Evidence map›Paper›PMID 25786567›Full record

ArticleJournal of microscopy2015

Preservation of protein fluorescence in embedded human dendritic cells for targeted 3D light and electron microscopy.

K Höhn, J Fuchs, A Fröber, R Kirmse, B Glass, M Anders-Össwein, P Walther, H-G Kräusslich, C Dietrich

Abstract read
In one paragraph

Article in Journal of microscopy, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

3 citing papers in PubMed.

  1. Article
  2. Review
  3. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

9 authors.

K HöhnDepartment of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany.
J FuchsCarl Zeiss AG, Oberkochen, Germany.
A FröberCarl Zeiss AG, Oberkochen, Germany.
R KirmseCarl Zeiss Microscopy GmbH, Jena, Germany.
B GlassDepartment of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany.
M Anders-ÖssweinDepartment of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany.
P WaltherElectron Microscopy Facility, Ulm University, Ulm, Germany.
H-G KräusslichDepartment of Infectious Diseases, Virology, University Hospital Heidelberg, Heidelberg, Germany.
C DietrichCarl Zeiss AG, Oberkochen, Germany.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

In this study, we present a correlative microscopy workflow to combine detailed 3D fluorescence light microscopy data with ultrastructural information gained by 3D focused ion beam assisted scanning electron microscopy. The workflow is based on an optimized high pressure freezing/freeze substitution protocol that preserves good ultrastructural detail along with retaining the fluorescence signal in the resin embedded specimens. Consequently, cellular structures of interest can readily be identified and imaged by state of the art 3D confocal fluorescence microscopy and are precisely referenced with respect to an imprinted coordinate system on the surface of the resin block. This allows precise guidance of the focused ion beam assisted scanning electron microscopy and limits the volume to be imaged to the structure of interest. This, in turn, minimizes the total acquisition time necessary to conduct the time consuming ultrastructural scanning electron microscope imaging while eliminating the risk to miss parts of the target structure. We illustrate the value of this workflow for targeting virus compartments, which are formed in HIV-pulsed mature human dendritic cells.

Indexed as

Cells, CulturedDendritic CellsFluorescenceFreeze SubstitutionFreezinggag Gene Products, Human Immunodeficiency VirusHIVHumansImaging, Three-DimensionalLuminescent ProteinsMicroscopy, ConfocalMicroscopy, ElectronMicroscopy, Electron, ScanningMicroscopy, FluorescenceMicrotomyRecombinant Proteinsgag Gene Products, Human Immunodeficiency VirusLuminescent ProteinsRecombinant ProteinsRed Fluorescent ProteinConfocal laser scanning microscopycorrelative light and electron microscopyfluorescence preservation of embedded samplesfocused ion beamtargeted FIB

Identifiers

PMID25786567
PMCPMC4757415

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.