For analyzing a sample containing particles of at least two categories, such as a sample containing blood cells, a particle counter subject to a detection limit is coupled with an analyzer capable of discerning particle number ratios, such as a visual analyzer, and a processor. A first category of particles can be present beyond detection range limits while a second category of particles is present within respective detection range limits. The concentration of the second category of particles is determined by the particle counter. A ratio of counts of the first category to the second category is determined on the analyzer. The concentration of particles in the first category is calculated on the processor based on the ratio and the count or concentration of particles in the second category.

The present disclosure relates to apparatus, systems, compositions, and methods for analyzing a sample containing particles. In some aspects the system comprises an analyzer which may be a visual analyzer. In one aspect, this disclosure relates to a particle imaging system comprising a flowcell through which a sample containing particles is caused to flow, and a high optical resolution imaging device which captures images for image analysis of samples. Other compositions, methods and features of this disclosure are disclosed herein.

A technique is presented for aligning, in a desired region within a flow chamber of a flow cell, a non-spherical biological entity carried in a sample. The flow chamber has a rectangular cross-section. A bottom flow input module, a top flow input module and a sample input module provide a viscoelastic first fluid, a second viscoelastic fluid, and the sample, respectively, to the flow chamber. The first and the second viscoelastic fluids laminarly flow along a bottom and a top wall of the flow chamber and the sample laminarly flows sandwiched between them. By controlling rate of flow of the first and/or the second viscoelastic fluids the sample flow, and thus the non-spherical biological entity, is focused in the desired region. A gradient of sheer within the sample flow set up due to the first and second viscoelastic fluids orients the non-spherical biological entity in the desired region.

Aspects and embodiments of the instant disclosure provide a particle and/or intracellular organelle alignment agent for a particle analyzer used to analyze particles contained in a sample. An exemplary particle and/or intracellular organelle alignment agent includes an aqueous solution, a viscosity modifier, and/or a buffer. Embodiments also encompass systems, compositions, and methods for analyzing a sample containing particles. Parrticles such as blood cells can be categorized and counted by a digital image processor. A digital microscope camera can be directed, for example using certain focusing techniques, into a flowcell defining a symmetrically narrowing flowpath in which the sample stream flows in a ribbon flattened by flow and viscosity parameters between layers of sheath fluid. Blood cell images can be collected and analyzed using dynamic range extension processes and systems.

Apparatus for sorting and orienting sperm cells has a pair or walls in confronting relationship forming a flow chamber having inlet, a downstream outlet, and intermediate detector region. The inlet receives first and second spaced apart streams of input fluid and a third stream of sample fluid containing the cells to be sorted. The first and second streams have respective flow rates relative to third stream, such that the third stream is constricted forming a relatively narrow sample stream, so that the cells are oriented parallel to the walls. A detector detects desired cells and a sorter downstream of the detector for sorting the desired cells from the stream.

Disclosed herein is a method for detecting cell aggregates of biological cells using a quantitative phase-contrast microscope and a device for detecting cell aggregates of biological cells using said method. The method comprises preparing a suspension comprising biological cells from a sample. A flow of the suspension is generated along a microfluidic channel to viscoelastically and/or hydrodynamically focus cell aggregates in the suspension in a focal plane of the quantitative phase-contrast microscope. One or more phase shift images of the biological cells in the suspension are taken using the quantitative phase-contrast microscope. Cell aggregates in the one or more phase shift images are identified. The sample is a whole blood sample or a blood fraction sample and identifying cell aggregates in the one or more phase shift images comprises identifying platelet aggregates in the one or more phase shift images.

The present disclosure relates to apparatus, systems, compositions, and methods for analyzing a sample containing particles. In some aspects the system comprises an analyzer which may be a visual analyzer. In one aspect, this disclosure relates to a particle imaging system comprising a flowcell through which a sample containing particles is caused to flow, and a high optical resolution imaging device which captures images for image analysis of samples. Other compositions, methods and features of this disclosure are disclosed herein.

Disclosed herein is a method for detecting cell aggregates of biological cells using a quantitative phase-contrast microscope, a device for detecting cell aggregates of biological cells, a method for detecting cellular and/or molecular biological objects using a quantitative phase-contrast microscope and a device for detecting cellular and/or molecular biological objects. The method for detecting cell aggregates of biological cells using a quantitative phase-contrast microscope comprises preparing a suspension, the suspension comprising a viscoelastic fluid and biological cells from a sample, wherein the viscoelastic fluid comprises a shear-thinning polymer having a molecular weight between 2 MDa and 10 MDa and wherein a mass fraction of the shear-thinning polymer in the suspension is less than 0.2%. A flow of the suspension is generated along a microfluidic channel to viscoelastically focus cell aggregates in the suspension in a focal plane of the quantitative phase-contrast microscope. One or more phase shift images of the biological cells in the suspension are taken using the quantitative phase-contrast microscope and cell aggregates are identified in the one or more phase shift images.

This invention provides microfabricated devices and methods for detecting, analyzing and sorting biological materials and particles. Droplets containing the particles are provided in an extrusion fluid, passed through a detection region, and then directed into a branch channel according to predetermined characteristics. For example, cells or viral particles contained in droplets of aqueous solvent are flowed past a detector in the nonpolar extrusion fluid decane, and routed into a selected branch channel for subsequent analysis or use.

The present disclosure relates to apparatus, systems, compositions, and methods for analyzing a sample containing particles. In some aspects the system comprises an analyzer which may be a visual analyzer. In one aspect, this disclosure relates to a particle imaging system comprising a flowcell through which a sample containing particles is caused to flow, and a high optical resolution imaging device which captures images for image analysis of samples. Other compositions, methods and features of this disclosure are disclosed herein.