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u2os cells human bone osteosarcoma cell line (DSMZ)

Name: u2os cells human bone osteosarcoma cell line
Brand: DSMZ
Rating: 95 (116 reviews)

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DSMZ u2os cells human bone osteosarcoma cell line
U2os Cells Human Bone Osteosarcoma Cell Line, supplied by DSMZ, used in various techniques. Bioz Stars score: 95/100, based on 116 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/u2os cells human bone osteosarcoma cell line/product/DSMZ
Average 95 stars, based on 116 article reviews

u2os cells human bone osteosarcoma cell line - by Bioz Stars, 2026-03

95/100 stars

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(A) Schematic representation of the time-of-drug-addition assay. U2OS cells were treated with (B) 2 µM JG98 or (C) 1 µM JG345 at the indicated time points (conditions 1-8) or with the DMSO control. Virus inoculum (CHIKV-LR at MOI 1) was present for 1.5 h, after which the inoculum was removed, and fresh medium was added with or without the Hsp70 inhibitors or DMSO control. At 9 hpi, the infectious virus particle production was assessed using the plaque assay. Data are presented as mean±SEM from at least three independent experiments. One-way ANOVA was used to evaluate statistical differences from the non-treated control and was presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Journal: bioRxiv

Article Title: The Heat shock protein 70 machinery is crucial in the production of infectious chikungunya virus progeny

doi: 10.64898/2026.01.26.701664

Figure Lengend Snippet: (A) Schematic representation of the time-of-drug-addition assay. U2OS cells were treated with (B) 2 µM JG98 or (C) 1 µM JG345 at the indicated time points (conditions 1-8) or with the DMSO control. Virus inoculum (CHIKV-LR at MOI 1) was present for 1.5 h, after which the inoculum was removed, and fresh medium was added with or without the Hsp70 inhibitors or DMSO control. At 9 hpi, the infectious virus particle production was assessed using the plaque assay. Data are presented as mean±SEM from at least three independent experiments. One-way ANOVA was used to evaluate statistical differences from the non-treated control and was presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Article Snippet: The human bone osteosarcoma epithelial cell line U2OS (ATCC HTB-96) and human foreskin fibroblast cell line HFF-1 (ATCC SCRC-1041) were maintained in Dulbecco’s minimal essential medium (DMEM), high glucose, GlutaMAX tm, and sodium pyruvate (Gibco), supplemented with penicillin-streptomycin (P/S; 100U/mL, Gibco) and 10% or 15% fetal bovine serum (FBS, Lonza), respectively.

Techniques: Control, Virus, Plaque Assay

(A) U2OS cells were treated with increasing concentrations of the Hsp70 inhibitors or the equivalent volume of the DMSO control. The metabolic activity of the cells was assessed after 16 hours (h) of treatment using the MTS assay and was normalized to the DMSO control. A decrease of 15% in metabolically active cells was considered non-toxic (dotted line represents 85%). (B-F) U2OS cells were infected with CHIKV-LR OPY1 at multiplicity of infection (MOI) 1 in the presence of increasing concentrations of the (B) DMSO control (0.002% matches 0.1 µM JG-compound, 0.01% DMSO matches 0.5 µM JG-compound, etc.) or the Hsp70 inhibitors concentrations of (C) JG18, (D) JG40, (E) JG98, (F) JG345. Supernatants were collected at 9 hpi, and the number of infectious CHIKV particles was quantified using plaque assay on Vero-WHO cells. (B-F) Percentage inhibition was determined relative to the DMSO control, and IC50 (which corresponds to a 50% reduction in viral titer) per Hsp70 inhibitor was determined via non-linear regression. Data are presented as mean±SEM from three independent experiments. (B) Statistical differences were determined using One-way ANOVA and were presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Journal: bioRxiv

