Journal: Biophysical Journal

Article Title: Impact of staining on cell acoustic properties

doi: 10.1016/j.bpj.2025.11.007

Figure Lengend Snippet: Determination of the acoustic impedance of calcein-stained and unstained MCF-7 cells. ( a ) and ( b ) present the percentages of cells exhibiting positive, zero, and negative contrast factors across different iodixanol concentrations. ( c ) and ( d ) display the cell percentages at the pressure node ( blue dots ) and antinodes ( red dots ), with data fitted to the cumulative distribution function of a normal distribution. The transition iodixanol concentration is identified at the intersection of the two fitting curves, where the proportion of cells with contrast factor near zero is maximized. The acoustic impedance was determined to be 1.580 ± 0.017 (0.009) MPa · s m −1 (mean ± standard deviation (fitting error of the mean)) for calcein-stained MCF-7 cells and 1.622 ± 0.030 (0.013) MPa · s m −1 for unstained MCF-7 cells.

Article Snippet: To examine the impact of staining on cell physical properties, four fluorescent dyes were used: a cell-permeable tracer calcein AM (Fisher Scientific International, Pittsburgh, PA) for cytoplasm staining; anti-EpCAM FITC (Becton, Dickinson and Company, Franklin Lakes, NJ), which binds to the cell membranes; E-cadherin CD324 PE (Miltenyi Biotec Norden, Lund, Sweden), also for cell membrane binding; and Hoechst 33342 (TargetMol Chemicals, Boston, MA), which binds to DNA in the cell nucleus.

Techniques: Staining, Concentration Assay, Standard Deviation

Journal: Biophysical Journal

Article Title: Impact of staining on cell acoustic properties

doi: 10.1016/j.bpj.2025.11.007

Figure Lengend Snippet: Measured acoustic impedance of stained and unstained ( a ) DU145, ( b ) MCF-7, ( c ) Jurkat, and ( d ) BV2 cells over 9 hours after exposure to PBS solution. The incubation time for calcein-stained cells was 20 min. Colors represent the acoustic impedance measured at different time points, whereas error bars indicate the fitting error of the mean obtained from fitting the data to the cumulative distribution function of a normal function. Brackets indicate endpoint contrasts (1–3 h to 7–9 h) tested with two-sided Wald z-tests on the two fitted estimates. Asterisks denote significance ( ∗ p < 0.05; ns = not significant). p -values reflect within-experiment uncertainty of the fits.

Article Snippet: To examine the impact of staining on cell physical properties, four fluorescent dyes were used: a cell-permeable tracer calcein AM (Fisher Scientific International, Pittsburgh, PA) for cytoplasm staining; anti-EpCAM FITC (Becton, Dickinson and Company, Franklin Lakes, NJ), which binds to the cell membranes; E-cadherin CD324 PE (Miltenyi Biotec Norden, Lund, Sweden), also for cell membrane binding; and Hoechst 33342 (TargetMol Chemicals, Boston, MA), which binds to DNA in the cell nucleus.

Techniques: Staining, Incubation

Journal: Biophysical Journal

Article Title: Impact of staining on cell acoustic properties

doi: 10.1016/j.bpj.2025.11.007

Figure Lengend Snippet: Measured acoustic impedance of ( a ) DU145, ( b ) MCF-7, ( c ) Jurkat, and ( d ) BV2 cells stained using four different protocols. The incubation time for calcein-stained cells was 20 min. Orange bars represent the acoustic impedance of unstained cells measured in parallel with the stained cells. All measurements were conducted between 1 and 3 hours after suspension in PBS solutions. Error bars indicate the fitting error of the mean obtained from fitting the data to the cumulative distribution function of a normal function. Brackets indicate contrasts (unstained and stained) tested with two-sided Wald z-tests on the two fitted estimates. Asterisks denote significance ( ∗ p < 0.05; ns = not significant). The p -values reflect within-experiment uncertainty of the fits.

Article Snippet: To examine the impact of staining on cell physical properties, four fluorescent dyes were used: a cell-permeable tracer calcein AM (Fisher Scientific International, Pittsburgh, PA) for cytoplasm staining; anti-EpCAM FITC (Becton, Dickinson and Company, Franklin Lakes, NJ), which binds to the cell membranes; E-cadherin CD324 PE (Miltenyi Biotec Norden, Lund, Sweden), also for cell membrane binding; and Hoechst 33342 (TargetMol Chemicals, Boston, MA), which binds to DNA in the cell nucleus.

Techniques: Staining, Incubation, Suspension

Journal: Biophysical Journal

Article Title: Impact of staining on cell acoustic properties

doi: 10.1016/j.bpj.2025.11.007

Figure Lengend Snippet: Measured acoustic impedance of calcein-stained MCF-7 cells at different incubation times. Orange bars represent the acoustic impedance of unstained cells measured in parallel with the stained cells. All measurements were conducted between 1 and 3 hours after suspension in PBS solutions. Error bars indicate the fitting error of the mean obtained from fitting the data to the cumulative distribution function of a normal function. Brackets indicate contrasts (unstained and stained) tested with two-sided Wald z-tests on the two fitted estimates. Asterisks denote significance ( ∗ p < 0.05; ∗∗ p < 0.01). The p -values reflect within-experiment uncertainty of the fits.

Article Snippet: To examine the impact of staining on cell physical properties, four fluorescent dyes were used: a cell-permeable tracer calcein AM (Fisher Scientific International, Pittsburgh, PA) for cytoplasm staining; anti-EpCAM FITC (Becton, Dickinson and Company, Franklin Lakes, NJ), which binds to the cell membranes; E-cadherin CD324 PE (Miltenyi Biotec Norden, Lund, Sweden), also for cell membrane binding; and Hoechst 33342 (TargetMol Chemicals, Boston, MA), which binds to DNA in the cell nucleus.

Techniques: Staining, Incubation, Suspension