quantitative speckle fluorescence microscopy (qfsm) software  (MathWorks Inc)

Journal: Biology Open

Article Title: The Drosophila protein, Nausicaa, regulates lamellipodial actin dynamics in a Cortactin-dependent manner

doi: 10.1242/bio.038232

Figure Lengend Snippet: Nau s regulates lamellipodial actin density and actin retrograde flow in S2R+ cells. (A,B) Fixed S2R+ cells treated with control (A) or Naus RNAi (B) stained for F-actin with phalloidin. Gray levels have been set equal for comparison. Scale bar: 10 µm. (C) Line-scan analysis of F-actin fluorescence in the lamellipodia from cells as shown in A and B. Fluorescence was normalized for each cell and averaged for each condition (black circles, control RNAi and open gray, circles Naus RNAi). (D) Mean fluorescence intensity of lamellipodial actin of cells treated with control RNAi (black circles) or Naus RNAi (open black circles) from cells as shown in A and B ( P <0.0001, Student's t -test, n =30 cells per condition). (E,F) Representative heat maps of actin speeds in (E) control or (F) Naus RNAi-treated cells from QFSM analysis. Cool colors indicate slower rates of retrograde flow and warm colors represent faster speeds of actin retrograde flow. Scale bar: 10 µm. (G) Quantification of lamellipodial actin speeds from QFSM analysis ( P= 0.0059, Mann–Whitney test, control RNAi: n =40 cells, Naus RNAi: n =46 cells). (H) Quantification of the mean fluorescence intensity of EGFP-Actin in the cells analyzed by QFSM (black circles, control RNAi and open black circles, Naus RNAi), n.s., not significant (Mann–Whitney test).

Article Snippet: The resulting movies were analyzed using a previously described Quantitative Fluorescent Speckle Microscopy (QFSM) software in MATLAB ( ).

Techniques: Control, Staining, Comparison, Fluorescence, MANN-WHITNEY