Determination of the oligomerization state of plasma membrane proteins and lipids is key to understanding cellular signaling and function. TOCCSL (thinning out clusters while conserving stoichiometry of labeling)—an imaging modality based on single-molecule fluorescence microscopy—allows for measuring the degree of homo- or hetero-oligomerization of membrane constituents even at expression levels......
A customized micropatterning approach combined with total internal reflection fluorescence microscopy was used to quantify ligand-induced FGFR3 receptor activation at the live cell membrane. Activation was recorded via the recruitment of the downstream signal transducer growth factor receptor-bound 2 (GRB2) to FGFR3 micropatterns. Authors: Ingrid Hartl 1, Veronika Brumovska 2, Yasmin Striedner 1, Atena Yasari 1, Gerhard J......
Using the single-molecule fluorescence microscopy modality “TOCCSL” and flux assays, the subunit stoichiometry and function of the norepinephrine transporter (NET) was investigated. NET was found to form monomers (~60%) and dimers (~40%) at the plasma membrane. The average oligomeric state was found to decrease upon PIP2 depletion. Authors: Dino Luethi 1 2, Julian Maier 1, Deborah......
Careful phenotype analysis of genetically altered mouse embryos/fetuses is vital for deciphering the function of pre- and perinatally lethal genes. Usually this involves comparing the anatomy of mutants with that of wild types of identical developmental stages. Detailed three dimensional information on regular cranial nerve (CN) anatomy of prenatal mice......
Magdalena C Schneider and Gerhard J Schütz describe how significance testing provides an important and valid addendum to the toolbox of quantitative (single molecule) biology. For two selected applications of single molecule microscopy – Nanoclustering in single molecule localization microscopy (SMLM) and single molecule tracking – they introduce how a single overall p-value......
Compared to room temperature experiments, SMLM performed under cryogenic temperature achieves higher photon yields and, hence, higher localization precision. However, to fully exploit the resolution it is crucial to account for the anisotropic emission characteristics of fluorescence dipole emitters with fixed orientation. We show here that astigmatic imaging in combination......
Approximately one-third of randomly produced knockout mouse lines produce homozygous offspring, which fail to survive the perinatal period. The majority of these die around or after embryonic day (E)14.5, presumably from cardiovascular insufficiency. For diagnosing structural abnormalities underlying death and diseases and for researching gene function, the phenotype of these......