frap analysis software  (Carl Zeiss)

Journal: Bioengineering & Translational Medicine

Article Title: Intratumoral nanofluidic system enhanced tumor biodistribution of PD‐L1 antibody in triple‐negative breast cancer

doi: 10.1002/btm2.10594

Figure Lengend Snippet: Tumor properties assessment of murine TNBC models, 4T1 and EMT6. Quantification of (a) cellular density ( n = 5), (b) αSMA fraction ( n = 4), and (c) collagen fraction ( n = 6) in 4T1 and EMT6 tumors non‐radiated and radiated. (d) Diffusion coefficients of 4T1 and EMT6 as determined via FRAP analysis ( n = 20 non‐radiated, n = 6 radiated). Ten ROIs were analyzed from three tumors of each TNBC model. Quantification of (e) CD45 + , (f) PD‐L1 + , and (g) CD45 + PD‐L1 + cells in tumor slides ( n = 5). Two‐tailed unpaired t ‐test was performed for statistical analysis. * p < 0.05; ** p < 0.005; n.s., no significance.

Article Snippet: FRAP analysis was performed using CellSens imaging software (Olympus).

Techniques: Diffusion-based Assay, Two Tailed Test

Journal: Chromosoma

Article Title: The effect of the lamin A and its mutants on nuclear structure, cell proliferation, protein stability, and mobility in embryonic cells

doi: 10.1007/s00412-016-0610-9

Figure Lengend Snippet: Analysis of HEK 293 sublines stably expressing EGFP alone and lamin A and its mutants D446V and Δ50 (progerin). a Confocal microscopy shows that all lamin A variants localize within the NE. Protein aggregates are much smaller than during transient transfection studies due to the lower level of exogenous proteins; however, some foci are still clearly visible. Progerin causes NE lobulations and the strongest deformation of the nuclear shape. b A proliferation comparison of the sublines was performed with the SRB assay 5 days after seeding. EGFP and lamin A expression significantly inhibits proliferation relative to the control, suggesting that the general effect of introducing exogenous protein may have a major role in this process. D446V and Δ50 abolished this effect, so they were able to increase the proliferation rate in embryonic cells. n/s not significant. c A comparison of protein mobility was performed using FRAP analysis. EGFP alone is very mobile: it immediately refilled the bleached area. By contrast, wild-type lamin A, progerin, and D446V proteins did not replace bleached lamin A. It shows that mutations did not significantly change protein mobility within the NE. Measurements were performed for up to 120 seconds. The insert nucleus picture shows the typical bleached area with a red circle (the merge of the EGFP and transmission light channels)

Article Snippet: FRAP analysis software (Zeiss LSM 510 software – ZEN 2008) was used for the experimental setup and data collection (pre-bleach imaging, laser pulse, recovery imaging, collection of data, etc.).

Techniques: Stable Transfection, Expressing, Confocal Microscopy, Transfection, Comparison, Sulforhodamine B Assay, Control, Transmission Assay