymatic/chemical disruption of tissue in a chamber to produce disrupted tissue comprising released cells and/or nuclei and debris; separating the released cells and/or nuclei from the debris therein; and moving the released cells and/or nuclei. In some instances, the method comprises mechanical disruption of the tissue sample.
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.
A system, methods, and apparatus are described to collect and prepare cells, nuclei, subcellular components, and biomolecules from specimens including FFPE and OCT preserved tissues. The system can perform deparaffinization, rehydration, enzymatic and/or chemical and physical disruption of the FFPE tissue, or residue removal of the OCT tissue, to dissociate it into a single cell or nuclei suspension.
A system, methods, and apparatus are described to collect and prepare single cells, nuclei, subcellular components, and biomolecules from specimens including tissues and in some embodiments use the single cells to form organoids or microtissues. The system can perform enzymatic and/or physical disruption of the tissue to dissociate it into single-cells and then use a hanging droplet method to form organoids or microtissues.
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.
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.
C12M 3/00 - Appareillage pour la culture de tissus, de cellules humaines, animales ou végétales, ou de virus
C12M 3/06 - Appareillage pour la culture de tissus, de cellules humaines, animales ou végétales, ou de virus avec des moyens de filtration, d'ultrafiltration, d'osmose inverse ou de dialyse
C12M 3/08 - Appareils pour la désagrégation des tissus
C12N 1/08 - Réduction de la teneur en acide nucléique
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