Tehnici de centrifugure Tehnici de centrifugure

For For|e core oc|ioneo; osupro unei |e core oc|ioneo; osupro unei
porficuIe n procesuI de porficuIe n procesuI de
sedimenfore sedimenfore::
2 Iorte:
- greutatea, G,
i
- Forta de
Irecare, F
f
G
F
Gmg
F6rqv
m masa particulei,
g acceleratia
gravitational,
r raza particulei,
coeIicientul de
viscozitate,
v viteza particulei
n miscare
F
f
C
4r]u 4r]u centrifug centrifugdd - -
r r4tu 4tu]iu ]iu pp4ute 4ute m4difi m4dificu cu
urtifici urtificiuI uI S: S:
F
f
C
prubeta cu proba
in procesul de
sedimenatre
F
f
F
c
F
c
Eprubeta cu proba n rotajie
Forja centrifug
F
f
C
# ' # mc F
2 2
c
L
unde.
e 264 vite:a rotational
m masa particulei
8 et 8 densittile particulei i respectiv a solventului
' volumul particulei
4 frecventa de rotatie
# distanta ra:a centrifugei
uct uct44r ruI uI d de e eficien eficien]d ]d
uu centrifuge centrifugei i::

g
#

mg
# mc

G
Fc
I
2
2
2
. g)
Dispozitivul Dispozitivul
W Exemplu de separare prin centrifugare n
gradient de densitate
Tehn Tehnici ici de centrifuga de centrifugare re
ULTRACENTRIUSAREA
$;edberg - I9Z3.
!ono Io I30 000 - Ib0 000 rpm
,I 000 000g
$isfem de rocire
UItrucentrifugure z4nuIu
in grudient de densitute
tape:
1. Prelevarea
2. Omogenizarea
3. Sedimentarea
4. Centrifugarea
diferenial
Fractio Fractionarea narea celular celular
Tissue
cells
Homogenization
Homogenate 1000
1000 times the
force of gravity)
10 min ifferentiaI centrifugation
$upernatant poured
into next tube
20,000
20 min
Pellet rich in
nuclei and
cellular debris
Pellet rich in
mitochondria
and chloro-
plasts if cells
are from a
plant)
Pellet rich in
"microsomes
pieces of
plasma mem-
branes and
cells' internal
membranes)
Pellet rich in
ribosomes
150,000
3 hr
80,000
60 min
1. Vitez mic
(1.000gX 10 min):
celule ntregi,
nuclee, citoschelet
2. Vitez medie
(20.000gX 20
min): mitocondrii
lizozomi
3. Vitez mare
(80.000gX 1 h):
microzomi
4. Ultracentrifugare
(150.000gX 3 h):
ribozomi
macromolecule.
Centrifugureu
diferen]iuId