Correlative Microscopy of Ultrathin Cryosections is a Powerful Tool for Placental Research

Placenta ◽  
2003 ◽  
Vol 24 (5) ◽  
pp. 557-565 ◽  
Author(s):  
T. Takizawa ◽  
J.M. Robinson
Keyword(s):  
Correlative Microscopy ◽  
Ultrathin Cryosections

scholarly journals Correlative Microscopy Using FluoroNanogold on Ultrathin Cryosections: Proof of Principle

1998 ◽  
Vol 46 (10) ◽  
pp. 1097-1102 ◽  
Author(s):  
Toshihiro Takizawa ◽  
Kouki Suzuki ◽  
John M. Robinson
Keyword(s):  
Electron Microscopy ◽  
Imaging Techniques ◽  
Human Neutrophils ◽  
Fluorescence Signal ◽  
Correlative Microscopy ◽  
Reporter System ◽  
Fluorescence And Electron Microscopy ◽  
Specific Granules ◽  
Ultrathin Cryosections ◽  
Proof Of Principle

We demonstrate a fluorescent ultrasmall immunogold probe, FluoroNanogold (FNG), to be a versatile reporter system for immunocytochemical labeling of ultrathin cryosections. FNG-labeled molecules in the same ultrathin cryosections can be resolved by two imaging techniques (i.e., fluorescence and electron microscopy). Lactoferrin, a marker protein for the specific granules in human neutrophils, was employed as the target for FNG immunolabeling. The spatial resolution of the fluorescence signal from FNG-labeled specific granules was compatible with that of silver-enhanced gold signal from the same granules in electron microscopy. Our results confirm that FNG can be used as a probe for highresolution correlation between immunofluorescence and electron microscopy.

Correlative microscopy in distrubuted (client/server) computing environments: tools for catalyzing the rapid dissemination of information about the cell biology of the human prostate

1995 ◽  
Vol 53 ◽  
pp. 682-683
Author(s):  
A. Hakam ◽  
J.T. Gau ◽  
M.L. Grove ◽  
B.A. Evans ◽  
M. Shuman ◽  
...  
Keyword(s):  
United States ◽  
Cell Biology ◽  
Tissue Bank ◽  
The United States ◽  
Prostate Adenocarcinoma ◽  
Correlative Microscopy ◽  
Client Server ◽  
Critical Elements ◽  
Transplant Organ

Prostate adenocarcinoma is the most common malignant tumor of men in the United States and is the third leading cause of death in men. Despite attempts at early detection, there will be 244,000 new cases and 44,000 deaths from the disease in the United States in 1995. Therapeutic progress against this disease is hindered by an incomplete understanding of prostate epithelial cell biology, the availability of human tissues for in vitro experimentation, slow dissemination of information between prostate cancer research teams and the increasing pressure to “ stretch” research dollars at the same time staff reductions are occurring.To meet these challenges, we have used the correlative microscopy (CM) and client/server (C/S) computing to increase productivity while decreasing costs. Critical elements of our program are as follows:1) Establishing the Western Pennsylvania Genitourinary (GU) Tissue Bank which includes >100 prostates from patients with prostate adenocarcinoma as well as >20 normal prostates from transplant organ donors.

High-resolution SEM and STEM of adrenocortical endothelium: Molecular resolution of membrane complexes

1993 ◽  
Vol 51 ◽  
pp. 610-611
Author(s):  
Robert P. Apkarian
Keyword(s):  
Fine Structure ◽  
Low Voltage ◽  
Uranyl Acetate ◽  
Correlative Microscopy ◽  
Coated Pits ◽  
Fine Grain ◽  
Structural Identity ◽  
Cellular Pathways ◽  
Steroidogenic Cells ◽  
Cr Film

A multitude of complex ultrastructural features are involved in endothelial cell (EC) gating and sorting of lipid through capillaries and into steroidogenic cells of the adrenal cortex. Correlative microscopy is necessary to distinguish the structural identity of features involved in specific cellular pathways. In addition to diaphragmed fenestrae that frequently appear in clusters, other 60-80 nm openings; plasmalemma vesicles (PV), channels and pockets fitted with diaphragms of the same dimension, coexist on the thin EC surface. Non-diaphragmed coated pits (CP) (100-120 nm) involved in receptor mediated endocytosis were also present on the EC membrane. The present study employed HRSEM of cryofractured and chromium coated specimens and low voltage HRSTEM of 80 nm thick LX-112 embedded sections stained with 2.0% uranyl acetate. Both preparations were imaged at 25 kV with a Topcon DS-130 FESEM equipped with in-lens stage and STEM detector.HRSEM images of the capillary lumen coated with a lnm continuous fine grain Cr film, provided the ability to scan many openings and resolve (SE-I contrast) the fine structure of diaphragm spokes and central densities (Fig. 1).

