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Supplementary Figure S1. High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures
Juliette Ohan, Benjamin Pellea, , , Blake Hovde, , Armand E.K.Dichosa & Shawn R. Starkenburg
Pre-cultivation targeted FACS enrichment of
aGMDs and abGMDs. (A) Only aGMDs and abGMDs containing algal/bacterial cells
were first sorted by size by measuring forward scatter (FSC, x-axis at linear
scale) versus side scatter (SSC, y-axis at logarithmic scale). (B) aGMDs and
abGMDs were further analyzed by chlorophyll autofluorescence using a 650/50
nm filter (x-axis at logarithmic scale) versus 530/40 nm filter (y-axis at logarithmic
scale), to select for GMDs containing algal/bacterial cells. (C) aGMDs and...
Supplementary Figure S3. High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures
Juliette Ohan, Benjamin Pellea, , , Blake Hovde, , Armand E.K.Dichosa & Shawn R. Starkenburg
Post-cultivation
targeted FACS enrichment of abGMDs. Control population of cultivated aGMDs (A)
and experimental cultivated population of abGMDs (D) using FSC (x-axis; linear
scale) versus SSC (y-axis; log scale). (E) Cultivated abGMDs exhibiting chlorophyll
autofluorescence higher than that of the cultivated control aGMDs shown in (B)
using a 650/50 nm filter (x-axis at logarithmic scale) versus 530/40 nm filter
(y-axis at logarithmic scale). (F) Gated selection of cultivated abGMDs
exhibiting higher FSC (to distinguish from...
Supplementary Figure S2. High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures
Juliette Ohan, Benjamin Pellea, , , Blake Hovde, , Armand E.K.Dichosa & Shawn R. Starkenburg
Schematic for cultivation chamber. The
cultivation chamber was fabricated and sterilized as described for the
microfluidic droplet generator chip (Figure 2 legend). The microfluidic
cultivation chamber was used to microscopically visualize droplets without
disturbing them or sacrificing viability, as well as providing an even
distribution of light.
Supplementary figure 2. A mismatch-tolerant RT-quantitative PCR: application to broad-spectrum detection of respiratory syncytial virus
, , , Yi Zhou, Qin Chen &
Maximum likelihood (ML) tree of sequences from
four clinical samples. ML tree of RSV-A and RSV-B sequences obtained from four
clinical samples (Sp16, Sp19, Sp8 and Sp9) in this study. The reference
sequences were retrieved from GenBank.
Supplementary figure 1. Fluidic resistance control enables high throughput establishment of mixed species biofilms
Mads Frederik Hansen, Anders Meyer Torp, Jonas Stenløkke Madsen, Henriette Lyng Røder & Mette Burmølle
Viability is
unaffected by short time incubation in high viscosity medium. (A) The number of
counted CFU/ml of Pseudomonas putida and Paenibacillus amylolyticus is
identical after 45 min. of incubation in PBS and high viscosity liquid (50%
glycerol). (B) Similarly, the viability of Xanthomonas retroflexus was
unaffected by incubation in high viscosity liquid and did not change over a
short time period (N=3, Error bars represent Standard Error (SE)).
Supplementary Table S1. High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures
Juliette Ohan, Benjamin Pellea, , , Blake Hovde, , Armand E.K.Dichosa & Shawn R. Starkenburg
Cell input
modifications for GMD capture. Bacterial and algal concentrations were
adjusted to cell concentrations OD600, OD750 in 1x PBS and 100 µl of the
resultant cell suspension (or 50 + 50 µL for abGMDs) was used as cell input
for initial capture in the CelGel Matrix prior to sorting. This ratio of
bacteria-to-matrix or algae-to-matrix (# cells/GMD) was verified by
microscopy; data not presented. “aGMD” = algae only GMDs, “bGMD” = bacteria only
GMDs,...
Supplementary Figure S1. High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures
Juliette Ohan, Benjamin Pellea, , , Blake Hovde, , Armand E.K.Dichosa & Shawn R. Starkenburg
Pre-cultivation targeted FACS enrichment of
aGMDs and abGMDs. (A) Only aGMDs and abGMDs containing algal/bacterial cells
were first sorted by size by measuring forward scatter (FSC, x-axis at linear
scale) versus side scatter (SSC, y-axis at logarithmic scale). (B) aGMDs and
abGMDs were further analyzed by chlorophyll autofluorescence using a 650/50
nm filter (x-axis at logarithmic scale) versus 530/40 nm filter (y-axis at logarithmic
scale), to select for GMDs containing algal/bacterial cells. (C) aGMDs and...
