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Three-dimensional analysis of endoplasmic reticulum structure based on processing of images captured by 2-photon and confocal microscopy

Project goals

We will test the applicability and efficiency of methods for estimating the length of ER tubules, their length density in cell, proportion of lamellar sheets in ER, mean size and size distribution of meshes in the ER net, characteristics of ER organization, anisotropy and texture. The quantitative evaluation of such characteristics together with ER three-dimensional reconstruction and visualization will provide a complex and objective information about its structure and thus will help in studies of ER changes, both in normally developing cells and cells suffering from various types of stress. The methods will be tested on GFP-transformed tobacco cells (line BY-2). The ER characteristics wil estimated by contemporary unbiased stereological methods and image analysis techniques applied to three-dimensional image data acquired by 2-photon and confocal microscopy.

Keywords

endoplasmic tericulum3-D analysis2-photopn microscopyconfocal microscopyimage analysisquantiative evaluationGFP-transformed tobacco cellsstereological methods

Public support

  • Provider

    Academy of Sciences of the Czech Republic

  • Programme

    The research grant projects for juniors

  • Call for proposals

    Juniorské badatelské grantové projekty 1 (SAV02003-XJ)

  • Main participants

    Fyziologický ústav AV ČR, v. v. i.
    Univerzita Karlova / Přírodovědecká fakulta

  • Contest type

    VS - Public tender

  • Contract ID

    —

Alternative language

  • Project name in Czech

    Trojrozměrná analýza struktury endoplazmatického retikula na základě zpracování obrazů z 2-fotonového a konfokálního mikroskopu

  • Annotation in Czech

    Budeme testovat použitelnost a účinnost metod pro měření délky tubulů endoplazmatického retikula (ER), jejich délkové hustoty v buňce, podílu lamelárních struktur v ER, průměrné velikosti a rozdělení velikostí ok v ER síti, charakteristik organizace, anizotropie a textury ER. Kvantitativní vyhodnocení těchto charakteristik spolu s trojrozměrnými rekonstrukcemi ER poskytne komplexní a objektivní informace o struktuře ER, což přispěje k podrobnějšímu studiu jeho změn jak v normálně se vyvíjejících buňkách, tak i v buňkách vystavených různým typům stresu. Tyto metody budou testovány na tabákových buňkách linie BY-2 transformovaných genem pro GFP. Charakteristiky ER budou odhadovány současnými stereologickými metodami a technikami analýzy obrazu aplikovanými na trojrozměrná obrazová data získaná 2-fotonovou a konfokální mikroskopií.

Scientific branches

  • R&D category

    ZV - Basic research

  • CEP classification - main branch

    EA - Morphology and cytology

  • CEP - secondary branch

    EB - Genetics and molecular biology

  • CEP - another secondary branch

    JC - Computer hardware and software

  • 10601 - Cell biology
    10602 - Biology (theoretical, mathematical, thermal, cryobiology, biological rhythm), Evolutionary biology
    10603 - Genetics and heredity (medical genetics to be 3)
    10604 - Reproductive biology (medical aspects to be 3)
    10605 - Developmental biology
    10608 - Biochemistry and molecular biology
    10609 - Biochemical research methods
    20206 - Computer hardware and architecture
    30101 - Human genetics

Completed project evaluation

  • Provider evaluation

    U - Uspěl podle zadání (s publikovanými či patentovanými výsledky atd.)

  • Project results evaluation

    Image analysis methods for measuring the length density of endoplasmic reticulum (ER) tubules in tobacco cells were developed and implemented. Changes in ER structure were found during early phases of toxic influence of Al ions and latrunculin B.

Solution timeline

  • Realization period - beginning

    Jan 1, 2003

  • Realization period - end

    Jan 1, 2005

  • Project status

    U - Finished project

  • Latest support payment

    —

Data delivery to CEP

  • Confidentiality

    S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

  • Data delivery code

    CEP06-AV0-KJ-U/07:5

  • Data delivery date

    Feb 12, 2014

Finance

  • Total approved costs

    1,767 thou. CZK

  • Public financial support

    1,186 thou. CZK

  • Other public sources

    0 thou. CZK

  • Non public and foreign sources

    0 thou. CZK

Basic information

Recognised costs

1 767 CZK thou.

Public support

1 186 CZK thou.

67%

Provider

Academy of Sciences of the Czech Republic

CEP

EA - Morphology and cytology

Solution period

01. 01. 2003 - 01. 01. 2005

Similar projects(10)

Segmentation, visualization and measurement of fibrous structures in 3D (MEB080831) 3-D studies of cells and their components (IAA5011810) Development of Reliable Methods for Automated Quantitative Characterization of Cell Motility in Fluorescence Microscopy (GJ16-03909Y)