Remove gelatin chamber and Lowicryl can. Mop the cryochamber floor
and temperature tube top with a Kimwipe so no Lowicryl puddles remain.
Replace the black temperature tube with the red one.
Close cover, lower and release glass plate, re-open cover and insert UV
lamp apparatus, connect to AFS and plug into power. Turn on the UV
power switch. The UV indicator light on control panel should show a
steady light. If it is flashing, the UV light is not lit. Check connections and
seating of apparatus.
Days 6, 7: No procedures
Day 8
Materials and Equipment:
Acetone for cleaning
Styrofoam box, foil-lined, with UV lamp (BLAK-RAY Lamp, longwave
UV366 nm, UVP, Upland, CA) embedded in lid to shine into box, and
raised interior platform, or commercial chillable UV curing box
Dry ice
Procedures:
To ensure that capsules are sufficiently cured, set up Styrofoam UV box
with dry ice and platform at about 2″ away from UV lamp (or use commercial box).
Remove curing cans from AFS, remove spider covers with attached capsules (labeling each set), blotting dry on paper towel, making sure capsules
are stable.
Set the spider covers on the platform with the capsules facing up.
Replace the UV lid and turn on.
Let cure overnight.
When curing is complete, remove one capsule at a time, noting position,
slit the gelatin and Flo-through capsule with a razor blade and peel them
away from the Lowicryl block; label block if no label was inserted.
Repeat for remaining capsules.
Empty ethanol from curing cans (dispose as hazardous waste).
Place cans with stem holders and bottom spacer plates in acetone for
cleaning, rinsing off residual resin debris, then air dry.
Spider covers should have cured resin residue gently removed with a
dissecting needle, then clean in acetone and air dry.
6 Conjugate Immunofluorescence—SEM Array Tomography …
173
and temperature tube top with a Kimwipe so no Lowicryl puddles remain.
Replace the black temperature tube with the red one.
Close cover, lower and release glass plate, re-open cover and insert UV
lamp apparatus, connect to AFS and plug into power. Turn on the UV
power switch. The UV indicator light on control panel should show a
steady light. If it is flashing, the UV light is not lit. Check connections and
seating of apparatus.
Days 6, 7: No procedures
Day 8
Materials and Equipment:
Acetone for cleaning
Styrofoam box, foil-lined, with UV lamp (BLAK-RAY Lamp, longwave
UV366 nm, UVP, Upland, CA) embedded in lid to shine into box, and
raised interior platform, or commercial chillable UV curing box
Dry ice
Procedures:
To ensure that capsules are sufficiently cured, set up Styrofoam UV box
with dry ice and platform at about 2″ away from UV lamp (or use commercial box).
Remove curing cans from AFS, remove spider covers with attached capsules (labeling each set), blotting dry on paper towel, making sure capsules
are stable.
Set the spider covers on the platform with the capsules facing up.
Replace the UV lid and turn on.
Let cure overnight.
When curing is complete, remove one capsule at a time, noting position,
slit the gelatin and Flo-through capsule with a razor blade and peel them
away from the Lowicryl block; label block if no label was inserted.
Repeat for remaining capsules.
Empty ethanol from curing cans (dispose as hazardous waste).
Place cans with stem holders and bottom spacer plates in acetone for
cleaning, rinsing off residual resin debris, then air dry.
Spider covers should have cured resin residue gently removed with a
dissecting needle, then clean in acetone and air dry.
6 Conjugate Immunofluorescence—SEM Array Tomography …
173