Scribd Logo
Upload

Welcome to Scribd!

  • 36 views

    Photos PDF

    Uploaded by

    rakeshrakesh
    The document describes techniques used to isolate organelles from cells for study under an electron microscope. Cells are placed in a buffered isotonic solution and broken open to release organelles. The solution is filtered and ultracentrifuged at increasing speeds to separate organelles by weight, allowing isolation of heavier components like nuclei followed by smaller fragments. This process produces large numbers of purified organelles that can then be examined individually using electron microscopy.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd

    Photos PDF

    Uploaded by

    rakeshrakesh
    36 views4 pages
    The document describes techniques used to isolate organelles from cells for study under an electron microscope. Cells are placed in a buffered isotonic solution and broken open to release organelles. The solution is filtered and ultracentrifuged at increasing speeds to separate organelles by weight, allowing isolation of heavier components like nuclei followed by smaller fragments. This process produces large numbers of purified organelles that can then be examined individually using electron microscopy.

    Original Title

    photos.pdf

    Copyright

    © © All Rights Reserved

    Available Formats

    PDF, TXT or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    The document describes techniques used to isolate organelles from cells for study under an electron microscope. Cells are placed in a buffered isotonic solution and broken open to release organelles. The solution is filtered and ultracentrifuged at increasing speeds to separate organelles by weight, allowing isolation of heavier components like nuclei followed by smaller fragments. This process produces large numbers of purified organelles that can then be examined individually using electron microscopy.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    36 views4 pages

    Photos PDF

    Uploaded by

    rakeshrakesh
    The document describes techniques used to isolate organelles from cells for study under an electron microscope. Cells are placed in a buffered isotonic solution and broken open to release organelles. The solution is filtered and ultracentrifuged at increasing speeds to separate organelles by weight, allowing isolation of heavier components like nuclei followed by smaller fragments. This process produces large numbers of purified organelles that can then be examined individually using electron microscopy.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    You are on page 1of 4

    Transparency:

    Features of cells visible using an electron microscope (1)


    cell surface membrane
    of upper cell

    texture of cell wall


    (in plant cells only)

    cell wall

    cell surface membrane


    of lower cell

    cytoplasm

    plasmodesma
    (25 nm wide)

    micrograph of cell wall


    granum
    (with chlorophyll)

    membrane
    lipid (fat)

    starch outer membrane of


    grain chloroplast envelope

    (lamella)

    inner membrane of stroma


    chloroplast envelope

    micrograph of chloroplast (x 8000)


    D. Ehrig, Brinkum
    organelles_em

    Transparency:

    Features of cells visible using an electron microscope (2)


    nucleolus

    chromatin (= disorganized genetic material)

    nuclear
    pore

    outer
    inner
    membrane

    membrane of cell
    surface (x 100000).
    At this magnification it appears
    as two (!) dark
    lines at the edge
    of the cell.

    micrograph of nucleus

    outer membrane

    inner membrane

    matrix

    cristae (sing. crista)

    lipid

    micrograph of mitochondrion (x 40000)


    D. Ehrig, Brinkum
    organelles_em

    Transparency:

    Features of cells visible using an electron microscope (3)


    cell surface membrane
    secretory vesicles
    Golgi apparatus
    vesicle adding to and
    budding from pile of
    membranes
    Electron micrograph
    of Golgi body
    (apparatus)

    microtubes, some of
    which are transporting
    secretory vesicles to the
    cell surface
    Golgi apparatus (x 72500)

    Electron micrograph of free ribosomes


    Electron micrograph of
    endoplasmic reticulum
    (x 55000)

    Electron micrograph of
    rough endoplasmic
    reticulum
    (ER + bound ribosomes)
    D. Ehrig, Brinkum
    organelles_em

    Transparency:

    Features of cells visible using an electron microscope (4)

    mitochondrion
    cytoplasm
    nucleus
    cell wall

    large vacuole
    cell surface
    membrane

    chloroplast

    tonoplast

    Appearance of a
    representative plant
    cell as seen with an
    electron microscope
    (x 12000)

    Cell fractionation and ultracentrifugation


    In order to study the structure and function of the various organelles which make up cells, it is
    necessary to obtain [erhalten, bekommen] large numbers of isolated organelles. There are two stages in achieving [erreichen] this:
    Cell fractionation involves [beinhalten, bedeuten] cells being placed in a cold, isotonic, buffered
    solution, for the following reasons:
    - cold - to reduce enzyme activity which might break down the organelles
    - isotonic [isotonisch = gleiche Konzentration innerhalb und auerhalb der Organelle] - to prevent
    organelles bursting due to the influx [Einflieen] of water
    - buffered [gepuffert] - to maintain a constant pH [Suregrad].
    They are then broken up using a pestle [Stsel, Pistill] and mortar [Mrser] or an electrical blender
    [Mixer/Zerkleinerer] to break the cell membrane and/or wall and release [freisetzen] the organelles.
    The resultant fluid is then filtered to remove any complete cells and large pieces of debris
    [Reststcke, Mll].
    Ultracentrifugation is the process by which fragments in the filtered
    liquid are separated in a machine called a centrifuge. This spins tubes
    of the liquid at very high speed, to create a centrifugal force. At slower
    speeds, only the very heaviest organelles are forced out of suspension,
    into a thin deposit [Ablagerung] at he bottom of the tube. This deposit
    includes the nuclei, which can be isolated by removing the supernatant
    liquid [berstand(-flssigkeit)]. This supernatant liquid can then be spun in
    the centrifuge at a faster speed, separating out the next most heavy
    component [Bestandteil]. By continuing in this way, smaller and smaller
    fragments can be separated out.
    D. Ehrig, Brinkum
    organelles_em

    You might also like