Journal: Nature communications

Article Title: Inertio-elastic focusing of bioparticles in microchannels at high throughput

doi: 10.1038/ncomms5120

Figure Lengend Snippet: Inertio-elastic focusing of bioparticles based on deformability and shape ( a ) Deformation statistics of WBCs in PBS, a low-molecular weight (357 kDa) HA solution and a high-molecular weight (1,650 kDa) HA solution. The magnitude of WBC stretching is expressed in terms of aspect ratio AR = a x /a z . Scale bar, 10 µm. The error bars indicate the s.d. in the WBC aspect ratio at each flow rate. ( b ) LEF and particle trajectory analysis images of WBCs in PBS, 357 kDa HA solution and 1,650 kDa HA solution at Q = 13 ml min −1 . ( c ) Particle trajectory analysis (PTA) images of anisotropic PEG particles in 1,650 kDa HA solution at Q = 20 ml min −1 . Dashed red lines indicate channel centerline. Scale bar, 30 µm. Measurements of lateral position z and instantaneous orientation angle θ are plotted for each PEG particle in water (blue) and in the HA solution (green).

Article Snippet: HA sodium salt (357 kDa (Lifecore Biomedical) and 1650 kDa (Sigma-Aldrich)) was added to water (Sigma-Aldrich) for bead suspensions or phosphate-buffered saline (PBS) solution (Life Technologies) solution for cell suspensions and prepared using a roller mixer (Stuart, Sigma-Aldrich).

Techniques: Molecular Weight, Flow Cytometry

Journal: Journal of Extracellular Vesicles

Article Title: Fast characterisation of cell-derived extracellular vesicles by nanoparticles tracking analysis, cryo-electron microscopy, and Raman tweezers microspectroscopy

doi: 10.3402/jev.v1i0.19179

Figure Lengend Snippet: Comparison of the Raman spectra of D. discoideum growth and starvation EVs. (A) Proposed interpretation of Raman spectrum originated from vesicles released from D. discoideum in the starvation phase. This spectrum was obtained by averaging 15 different sets of vesicles, with a total accumulation time of 14 minutes, then subtraction of the PBS contribution, and smoothing by 2 adjacent pixels. (B) Variability of Raman spectra of D. discoideum vesicles during growth (a, b) as compared to starvation phase (c, d). Within the same phase, one can qualitatively distinguish at least 2 different species: during growth, species (b) is characterised by an increased amount of lipids as compared to species (a), and during starvation, species (d) contains an increased amount of nucleic acids and carotenoids as compared to species (c). The prominent spectral differences are highlighted by dashed oval curves.

Article Snippet: EVs, present in the 12,000×g pellet, are resuspended in a phosphate-buffered solution (PBS) [pH 7.4 without calcium and magnesium (Gibco)], with a 20 to 100 volume concentration factor (X), relative to the volume of the 2,000×g supernatant.

Techniques:

Journal: Journal of Extracellular Vesicles

Article Title: Fast characterisation of cell-derived extracellular vesicles by nanoparticles tracking analysis, cryo-electron microscopy, and Raman tweezers microspectroscopy

doi: 10.3402/jev.v1i0.19179

Figure Lengend Snippet: Size distribution and concentration of D. discoideum EVs samples ( Table I ), as measured by NTA. EVs were prepared from growth medium after 48 hours of cell growth, either with the usual conditions for the 12,000×g centrifugation (in Eppendorf tubes) (a), or with the 12,000×g centrifugation performed in 30 ml tubes (see Materials and methods ) (b). The curve (c) corresponds to EVs remaining in the 12,000×g supernatant, which relates to sample (b) EVs. The 12,000×g pellets (a and b) are concentrated in PBS by factors of 20 and 100, respectively, whereas the 12,000×g supernatant (c) is as obtained after centrifugation.

Article Snippet: EVs, present in the 12,000×g pellet, are resuspended in a phosphate-buffered solution (PBS) [pH 7.4 without calcium and magnesium (Gibco)], with a 20 to 100 volume concentration factor (X), relative to the volume of the 2,000×g supernatant.

Techniques: Concentration Assay, Centrifugation

Journal: EBioMedicine

Article Title: Prognostic Impact of Circulating Tumor Cell Detected Using a Novel Fluidic Cell Microarray Chip System in Patients with Breast Cancer

doi: 10.1016/j.ebiom.2016.07.027

Figure Lengend Snippet: Workflow of sample processing and movement of cells in the FCMC device. (A) Workflow of sample processing and CTC detection. (a) Blood sample collection and enrichment using Ficoll-Paque PLUS. (b) The cell suspension is loaded into the FCMC device. The cells form monolayers in the microchambers and are immunostained by using a syringe pump. (c) All microchambers are scanned using a fluorescent microscope. (d) CTC are detected from the fluorescent images. (B) Movement of cells in the FCMC device. (a) The new CM chip, which is made from polystyrene (PS), is cleaned with UV ozone and coated with BSA. (b) The FCMC device is washed with PBS. (c) A cell suspension (68 uL) that is equivalent to 0.4 mL of blood is loaded into one FCMC device. (d) After “Suction for Trapping”, untrapped cells move into downstream microchambers and settle down on the bottom of these microchambers. The blue arrows indicate flow. (e) After “Suction for Monolayer”, overlapped cells are discharged from microchambers and move to the outside of microchambers or to the inside of downstream microchambers. (f) After repetitions of (d) and (e), all cells are trapped in a monolayer. (g), (h), (i) Immunostaining solutions are loaded, incubated and washed.

Article Snippet: The second immunostaining solution included 1:500 diluted Alexa Fluor 488 Goat Anti-Mouse IgG1 (γ1) (Life Technologies, Grand Island, NY), 1:500 diluted Alexa Fluor 647 Goat Anti-Mouse IgG2a (γ2a) (Life technologies) and 1:1000 diluted Hoechst 33342 10 mg/mL solution (Life technologies) in PBS solution containing 1% Tween 20 and 3% BSA.

Techniques: Microscopy, Chromatin Immunoprecipitation, Flow Cytometry, Immunostaining, Incubation