Abstract: Method and device (1b) for performing the optical analysis of particles (2) contained in suspension in a fluid (3) arranged inside a microfluidic device (4) which maintains it at a temperature significantly lower than the ambient temperature; the formation of humidity on the outer surface (8) of the cover of the microfluidic device is avoided by applying a thermal flow (P) which determines an increase in the temperature of the outer surface (8) of the cover to above the condensation temperature (Td), or reduction in the ambient temperature (and/or humidity) in the vicinity of the cover (8), so as to bring the condensation temperature (Td) (dew point) to below the temperature of the surface (8) of the cover determined by the internal operating temperature.

Abstract: There is disclosed a method of generating a massively parallel sequencing library comprising the steps of: a) providing a primary WGA DNA library (pWGAlib), including fragments comprising a WGA library universal sequence adapter; b) performing a single PCR cycle on the pWGAlib using a first primer (1PR) comprising from 5? to 3? a first sequencing adapter (1PR5SA) and a first primer 3? section (1PR3S) hybridizing to the reverse complementary of the WGA library universal sequence adapter; c) performing a single PCR cycle on the on the product of step b) using a second primer (2PR) comprising from 5? to 3? a second sequencing adapter (2PR5SA) different from the 1PR5SA, and a second primer 3? section (2PR3S) hybridizing to the WGA library universal sequence adapter reverse complementary; d) amplifying by PCR the product of step c) using a third primer comprising the 1PR5SA and a fourth primer comprising 2PR5SA.

Abstract: A microfluidic device for the isolation of particles of at least one specific type of a sample is described. The microfluidic device comprises: a first inlet adapted to receive a sample comprising the particles of the specific type, at least part of a separation group comprising a separation unit, which comprises a main chamber and a recovery chamber and being adapted to receive the sample and to transfer at least part of the particles of the specific type from the main chamber to the recovery chamber in a selective manner with respect to further particles of the sample, a first outlet which is designed to allow the particles of the specific type to be collected outside of the device and a second outlet adapted to allow at least a portion of the sample to flow out of the main chamber and out of the microfluidic device.

Abstract: There is disclosed a method of generating a massively parallel sequencing library comprising the steps of: a) providing a primary WGA DNA library (pWGAlib), including fragments comprising a WGA library universal sequence adaptor; b) re-amplifying the primary WGA DNA library using at least one first primer (1PR) and at least one second primer (2PR); the at least one first primer (1PR) comprising from 5? to 3? at least one first sequencing adaptor (1PR5SA), at least one first sequencing barcode (1PR5BC) and a first primer 3? section (1PR3S) hybridizing to either the WGA library universal sequence adaptor or its reverse complementary; the at least one second primer (2PR) comprising from 5? to 3? at least one second sequencing adaptor (2PR5SA) different from the at least one first sequencing adaptor (1PR5SA), and a second primer 3? section (2PR3S) hybridizing to either the WGA library universal sequence adaptor or its reverse complementary.

Abstract: A method for analyzing loss-of-heterozygosity (LoH) in at least one sample comprising genomic DNA can include: a. providing the sample comprising genomic DNA; b. carrying out a deterministic restriction-site whole genome amplification (DRS-WGA) of said genomic DNA; c. preparing a massively parallel sequencing library from the product of said DRS-WGA; d. carrying out low-pass whole genome sequencing at a mean coverage depth of <1 on said massively parallel sequencing library; e. aligning the reads obtained in step d. on a reference genome for said at least one sample; f. extracting the allelic content at a plurality of loci, wherein said plurality of loci comprises polymorphic loci and/or heterozygous loci; g. assigning an LoH score to at least one genomic window of said reference genome for said at least one sample as a function of the number of loci with at least two different alleles in said plurality of loci.

Abstract: There is disclosed a method for whole genome amplification and analysis of multiple target molecules in a biological sample including genomic DNA and target molecules comprising the steps of contacting the biological sample with at least one binding agent, directed to at least one of the target molecules, conjugated with a tagged oligonucleotide, which comprises a binding-agent barcode sequence (BAB) and a unique molecular identifier sequence (UMI); carrying out a separating step to selectively remove unbound binding agent thus obtaining a labeled biological sample; simultaneously carrying out on the labeled biological sample a whole genome amplification and an amplification of the tagged oligonucleotide; preparing a massively parallel sequencing library from the amplified tagged oligonucleotide; sequencing the massively parallel sequencing library; retrieving the sequences of the BAB and UMI from each sequencing read; counting the number of distinct UMI for each binding agent.