Article Title: The Heat shock protein 70 machinery is crucial in the production of infectious chikungunya virus progeny

doi: 10.64898/2026.01.26.701664

Figure Lengend Snippet: (A) U2OS cells were treated with increasing concentrations of the Hsp70 inhibitors or the equivalent volume of the DMSO control. The metabolic activity of the cells was assessed after 16 hours (h) of treatment using the MTS assay and was normalized to the DMSO control. A decrease of 15% in metabolically active cells was considered non-toxic (dotted line represents 85%). (B-F) U2OS cells were infected with CHIKV-LR OPY1 at multiplicity of infection (MOI) 1 in the presence of increasing concentrations of the (B) DMSO control (0.002% matches 0.1 µM JG-compound, 0.01% DMSO matches 0.5 µM JG-compound, etc.) or the Hsp70 inhibitors concentrations of (C) JG18, (D) JG40, (E) JG98, (F) JG345. Supernatants were collected at 9 hpi, and the number of infectious CHIKV particles was quantified using plaque assay on Vero-WHO cells. (B-F) Percentage inhibition was determined relative to the DMSO control, and IC50 (which corresponds to a 50% reduction in viral titer) per Hsp70 inhibitor was determined via non-linear regression. Data are presented as mean±SEM from three independent experiments. (B) Statistical differences were determined using One-way ANOVA and were presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Techniques: Control, Activity Assay, MTS Assay, Metabolic Labelling, Infection, Plaque Assay, Inhibition

U2OS cells were infected with the (A) African S27 CHIKV strain or the (B) Caribbean 99659 strain at MOI 1 and simultaneously treated with 10 µM, 10 µM, 2 µM, or 1 µM of JG18, JG40, JG98, or JG345, respectively, or the DMSO control. Production of infectious virus particles was assessed at 9 hpi. (C) U2OS cells were infected with DENV A2 (16681) at MOI 0.5 and treated with 10 µM JG18 or JG40, or the vehicle control. 24 hpi, supernatants were collected and DENV progeny production was analyzed via plaque assay on BHK-21 cells. Data are presented as mean±SEM from three independent experiments, and statistical differences were determined via one-way ANOVA and presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Journal: bioRxiv

Article Title: The Heat shock protein 70 machinery is crucial in the production of infectious chikungunya virus progeny

doi: 10.64898/2026.01.26.701664

Figure Lengend Snippet: U2OS cells were infected with the (A) African S27 CHIKV strain or the (B) Caribbean 99659 strain at MOI 1 and simultaneously treated with 10 µM, 10 µM, 2 µM, or 1 µM of JG18, JG40, JG98, or JG345, respectively, or the DMSO control. Production of infectious virus particles was assessed at 9 hpi. (C) U2OS cells were infected with DENV A2 (16681) at MOI 0.5 and treated with 10 µM JG18 or JG40, or the vehicle control. 24 hpi, supernatants were collected and DENV progeny production was analyzed via plaque assay on BHK-21 cells. Data are presented as mean±SEM from three independent experiments, and statistical differences were determined via one-way ANOVA and presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Techniques: Infection, Control, Virus, Plaque Assay

U2OS cells were treated with 2 µM JG98, 1 µM JG345, or the DMSO control during CHIKV infection at MOI 10. Virus production was measured at 9 hpi using a plaque assay. Data are presented as mean±SEM from at least three independent experiments. One-way ANOVA was used to evaluate statistical differences from the DMSO control and was presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Journal: bioRxiv

Article Title: The Heat shock protein 70 machinery is crucial in the production of infectious chikungunya virus progeny

doi: 10.64898/2026.01.26.701664

Figure Lengend Snippet: U2OS cells were treated with 2 µM JG98, 1 µM JG345, or the DMSO control during CHIKV infection at MOI 10. Virus production was measured at 9 hpi using a plaque assay. Data are presented as mean±SEM from at least three independent experiments. One-way ANOVA was used to evaluate statistical differences from the DMSO control and was presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Techniques: Control, Infection, Virus, Plaque Assay

(A) The total intracellular vRNA and (B) genomic vRNA copy number were assessed at 4, 6, and 8 hpi in U2OS cells infected with CHIKV at MOI 10 and treated with vehicle control DMSO or 2μM JG98. Intracellular vRNA copies were quantified by RT-qPCR using specific primers against (A) E1 and (B) nsP1. (C) The number of subgenomic RNA copies was determined by subtracting the genomic vRNA copies from the total vRNA copies. Data is presented as mean ± SEM from at least three independent experiments. Student T-test was used to evaluate statistical differences from the DMSO control per timepoint and were presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Journal: bioRxiv

