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