Abstract: Methods and apparatus for the selection or processing of particles sensitive to the application of an external stimulus to rupture/lysis at least one selected particle or the fusion of first and second selected particles are disclosed herein. Particles are organized using a first field of force by selectively energizing electrodes of an array of selectable electrodes having dimensions comparable to or smaller than those of the particles. A first configuration of stresses is applied to the electrodes; and then a second configuration of stresses is applied to the electrodes, so as to create a second field of force, located substantially close to at least one selected particle to be lysated or to a pair of first and second particles to be fused and such as to produce the application of a stimulus suited to produce their lysis or fusion.

Abstract: Method and device (1b) for performing the optical analysis of particles (2) contained in suspension in a fluid (3) arranged inside a microfluidic device (4) which maintains it at a temperature significantly lower than the ambient temperature; the formation of humidity on the outer surface (8) of the cover of the microfluidic device is avoided by applying a thermal flow (P) which determines an increase in the temperature of the outer surface (8) of the cover to above the condensation temperature (Td), or reduction in the ambient temperature (and/or humidity) in the vicinity of the cover (8), so as to bring the condensation temperature (Td) (dew point) to below the temperature of the surface (8) of the cover determined by the internal operating temperature.

Abstract: The present invention relates to a method for the diagnosis of tumoural conditions and/or of the corresponding state of advance, wherein a sample from a patient comprising at least one tumour cell is obtained. According to the invention, a purified specimen of the at least one tumour cell is obtained by individually selecting and isolating single cells in a microfluidic device the purified specimen having a purity of at least 90%. On the purified specimen thus obtained there is subsequently performed a molecular analysis such as to highlight a characteristic thereof suited to enabling diagnosis.

Abstract: There is disclosed a method of generating a massively parallel sequencing library comprising the steps of: a) providing a primary WGA DNA library (pWGAlib), including fragments comprising a WGA library universal sequence adapter; b) performing a single PCR cycle on the pWGAlib using a first primer (1PR) comprising from 5? to 3? a first sequencing adapter (1PR5SA) and a first primer 3? section (1PR3S) hybridizing to the reverse complementary of the WGA library universal sequence adapter; c) performing a single PCR cycle on the on the product of step b) using a second primer (2PR) comprising from 5? to 3? a second sequencing adapter (2PR5SA) different from the 1PR5SA, and a second primer 3? section (2PR3S) hybridizing to the WGA library universal sequence adapter reverse complementary; d) amplifying by PCR the product of step c) using a third primer comprising the 1PR5SA and a fourth primer comprising 2PR5SA.

Abstract: There is disclosed a method of generating a massively parallel sequencing library comprising the steps of: a) providing a primary WGA DNA library (pWGAlib), including fragments comprising a WGA library universal sequence adaptor; b) re-amplifying the primary WGA DNA library using at least one first primer (1PR) and at least one second primer (2PR); the at least one first primer (1PR) comprising from 5? to 3? at least one first sequencing adaptor (1PR5SA), at least one first sequencing barcode (1PR5BC) and a first primer 3? section (1PR3S) hybridizing to either the WGA library universal sequence adaptor or its reverse complementary; the at least one second primer (2PR) comprising from 5? to 3? at least one second sequencing adaptor (2PR5SA) different from the at least one first sequencing adaptor (1PR5SA), and a second primer 3? section (2PR3S) hybridizing to either the WGA library universal sequence adaptor or its reverse complementary.

Abstract: The methods herein can allow for the optimised selection and isolation, within a respective population, of elements of interest and/or utility for a series of subsequent operations. A method can include the phases of: a) identifying, for each particle, at least one of a plurality of characteristic parameters; b) selecting the particles of interest, comparing for each of these the at least one parameter with a respective reference parameter. It can furthermore include the phases of c) storing, for each of the particles, the at least one parameter identified; d) processing the value of a function of the stored parameter, associating the function with a criterion for selection of the particles of interest chosen from a group of possible selection criteria; e) establishing for each particle a threshold criterion to be used as reference parameter. The threshold criterion is established on a time by time basis according to the result of the above processing.