Article Title: The Heat shock protein 70 machinery is crucial in the production of infectious chikungunya virus progeny

doi: 10.64898/2026.01.26.701664

Figure Lengend Snippet: (A) The total intracellular vRNA and (B) genomic vRNA copy number were assessed at 4, 6, and 8 hpi in U2OS cells infected with CHIKV at MOI 10 and treated with vehicle control DMSO or 2μM JG98. Intracellular vRNA copies were quantified by RT-qPCR using specific primers against (A) E1 and (B) nsP1. (C) The number of subgenomic RNA copies was determined by subtracting the genomic vRNA copies from the total vRNA copies. Data is presented as mean ± SEM from at least three independent experiments. Student T-test was used to evaluate statistical differences from the DMSO control per timepoint and were presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Techniques: Infection, Control, Quantitative RT-PCR

Gating strategy to determine the percentage of cells positive for CHIKV E2 and capsid expression in U2OS cells treated with JG98, JG345, or the DMSO control. (A) Gating for cells and exclusion of doublets. (B and C) Gating to determine the percentage of cells positive for (B) capsid and (C) E2 based on mock-infected cells.

Journal: bioRxiv

Article Title: The Heat shock protein 70 machinery is crucial in the production of infectious chikungunya virus progeny

doi: 10.64898/2026.01.26.701664

Figure Lengend Snippet: Gating strategy to determine the percentage of cells positive for CHIKV E2 and capsid expression in U2OS cells treated with JG98, JG345, or the DMSO control. (A) Gating for cells and exclusion of doublets. (B and C) Gating to determine the percentage of cells positive for (B) capsid and (C) E2 based on mock-infected cells.

Techniques: Expressing, Control, Infection

(A-D) U2OS cells were infected with CHIKV at MOI 10 and treated with JG98 (2μM), JG345 (1μM), or the DMSO control. At 9hpi, E2 and capsid expression were assessed using flow cytometry to determine the (A and C) percentage of positive cells and (B and D) mean fluorescent intensity (MFI; geometric mean). (E and F) Representative western blot of capsid, E2, or vinculin expression from protein lysates of U2OS cells infected with CHIKV at MOI 10 and treated for 9 h with Hsp70 inhibitor JG-98 (2μM), JG-345 (1μM), or the DMSO control. (G and H) Quantification of Western blots from three independent experiments. Protein levels are normalized to vinculin and are expressed as relative protein level to vehicle control DMSO. Data are presented as mean±SEM from at least three independent experiments. One-way ANOVA was used to evaluate statistical differences from the DMSO control and was presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Journal: bioRxiv

Article Title: The Heat shock protein 70 machinery is crucial in the production of infectious chikungunya virus progeny

doi: 10.64898/2026.01.26.701664

Figure Lengend Snippet: (A-D) U2OS cells were infected with CHIKV at MOI 10 and treated with JG98 (2μM), JG345 (1μM), or the DMSO control. At 9hpi, E2 and capsid expression were assessed using flow cytometry to determine the (A and C) percentage of positive cells and (B and D) mean fluorescent intensity (MFI; geometric mean). (E and F) Representative western blot of capsid, E2, or vinculin expression from protein lysates of U2OS cells infected with CHIKV at MOI 10 and treated for 9 h with Hsp70 inhibitor JG-98 (2μM), JG-345 (1μM), or the DMSO control. (G and H) Quantification of Western blots from three independent experiments. Protein levels are normalized to vinculin and are expressed as relative protein level to vehicle control DMSO. Data are presented as mean±SEM from at least three independent experiments. One-way ANOVA was used to evaluate statistical differences from the DMSO control and was presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Techniques: Infection, Control, Expressing, Flow Cytometry, Western Blot

