human foreskin fibroblasts cells hff 1 scrc 1041  (ATCC)

Journal: Langmuir

Article Title: Traction Force Microscopy with DNA FluoroCubes

doi: 10.1021/acs.langmuir.5c04971

Figure Lengend Snippet: Demonstration of TFM with FluoroCubes and fluorescent beads as fiducial markers. (a) (i) and (ii) bright-field image of a murine kidney fibroblast with GFP-labeled kindlin-2 (Kind Ko+K2GFP cell line) and the corresponding fluorescent image; (b) displacement correlation between the bead and FluoroCube channel resulting from the stage shift in the x direction. The black dots represent the shifts measured by the conventional KLT method from the FluoroCube channel as a function of the corresponding shifts in the bead channel. The fitted curve has a slope of 0.9975 with R 2 = 0.9940; (c) (i) and (ii) displacement magnitudes measured in the bead and FluoroCube channels, respectively; (d) correlation of the displacements shown in (c) (i,ii); the micrometer-to-pixel factor is 0.103 μm/pix; (e) (i) traction forces calculated from the displacements in (c) (i) with BFTTC; (e) (ii) traction forces calculated from the displacements in (c) (ii); (e) (iii) traction forces calculated from dual-channel optical flow tracking results (see Figure S7 ). All scale bars represent 12.5 μm.

Article Snippet: Human foreskin fibroblasts (ATCC; SCRC-1041) and Kind Ko+K2GFP cells (Kind Ko immortalized murine kidney fibroblast reconstituted with green fluorescent protein-tagged kindlin-2 expression plasmids, generous gift from Prof. Dr. Reinhard Fässler, PMID: 26821125) were cultured at 37 °C and in 5% CO 2 in Dulbecco’s modified Eagle’s medium (DMEM; Sigma-Aldrich, D6546), supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich, F2442), 4 mM l -glutamine (Sigma-Aldrich, G7513), and 20 μg/mL gentamicin (Sigma-Aldrich, G1397).

Techniques: Labeling