Supplementary figure 1. A mismatch-tolerant RT-quantitative PCR: application to broad-spectrum detection of respiratory syncytial virus
, , , Yi Zhou, Qin Chen &
Comparison of the new
mismatch-tolerant RT-qPCR with the assays with only Taq (conventional) and high
fidelity (Q5) DNA polymerase. A. Detection of RSV mutants; B. Detection HCoV
229E mutants. The amplified products were detected using 2.5% agarose gel electrophoresis.
The arrow highlights the specific product. For other details, please see Figure
3.
Supplementary Table S1. High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures
Juliette Ohan, Benjamin Pellea, , , Blake Hovde, , Armand E.K.Dichosa & Shawn R. Starkenburg
Cell input
modifications for GMD capture. Bacterial and algal concentrations were
adjusted to cell concentrations OD600, OD750 in 1x PBS and 100 µl of the
resultant cell suspension (or 50 + 50 µL for abGMDs) was used as cell input
for initial capture in the CelGel Matrix prior to sorting. This ratio of
bacteria-to-matrix or algae-to-matrix (# cells/GMD) was verified by
microscopy; data not presented. “aGMD” = algae only GMDs, “bGMD” = bacteria only
GMDs,...
Supplementary Figure S2. High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures
Juliette Ohan, Benjamin Pellea, , , Blake Hovde, , Armand E.K.Dichosa & Shawn R. Starkenburg
Schematic for cultivation chamber. The
cultivation chamber was fabricated and sterilized as described for the
microfluidic droplet generator chip (Figure 2 legend). The microfluidic
cultivation chamber was used to microscopically visualize droplets without
disturbing them or sacrificing viability, as well as providing an even
distribution of light.
Supplementary figure 2. A mismatch-tolerant RT-quantitative PCR: application to broad-spectrum detection of respiratory syncytial virus
, , , Yi Zhou, Qin Chen &
Maximum likelihood (ML) tree of sequences from
four clinical samples. ML tree of RSV-A and RSV-B sequences obtained from four
clinical samples (Sp16, Sp19, Sp8 and Sp9) in this study. The reference
sequences were retrieved from GenBank.
Supplementary Figure S3. High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures
Juliette Ohan, Benjamin Pellea, , , Blake Hovde, , Armand E.K.Dichosa & Shawn R. Starkenburg
Post-cultivation
targeted FACS enrichment of abGMDs. Control population of cultivated aGMDs (A)
and experimental cultivated population of abGMDs (D) using FSC (x-axis; linear
scale) versus SSC (y-axis; log scale). (E) Cultivated abGMDs exhibiting chlorophyll
autofluorescence higher than that of the cultivated control aGMDs shown in (B)
using a 650/50 nm filter (x-axis at logarithmic scale) versus 530/40 nm filter
(y-axis at logarithmic scale). (F) Gated selection of cultivated abGMDs
exhibiting higher FSC (to distinguish from...
Supplementary figure 1. A mismatch-tolerant RT-quantitative PCR: application to broad-spectrum detection of respiratory syncytial virus
, , , Yi Zhou, Qin Chen &
Comparison of the new
mismatch-tolerant RT-qPCR with the assays with only Taq (conventional) and high
fidelity (Q5) DNA polymerase. A. Detection of RSV mutants; B. Detection HCoV
229E mutants. The amplified products were detected using 2.5% agarose gel electrophoresis.
The arrow highlights the specific product. For other details, please see Figure
3.
Supplementary figure 1. Fluidic resistance control enables high throughput establishment of mixed species biofilms
Mads Frederik Hansen, Anders Meyer Torp, Jonas Stenløkke Madsen, Henriette Lyng Røder & Mette Burmølle
Viability is
unaffected by short time incubation in high viscosity medium. (A) The number of
counted CFU/ml of Pseudomonas putida and Paenibacillus amylolyticus is
identical after 45 min. of incubation in PBS and high viscosity liquid (50%
glycerol). (B) Similarly, the viability of Xanthomonas retroflexus was
unaffected by incubation in high viscosity liquid and did not change over a
short time period (N=3, Error bars represent Standard Error (SE)).
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