U2OS cells were infected with the (A-C, G-I) CHIKV-LR or the (D-F) CHIKV S27 strain for 9 h in the presence of (A-F) 2 µM JG98, 1 µM JG345, (G-I) 20 µM VER-155008, or the DMSO control. Supernatants were collected and the number of (A, D, and G) infectious particles and (B, E, and H) secreted genome equivalent copies (GECs) were measured using plaque assay and RT-qPCR, respectively. (C, F, and I) Specific infectivity is depicted as the ratio between produced infectious particles and GECs. Data are presented as mean± SEM from three independent experiments. One-way ANOVA was used to evaluate statistical differences from the DMSO control and was given when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Journal: bioRxiv

Article Title: The Heat shock protein 70 machinery is crucial in the production of infectious chikungunya virus progeny

doi: 10.64898/2026.01.26.701664

Figure Lengend Snippet: U2OS cells were infected with the (A-C, G-I) CHIKV-LR or the (D-F) CHIKV S27 strain for 9 h in the presence of (A-F) 2 µM JG98, 1 µM JG345, (G-I) 20 µM VER-155008, or the DMSO control. Supernatants were collected and the number of (A, D, and G) infectious particles and (B, E, and H) secreted genome equivalent copies (GECs) were measured using plaque assay and RT-qPCR, respectively. (C, F, and I) Specific infectivity is depicted as the ratio between produced infectious particles and GECs. Data are presented as mean± SEM from three independent experiments. One-way ANOVA was used to evaluate statistical differences from the DMSO control and was given when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Techniques: Infection, Control, Plaque Assay, Quantitative RT-PCR, Produced

Metabolic activity of the U2OS cells was assessed after 16 hours of treatment with increasing concentrations of the VER155008 or the equivalent volume of the DMSO control. The percentage of metabolically active cells relative to the DMSO control. A decrease of 15% in metabolically active cells was considered non-toxic (dotted line represents 85%). Data are presented as mean±SEM from three independent experiments.

Journal: bioRxiv

Article Title: The Heat shock protein 70 machinery is crucial in the production of infectious chikungunya virus progeny

doi: 10.64898/2026.01.26.701664

Figure Lengend Snippet: Metabolic activity of the U2OS cells was assessed after 16 hours of treatment with increasing concentrations of the VER155008 or the equivalent volume of the DMSO control. The percentage of metabolically active cells relative to the DMSO control. A decrease of 15% in metabolically active cells was considered non-toxic (dotted line represents 85%). Data are presented as mean±SEM from three independent experiments.

Techniques: Activity Assay, Control, Metabolic Labelling

Metabolic activity of (A) U2OS and (B) HFF-1 cells was assessed after 16 h of treatment with increasing concentrations of JG231 or the equivalent volume of the DMSO control. The percentage of metabolically active cells is presented relative to the DMSO control. A decrease of 15% in metabolically active cells was considered non-toxic (dotted line represents 85%). (C) U2OS cells and (D) HFF-1 cells were infected with CHIKV-LR for 9 h in the presence of 2 µM JG231 or the DMSO control. Supernatants were collected, and the number of infectious particles in the supernatant was determined using the plaque assay. (E and F) Mouse skin explants were infected with 10 6 PFU/mL of the CHIKV 899 strain during treatment with 2 µM JG231 or an equal volume of DMSO. (E) Infectious virus production (tissue culture infectious dose 50% (TCID 50 )) and (F) total virus production (GEC) at 2 dpi is displayed per mg tissue. The dotted line indicates the limit of quantification (LOQ). To evaluate statistical differences from the DMSO control, we used a Student T-test, and differences are presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Journal: bioRxiv

