Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination.
AuthorsGustafsson MG, Shao L, Carlton PM, Wang CJ, Golubovskaya IN, Cande WZ, Agard DA, Sedat JW,
JournalBiophys J
PubMed ID18326650
'Structured illumination microscopy is a method that can increase the spatial resolution of wide-field fluorescence microscopy beyond its classical limit by using spatially structured illumination light. Here we describe how this method can be applied in three dimensions to double the axial as well as the lateral resolution, with true ... More
Mapping mechanical strain of an endogenous cytoskeletal network in living endothelial cells.
AuthorsHelmke BP, Rosen AB, Davies PF
JournalBiophys J
PubMed ID12668477
'A central aspect of cellular mechanochemical signaling is a change of cytoskeletal tension upon the imposition of exogenous forces. Here we report measurements of the spatiotemporal distribution of mechanical strain in the intermediate filament cytoskeleton of endothelial cells computed from the relative displacement of endogenous green fluorescent protein (GFP)-vimentin before ... More
a2 Adrenergic receptor-mediated inhibition of thermogenesis.
AuthorsMadden CJ, Tupone D, Cano G, Morrison SF,
JournalJ Neurosci
PubMed ID23365239
'a2 adrenergic receptor (a2-AR) agonists have been used as antihypertensive agents, in the management of drug withdrawal, and as sedative analgesics. Since a2-AR agonists also influence the regulation of body temperature, we explored their potential as antipyretic agents. This study delineates the central neural substrate for the inhibition of rat ... More
Glucoprivation in the ventrolateral medulla decreases brown adipose tissue sympathetic nerve activity by decreasing the activity of neurons in raphe pallidus.
AuthorsMadden CJ,
JournalAm J Physiol Regul Integr Comp Physiol
PubMed ID22071154
'In urethane/a-chloralose anesthetized rats, cold exposure increased brown adipose tissue sympathetic nerve activity (BAT SNA: +699 ± 104% control). Intravenous administration of 2-deoxy-D-glucose (2-DG; 200 mg·ml(-1)·kg(-1)) reversed the cold-evoked activation of BAT SNA (nadir: 139 ± 36% of control) and decreased BAT temperature (-1.1 ± 0.2°C), expired CO(2) (-0.4 ± ... More
Spatiotemporal analysis of flow-induced intermediate filament displacement in living endothelial cells.
'The distribution of hemodynamic shear stress throughout the arterial tree is transduced by the endothelium into local cellular responses that regulate vasoactivity, vessel wall remodeling, and atherogenesis. Although the exact mechanisms of mechanotransduction remain unknown, the endothelial cytoskeleton has been implicated in transmitting extracellular force to cytoplasmic sites of signal ... More
Multicolor super-resolution imaging with photo-switchable fluorescent probes.
AuthorsBates M, Huang B, Dempsey GT, Zhuang X,
JournalScience
PubMed ID17702910
Recent advances in far-field optical nanoscopy have enabled fluorescence imaging with a spatial resolution of 20 to 50 nanometers. Multicolor super-resolution imaging, however, remains a challenging task. Here, we introduce a family of photo-switchable fluorescent probes and demonstrate multicolor stochastic optical reconstruction microscopy (STORM). Each probe consists of a photo-switchable ... More
I5S: wide-field light microscopy with 100-nm-scale resolution in three dimensions.
AuthorsShao L, Isaac B, Uzawa S, Agard DA, Sedat JW, Gustafsson MG,
JournalBiophys J
PubMed ID18326649
A new type of wide-field fluorescence microscopy is described, which produces 100-nm-scale spatial resolution in all three dimensions, by using structured illumination in a microscope that has two opposing objective lenses. Illumination light is split by a grating and a beam splitter into six mutually coherent beams, three of which ... More
Central activation of the A1 adenosine receptor (A1AR) induces a hypothermic, torpor-like state in the rat.
AuthorsTupone D, Madden CJ, Morrison SF,
Journal
PubMed ID24005302
Since central activation of A1 adenosine receptors (A1ARs) plays an important role in the induction of the hypothermic and hypometabolic torpid state in hibernating mammals, we investigated the potential for the A1AR agonist N6-cyclohexyladenosine to induce a hypothermic, torpor-like state in the (nonhibernating) rat. Core and brown adipose tissue temperatures, ... More
Inhibition of brown adipose tissue thermogenesis by neurons in the ventrolateral medulla and in the nucleus tractus solitarius.
AuthorsCao WH, Madden CJ, Morrison SF,
JournalAm J Physiol Regul Integr Comp Physiol
PubMed ID20410479
Neurons in the ventrolateral medulla (VLM) and in the nucleus tractus solitarius (NTS) play important roles in the regulation of cardiovascular and other autonomic functions. In the present study, we demonstrate an inhibition of brown adipose tissue (BAT) thermogenesis evoked by activation of neurons in the VLM, as well as ... More
Endogenous activation of spinal 5-hydroxytryptamine (5-HT) receptors contributes to the thermoregulatory activation of brown adipose tissue.
AuthorsMadden CJ, Morrison SF,
JournalAm J Physiol Regul Integr Comp Physiol
PubMed ID20071609
Neurons in the rostral raphe pallidus (RPa) play an essential role in the regulation of sympathetically mediated metabolism and thermogenesis in brown adipose tissue (BAT). The presence of serotonergic neurons in the RPa that are retrogradely labeled following pseudorabies virus injections into BAT suggests that these neurons play a role ... More
Translational diffusion of fluorescent proteins by molecular fourier imaging correlation spectroscopy.
AuthorsFink MC, Adair KV, Guenza MG, Marcus AH
JournalBiophys J
PubMed ID16920833
The ability to noninvasively observe translational diffusion of proteins and protein complexes is important to many biophysical problems. We report high signal/noise (>or=250) measurements of the translational diffusion in viscous solution of the fluorescent protein, DsRed. This is carried out using a new technique: molecular Fourier imaging correlation spectroscopy (M-FICS). ... More
Measurement of monomer-oligomer distributions via fluorescence moment image analysis.
AuthorsSergeev M, Costantino S, Wiseman PW
JournalBiophys J
PubMed ID16935950
We present higher-order moment analysis of fluorescence intensity fluctuations from individual laser scanning microscopy images applied to study monomer-oligomer distributions. We demonstrate that the number densities and brightness ratios of a mixed population of monomers and oligomers can be determined by analyzing higher-order moments of the fluorescence intensity fluctuations from ... More
In situ fluorescent protein imaging with metal film-enhanced total internal reflection microscopy.
AuthorsBurghardt TP, Charlesworth JE, Halstead MF, Tarara JE, Ajtai K
JournalBiophys J
PubMed ID16565065
Fluorescence detection of single molecules provides a means to investigate protein dynamics minus ambiguities introduced by ensemble averages of unsynchronized protein movement or of protein movement mimicking a local symmetry. For proteins in a biological assembly, taking advantage of the single molecule approach could require single protein isolation from within ... More
Adenovirus-mediated gene transfer to human bronchial submucosal glands using xenografts.
AuthorsPilewski JM, Engelhardt JF, Bavaria JE, Kaiser LR, Wilson JM, Albelda SM
JournalAm J Physiol
PubMed ID7733306
The cystic fibrosis (CF) transmembrane conductance regulator has been localized to both submucosal glands and surface epithelium, suggesting that both glandular and surface epithelium may be important targets for gene therapy. To determine the distribution and efficiency of recombinant adenovirus-mediated gene transfer to human airway submucosal glands, an in vivo ... More