Journal: Science Advances

Article Title: Life span extension by glucose restriction is abrogated by methionine supplementation: Cross-talk between glucose and methionine and implication of methionine as a key regulator of life span

doi: 10.1126/sciadv.aba1306

Figure Lengend Snippet: ( A ) Fold changes of mRNA and translational efficiency for amino acid biosynthetic enzymes and transporters in two independent experiments using different GR protocols (see Materials and Methods). ( B ) Change of the protein expression of the major methionine transporter Mup1, measured by following the Mup1-GFP reporter in single cells in the microfluidic device. Shown is the average over 23 cells; each was normalized by the mean fluorescence before the switch. The arrow indicates the time point at which the media were switched from SD to GR. ( C to F ) Change of the intracellular concentration of amino acids after switching from SD to GR. (C and D) Ratios of intracellular amino acid concentrations between GR and SD in the BY4741 strain and BY4742 strain, respectively. Different colors represent different independent experiments. Box plot shows maximum, minimum, median, and top and bottom 20 percentile. (E and F) Amino acid concentration plotted against the usage in the proteome (genome frequency weighted by protein abundance). The red line is a linear fit excluding five charged amino acids (Arg, Lys, His, Glu, and Asp) and one polar amino acid (Gln). There is a log-linear correlation between the intracellular concentration and the usage.

Article Snippet: For life span measurement using the microfluidic device, images were taken once every 15 min by a Nikon TE2000 microscope or a Zeiss Axio Observer Z1 with 40× objective.

Techniques: Expressing, Fluorescence, Concentration Assay, Quantitative Proteomics

Journal: eLife

Article Title: Single cell analysis reveals immune cell–adipocyte crosstalk regulating the transcription of thermogenic adipocytes

doi: 10.7554/eLife.49501

Figure Lengend Snippet: ( A ) Workflow showing DAPI-stained nuclei pre- and post-FACS/filtration that underwent microfluidic partitioning and library preparation in the 10X genomics platform followed by sequencing using an Illumina HiSeq 4000. ( B ) tSNE-plot showing 14 clusters from ~6000 adipocytes derived from iWAT of mice exposed to cold for 24 hr. Each colored dot is an adipocyte assigned to a cluster based on transcriptomic signature. ( C ) Normalized expression values of the top two adipocyte subtype-specific cluster genes from ( B ) plotted as violin plots with clusters as rows and genes as columns. ( D ) tSNE-plot showing cluster-specific expression of selected marker genes from ( C ). ( E ) Normalized expression values of indicated genes in subtype-specific clusters plotted as violin plots with clusters as rows and genes as columns. Black arrow is pointing toward metabolically active Type nine adipocyte cluster and enriched gene.

Article Snippet: Commercial kit , FlowJEM aquapel-treated microfluidic device , FlowJEM , N/A , .

Techniques: Staining, Filtration, Sequencing, Derivative Assay, Expressing, Marker, Metabolic Labelling

Journal: eLife

Article Title: Single cell analysis reveals immune cell–adipocyte crosstalk regulating the transcription of thermogenic adipocytes

doi: 10.7554/eLife.49501

Figure Lengend Snippet:

Article Snippet: Commercial kit , FlowJEM aquapel-treated microfluidic device , FlowJEM , N/A , .

Techniques: Isolation, Multiplex Assay, Lysis, Software

Journal: bioRxiv

Article Title: Neurons as biosensors for discriminating neurological disorders in a brain-on-chip platform: Application to Alzheimer’s Disease using patient CSF

doi: 10.1101/2024.08.23.609425

Figure Lengend Snippet: Microfluidic platform by NETRI with Axion Biosystem’s technology. (a) Pictures of the NeoBento Pharma composed of four QuarterBento and sixteen microfluidic chips. (b) Rendering of zoomed QuarterBento of four DuaLink chip MEA. (c) Realistic rendering of a detailed DuaLink chip MEA showing three culture compartments. Black dots represent microelectrodes placed below the chip. (d) Schematic representation of DuaLink MEA with compartmentalized neuronal culture with one cell type seeded in Ch1 and the other cell type seeded in Ch3, axonal propagations are respectively in microchannels 1µ2 and 2µ3.

Article Snippet: Moreover, microfluidic devices were directly bonded on a MEA sheet developed by Axion Biosystems allowing neurons to be in direct contact with electrodes. ( ).

Techniques:

Journal: bioRxiv

Article Title: Neurons as biosensors for discriminating neurological disorders in a brain-on-chip platform: Application to Alzheimer’s Disease using patient CSF

doi: 10.1101/2024.08.23.609425

Figure Lengend Snippet: Characterization of co-culture of human Glutamatergic and GABAergic neurons in microfluidic device. (a) Timeline of cell culture maintenance with milestones during culture. (b) Brightfield contrast pictures of human glutamatergic and GABAergic neurons seeded in Dualink MEA at day 16. (c) Fluorescent picture of Live/Dead assay in the co-culture. Live cells were stained in green, and dead cells in red. (d, e) Immunofluorescent pictures of respectively, pluripotency markers (d), and specific neuronal markers (e) for each cell type. Sox2 was marked in red and Nestin in green. Β-III-tubulin was marked in green, vGlut1 in yellow, and GABA in red. In each immunofluorescent picture, DAPI was in blue. (f, g, h) Graph bars showing the quantification in channel 1 and channel 3 of the Live/dead assay, pluripotency markers and neuronal specific markers, respectively. (i) Raster plots from electrophysiological recordings at day 7, day 14 and day 21, showing spikes in time per electrode, in each DuaLink compartment. (j, k) Bar graphs showing, respectively weighted mean firing rate (WFR) and Synchrony index at day 7, day 14, and day 21 measured in channels and microchannels. * p-value< 0.05, *** p-value < 0.001 and **** p-value < 0.001. Error bar = SD.

Article Snippet: Moreover, microfluidic devices were directly bonded on a MEA sheet developed by Axion Biosystems allowing neurons to be in direct contact with electrodes. ( ).

Techniques: Co-Culture Assay, Cell Culture, Live Dead Assay, Staining