Article Title: The Heat shock protein 70 machinery is crucial in the production of infectious chikungunya virus progeny

doi: 10.64898/2026.01.26.701664

Figure Lengend Snippet: Metabolic activity of (A) U2OS and (B) HFF-1 cells was assessed after 16 h of treatment with increasing concentrations of JG231 or the equivalent volume of the DMSO control. The percentage of metabolically active cells is presented relative to the DMSO control. A decrease of 15% in metabolically active cells was considered non-toxic (dotted line represents 85%). (C) U2OS cells and (D) HFF-1 cells were infected with CHIKV-LR for 9 h in the presence of 2 µM JG231 or the DMSO control. Supernatants were collected, and the number of infectious particles in the supernatant was determined using the plaque assay. (E and F) Mouse skin explants were infected with 10 6 PFU/mL of the CHIKV 899 strain during treatment with 2 µM JG231 or an equal volume of DMSO. (E) Infectious virus production (tissue culture infectious dose 50% (TCID 50 )) and (F) total virus production (GEC) at 2 dpi is displayed per mg tissue. The dotted line indicates the limit of quantification (LOQ). To evaluate statistical differences from the DMSO control, we used a Student T-test, and differences are presented when p≤0.05 with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.001.

Techniques: Activity Assay, Control, Metabolic Labelling, Infection, Plaque Assay, Virus

IPI measurement of LDs in U2OS cells. (A) Single-color (left) and two-color (right) IP images at 2193 cm −1 and 2855 cm −1 . The two-color IP image was constructed by superimposing IP images simultaneously obtained with IR excitations of 2855 cm −1 (45 kHz) and 2193 cm −1 (50 kHz), which are marked by false green and red colors, respectively. The IPI contrasts were adjusted to reveal the cellular structures. The inset image is the bright-field image of the corresponding region. Cutoff frequency, 70 Hz. Step size, 200 nm. Pixel dwell time, 2 ms. Probe power, 15 mW. IR excitation power, 0.2 mW (at 2193 cm −1 ) and 0.1 mW (at 2855 cm −1 ). Scale bars, 20 μm. (B) IP spectra of LDs before and after background calibration. Three LDs are indicated in . Each data point in the IP spectra was obtained by 50 ms averaging and normalization to the intensity of the IR excitation pulse. The spectra on the bottom were calibrated by matching the background signal level in the off-resonant spectral region (2740 cm −1 to 2760 cm −1 ). Stepsize, 1 cm −1 . (C) Histogram for IP intensity at 2855 cm −1 of LDs. We obtained IP spectra of 47 LDs within U2OS cells from seven different regions, and the histogram shows the distribution of IP intensities at 2855 cm −1 of the calibrated IP spectra.

Journal: Chemical Science

Article Title: Monitoring the synthesis of neutral lipids in lipid droplets of living human cancer cells using two-color infrared photothermal microscopy †

doi: 10.1039/d3sc04705a

Figure Lengend Snippet: IPI measurement of LDs in U2OS cells. (A) Single-color (left) and two-color (right) IP images at 2193 cm −1 and 2855 cm −1 . The two-color IP image was constructed by superimposing IP images simultaneously obtained with IR excitations of 2855 cm −1 (45 kHz) and 2193 cm −1 (50 kHz), which are marked by false green and red colors, respectively. The IPI contrasts were adjusted to reveal the cellular structures. The inset image is the bright-field image of the corresponding region. Cutoff frequency, 70 Hz. Step size, 200 nm. Pixel dwell time, 2 ms. Probe power, 15 mW. IR excitation power, 0.2 mW (at 2193 cm −1 ) and 0.1 mW (at 2855 cm −1 ). Scale bars, 20 μm. (B) IP spectra of LDs before and after background calibration. Three LDs are indicated in . Each data point in the IP spectra was obtained by 50 ms averaging and normalization to the intensity of the IR excitation pulse. The spectra on the bottom were calibrated by matching the background signal level in the off-resonant spectral region (2740 cm −1 to 2760 cm −1 ). Stepsize, 1 cm −1 . (C) Histogram for IP intensity at 2855 cm −1 of LDs. We obtained IP spectra of 47 LDs within U2OS cells from seven different regions, and the histogram shows the distribution of IP intensities at 2855 cm −1 of the calibrated IP spectra.