Improved immunolabelling of ultrathin cryosections using antibody conjugated with 1nm gold particles

1990 ◽  
Vol 48 (3) ◽  
pp. 928-929
Author(s):  
Morten H. Nielsen ◽  
Lone Bastholm
Keyword(s):  
Pituitary Gland ◽  
Liquid Nitrogen ◽  
Electron Density ◽  
Small Diameter ◽  
Cytochemical Staining ◽  
Gold Particles ◽  
Labelling Density ◽  
Ultrathin Cryosections ◽  
Mouse Pituitary ◽  
Electron Microscopical

During the last 5 years the diameter of the gold probes used for immuno-cytochemical staining at the electron microscopical (EM) level has been decreased. The advantage of small diameter gold probes is an overall increased labelling density. The disadvantage is a lower detectability due to the low electron density of smaller gold particles consequently an inconvenient high primary magnification needed for EM examination. Since 1 nm gold particles are barely visible by conventional EM examination the need for enlargement by silverenhancement of the gold particles has increased.In the present study of ultrathin cryosectioned material the results of immunostaining using 5 nm gold conjugated antibody and 1 nm gold conjugated antibodies are compared after silverenhancement of the 1 nm gold particles.Slices of freshly isolated mouse pituitary gland were immersion fixed for 20 min in 2 % glutaraldehyde /2 % paraformaldehyde. Blocks cryoprotected with 2.3 M sucrose were frozen in liquid nitrogen and ultra-cryosectioned on a RMC cryoultra-microtome.

Phase Characterization of a Biocompatible Co-Cr-W Alloy Via Correlative Microscopy

2016 ◽  
Vol 53 (7) ◽  
pp. 450-461 ◽  
Author(s):  
I. Weißensteiner ◽  
P. Voigt ◽  
V. Maier-Kiener ◽  
H. Clemens
Keyword(s):  
Correlative Microscopy ◽  
Phase Characterization

Structured illumination microscopy and correlative microscopy to study autophagy

Methods ◽  
2015 ◽  
Vol 75 ◽  
pp. 61-68 ◽  
Author(s):  
Laure-Anne Ligeon ◽  
Nicolas Barois ◽  
Elisabeth Werkmeister ◽  
Antonino Bongiovanni ◽  
Frank Lafont
Keyword(s):  
Structured Illumination ◽  
Correlative Microscopy ◽  
Structured Illumination Microscopy

scholarly journals Revealing Inflammatory Indications Induced by Titanium Alloy Wear Debris in Periprosthetic Tissue by Label-Free Correlative High-Resolution Ion, Electron and Optical Microspectroscopy

Materials ◽  
2021 ◽  
Vol 14 (11) ◽  
pp. 3048
Author(s):  
Rok Podlipec ◽  
Esther Punzón-Quijorna ◽  
Luka Pirker ◽  
Mitja Kelemen ◽  
Primož Vavpetič ◽  
...  
Keyword(s):  
Titanium Alloy ◽  
High Resolution ◽  
Wear Debris ◽  
Ion Beam ◽  
Quantitative Measure ◽  
Label Free ◽  
Periprosthetic Tissue ◽  
Correlative Microscopy ◽  
X Ray ◽  
Tissue Reactions

The metallic-associated adverse local tissue reactions (ALTR) and events accompanying worn-broken implant materials are still poorly understood on the subcellular and molecular level. Current immunohistochemical techniques lack spatial resolution and chemical sensitivity to investigate causal relations between material and biological response on submicron and even nanoscale. In our study, new insights of titanium alloy debris-tissue interaction were revealed by the implementation of label-free high-resolution correlative microscopy approaches. We have successfully characterized its chemical and biological impact on the periprosthetic tissue obtained at revision surgery of a fractured titanium-alloy modular neck of a patient with hip osteoarthritis. We applied a combination of photon, electron and ion beam micro-spectroscopy techniques, including hybrid optical fluorescence and reflectance micro-spectroscopy, scanning electron microscopy (SEM), Energy-dispersive X-ray Spectroscopy (EDS), helium ion microscopy (HIM) and micro-particle-induced X-ray emission (micro-PIXE). Micron-sized wear debris were found as the main cause of the tissue oxidative stress exhibited through lipopigments accumulation in the nearby lysosome. This may explain the indications of chronic inflammation from prior histologic examination. Furthermore, insights on extensive fretting and corrosion of the debris on nm scale and a quantitative measure of significant Al and V release into the tissue together with hydroxyapatite-like layer formation particularly bound to the regions with the highest Al content were revealed. The functional and structural information obtained at molecular and subcellular level contributes to a better understanding of the macroscopic inflammatory processes observed in the tissue level. The established label-free correlative microscopy approach can efficiently be adopted to study any other clinical cases related to ALTR.

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