For Research Use Only. Not for Diagnostic Use
MK-50 010 /Rev A/Draft Date 06192017
1
1.
One day before fixation, seed cells in growth
medium on chamber slides at a density that will
allow cells to be 80–90% confluent at the time of
fixation.
1.
Remove growth media, and dissemble chambers.
2.
Submerge the slides in a Tissue-Tek
®
container with
200 mL of 1X PBS.
IMPORTANT!
Do not let cells dry out at any time.
Always use enough solution to submerge all the
cells.
3.
Remove 1X PBS, and add 10% Neutral Buffered
Formalin (NBF). Incubate at
ROOM TEMPERATURE (RT)
for
30 MIN
.
4.
Remove NBF, and gently rinse slides with 1X
PBS.
Repeat twice.
1.
Remove 1X PBS wash, and replace with 50 mL 50%
EtOH. Incubate at
RT
for
5 MIN
.
2.
Remove 50% EtOH, and replace with 50 mL 70%
EtOH. Incubate at
RT
for
5 MIN
.
3.
Remove 70% EtOH, and replace with 50 mL 100%
EtOH. Incubate at
RT
for
5 MIN
.
4.
Remove 100% EtOH, and replace with fresh 100%
EtOH. Incubate at
RT
for
10 MIN
.
NOTE:
The slides can be stored in 100% EtOH
at
--20°C
for up to
6 MONTHS
.
1.
Submerge slides in 70% EtOH. Incubate at
RT
for
2 MIN
.
IMPORTANT!
Do not let cells dry out at any time.
Always use enough solution to submerge all the
cells.
2.
Remove 70% EtOH, and replace with 50% EtOH.
Incubate at
RT
for
2 MIN
.
3.
Remove 50% EtOH, and replace with 1X PBS.
Incubate at
RT
for
10 MIN
.