Article Snippet: U2OS cell line (human bone osteosarcoma epithelial cells) and Huh-7 cell line (human hepatocellular carcinoma cells) were supplied from Korean Cell Line Bank (Seoul, Korea) and cultured in DMEM (Welgene, LM 001-17) supplemented with penicillin (100 U mL −1 ), streptomycin (100 μg mL −1 ), 10% heat-inactivated fetal bovine serum (Welgene, S1010-01) at 37 °C and pH 7.4 (5% CO 2 ).

Techniques: Construct

Two-color IPI measurements of U2OS cells cultured exposed to PA and PA-d 31 . (A) Two-color IP images of fixed U2OS cells cultured in different growth media (250 μM PA, 250 μM PA-d 31 , and standard medium) for 24 hours. The green and red false colors indicate IPI contrasts of 2855 cm −1 and 2193 cm −1 , respectively. Cutoff frequency, 70 Hz. Step size, 200 nm. Pixel dwell time, 2 ms. Probe power, 15 mW. IR excitation power, 0.2 mW (at 2193 cm −1 ) and 0.1 mW (at 2855 cm −1 ). Scale bar, 20 μm. (B) Representative IP spectra of LDs observed in at two different spectral regions. Each data point in the IP spectra was obtained by 50 ms averaging and normalization to the intensity of the IR excitation pulse. Stepsize, 1 cm −1 .

Journal: Chemical Science

Article Title: Monitoring the synthesis of neutral lipids in lipid droplets of living human cancer cells using two-color infrared photothermal microscopy †

doi: 10.1039/d3sc04705a

Figure Lengend Snippet: Two-color IPI measurements of U2OS cells cultured exposed to PA and PA-d 31 . (A) Two-color IP images of fixed U2OS cells cultured in different growth media (250 μM PA, 250 μM PA-d 31 , and standard medium) for 24 hours. The green and red false colors indicate IPI contrasts of 2855 cm −1 and 2193 cm −1 , respectively. Cutoff frequency, 70 Hz. Step size, 200 nm. Pixel dwell time, 2 ms. Probe power, 15 mW. IR excitation power, 0.2 mW (at 2193 cm −1 ) and 0.1 mW (at 2855 cm −1 ). Scale bar, 20 μm. (B) Representative IP spectra of LDs observed in at two different spectral regions. Each data point in the IP spectra was obtained by 50 ms averaging and normalization to the intensity of the IR excitation pulse. Stepsize, 1 cm −1 .

Techniques: Cell Culture

$Investigation of the LDs with time. (A) Two-color (excitation: 2193 cm −1 and 2855 cm −1 ) IP images of fixed U2OS cells at different time points after PA-d 31 administration. PA-d 31 treatment times were indicated in the upper left corner of each image. The color scheme is the same as in . Cutoff frequency, 70 Hz. Step size, 200 nm. Pixel dwell time, 2 ms. Probe power, 15 mW. IR excitation power, 0.2 mW (at 2193 cm −1 ) and 0.1 mW (at 2855 cm −1 ). Scale bars, 20 μm. (B) IP spectra of LDs at time points (20 min, 1 h, 6 h, and 24 h) after PA-d 31 administration. Each data point in the IP spectra was obtained by 50 ms averaging and normalization to the intensity of the IR excitation pulse. Stepsize, 1 cm −1 . The y -axis scales in the two regions (2030 cm −1 to 2230 cm −1 and 2750 cm −1 to 2950 cm −1 ) are adjusted differently to enhance clarity. (C) Two-color IP images of fixed U2OS cells treated with PA-d 31 for 3 hours. The experimental condition is the same as that of . Scale bars, 20 μm. The figures below depict magnified images of five LDs with 2 μm scale bars. (D) Mole fractions of CD 2 and CH 2 groups from neutral lipids of . Two wavelengths of 2193 cm −1 and 2855 cm −1 were chosen to calculate the mole fraction. (E) Temporal change in mole fraction. The mole fraction of 10 different LDs was averaged at each time point. The error bars represent the upper and lower bounds of the mole fraction, corresponding to the maximum and minimum values, respectively.$