Abstract: A microfluidic device for the isolation of particles of at least one specific type of a sample is described. The microfluidic device comprises: a first inlet adapted to receive a sample comprising the particles of the specific type, at least part of a separation group comprising a separation unit, which comprises a main chamber and a recovery chamber and being adapted to receive the sample and to transfer at least part of the particles of the specific type from the main chamber to the recovery chamber in a selective manner with respect to further particles of the sample, a first outlet which is designed to allow the particles of the specific type to be collected outside of the device and a second outlet adapted to allow at least a portion of the sample to flow out of the main chamber and out of the microfluidic device.

Abstract: A method for determining the integrity of the genome of a sample and/or the quality of a library of DNA sequences obtained by deterministic restriction site whole genome amplification can include (a) amplifying the library of DNA sequences to produce first, second, and third PCR products each of a different size from 50 bp to 1000 bp, by PCR using at least one first primer pair, one second primer pair and one third primer pair, the primer pairs each hybridizing to a DNA sequence of the library having a length from 1000 bp to 5000 bp and corresponding to a sequence of the genome located respectively on a first, second and third chromosome arm; (b) detecting the first, second and third PCR products; (c) correlating the presence of the first, second and third PCR products with the integrity of the genome of the sample and/or the quality of the library.

Abstract: The present invention relates to a method for the diagnosis of tumoural conditions and/or of the corresponding state of advance, wherein a sample from a patient comprising at least one tumour cell is obtained. According to the invention, a purified specimen of the at least one tumour cell is obtained by individually selecting and isolating single cells in a microfluidic device the purified specimen having a purity of at least 90%. On the purified specimen thus obtained there is subsequently performed a molecular analysis such as to highlight a characteristic thereof suited to enabling diagnosis.

Abstract: A method for determining the integrity of the genome of a sample and/or the quality of a library of DNA sequences obtained by deterministic restriction site whole genome amplification can include (a) amplifying the library of DNA sequences to produce first, second, and third PCR products each of a different size from 50 bp to 1000 bp, by PCR using at least one first primer pair, one second primer pair and one third primer pair, the primer pairs each hybridizing to a DNA sequence of the library having a length from 1000 bp to 5000 bp and corresponding to a sequence of the genome located respectively on a first, second and third chromosome arm; (b) detecting the first, second and third PCR products; (c) correlating the presence of the first, second and third PCR products with the integrity of the genome of the sample and/or the quality of the library.

Abstract: Method for the treatment of a biological sample comprising at least one cell and a liquid component; according to the method, a force is applied to the sample inserted in an inner chamber of a hollow device towards a filter which has pores with diameters from 2 nm to 1 ?m, so that at least part of the liquid component passes through the filter and the cell remains in the inner chamber, thus obtaining a concentrated sample; the filter has a surface facing the inner chamber of less than 12.6 mm2.

Abstract: Methods and apparatus for the selection or processing of particles sensitive to the application of an external stimulus to rupture/lysis at least one selected particle or the fusion of first and second selected particles are disclosed herein. Particles are organized using a first field of force by selectively energizing electrodes of an array of selectable electrodes having dimensions comparable to or smaller than those of the particles. A first configuration of stresses is applied to the electrodes; and then a second configuration of stresses is applied to the electrodes, so as to create a second field of force, located substantially close to at least one selected particle to be lysated or to a pair of first and second particles to be fused and such as to produce the application of a stimulus suited to produce their lysis or fusion.

Abstract: The present invention relates to a method for the diagnosis of tumoural conditions and/or of the corresponding state of advance, wherein a specimen of an organic fluid taken from the patient and having a high probability of containing at least one circulating tumor cell (CTC) or a disseminated tumor cell (CTD) is enriched in a population of CTCs or CTDs. According to the invention, at least one type of CTCs or CTDs is isolated by selecting individually single cells in a microfluidic device so to constitute a diagnostic specimen having a purity of at least 90%. On the specimen thus obtained there is subsequently performed a molecular analysis such as to highlight a characteristic thereof suited to enabling diagnosis.

Abstract: Methods and apparatus for the selection or processing of particles sensitive to the application of an external stimulus to rupture/lysis at least one selected particle or the fusion of first and second selected particles are disclosed herein. Particles are organized using a first field of force by selectively energizing electrodes of an array of selectable electrodes having dimensions comparable to or smaller than those of the particles. A first configuration of stresses is applied to the electrodes; and then a second configuration of stresses is applied to the electrodes, so as to create a second field of force, located substantially close to at least one selected particle to be lysated or to a pair of first and second particles to be fused and such as to produce the application of a stimulus suited to produce their lysis or fusion.