Journal: Chemical Science

Article Title: Monitoring the synthesis of neutral lipids in lipid droplets of living human cancer cells using two-color infrared photothermal microscopy †

doi: 10.1039/d3sc04705a

Figure Lengend Snippet: Investigation of the LDs with time. (A) Two-color (excitation: 2193 cm −1 and 2855 cm −1 ) IP images of fixed U2OS cells at different time points after PA-d 31 administration. PA-d 31 treatment times were indicated in the upper left corner of each image. The color scheme is the same as in . Cutoff frequency, 70 Hz. Step size, 200 nm. Pixel dwell time, 2 ms. Probe power, 15 mW. IR excitation power, 0.2 mW (at 2193 cm −1 ) and 0.1 mW (at 2855 cm −1 ). Scale bars, 20 μm. (B) IP spectra of LDs at time points (20 min, 1 h, 6 h, and 24 h) after PA-d 31 administration. Each data point in the IP spectra was obtained by 50 ms averaging and normalization to the intensity of the IR excitation pulse. Stepsize, 1 cm −1 . The y -axis scales in the two regions (2030 cm −1 to 2230 cm −1 and 2750 cm −1 to 2950 cm −1 ) are adjusted differently to enhance clarity. (C) Two-color IP images of fixed U2OS cells treated with PA-d 31 for 3 hours. The experimental condition is the same as that of . Scale bars, 20 μm. The figures below depict magnified images of five LDs with 2 μm scale bars. (D) Mole fractions of CD 2 and CH 2 groups from neutral lipids of . Two wavelengths of 2193 cm −1 and 2855 cm −1 were chosen to calculate the mole fraction. (E) Temporal change in mole fraction. The mole fraction of 10 different LDs was averaged at each time point. The error bars represent the upper and lower bounds of the mole fraction, corresponding to the maximum and minimum values, respectively.

Techniques:

Real-time monitoring of neutral lipid synthesis in living U2OS cells. (A) Time-lapse two-color (excitation: 2193 cm −1 and 2855 cm −1 ) IP images of a live U2OS cell. The two-color IP images were obtained every 10 minutes for 4 hours. The color scheme is the same as in . Cutoff frequency, 70 Hz. Step size, 400 nm. Pixel dwell time, 2 ms. Probe power, 15 mW. IR excitation power, 0.2 mW (at 2193 cm −1 ) and 0.1 mW (at 2855 cm −1 ). Scale bars, 20 μm. (B and C) Temporal IP intensity profiles of LDs in living U2OS cells. The maximal intensities of 2193 cm −1 and 2855 cm −1 of LD1 and 2 in are plotted with time, indicated as white dotted boxes. Linear lines fitted to the temporal profiles of each IP intensity are displayed as dotted lines to show their respective trends. (D) IP spectra of the two LDs. Each data point in the IP spectra was obtained by 50 ms averaging and normalization to the intensity of the IR excitation pulse. Step size, 1 cm −1 .

Journal: Chemical Science

Article Title: Monitoring the synthesis of neutral lipids in lipid droplets of living human cancer cells using two-color infrared photothermal microscopy †

doi: 10.1039/d3sc04705a

Figure Lengend Snippet: Real-time monitoring of neutral lipid synthesis in living U2OS cells. (A) Time-lapse two-color (excitation: 2193 cm −1 and 2855 cm −1 ) IP images of a live U2OS cell. The two-color IP images were obtained every 10 minutes for 4 hours. The color scheme is the same as in . Cutoff frequency, 70 Hz. Step size, 400 nm. Pixel dwell time, 2 ms. Probe power, 15 mW. IR excitation power, 0.2 mW (at 2193 cm −1 ) and 0.1 mW (at 2855 cm −1 ). Scale bars, 20 μm. (B and C) Temporal IP intensity profiles of LDs in living U2OS cells. The maximal intensities of 2193 cm −1 and 2855 cm −1 of LD1 and 2 in are plotted with time, indicated as white dotted boxes. Linear lines fitted to the temporal profiles of each IP intensity are displayed as dotted lines to show their respective trends. (D) IP spectra of the two LDs. Each data point in the IP spectra was obtained by 50 ms averaging and normalization to the intensity of the IR excitation pulse. Step size, 1 cm −1 .

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