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Rhodamine-Conjugated Chitin Probe
From 2010.igem.org
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#* ''To prevent stain from evaporating overnight'': Use a large glass petri dish. Soak a paper towel with PBS and place it at the bottom of the petri dish. Use 2 toothpicks to support the slide above the paper towel. This prevents any wicking action from occurring and pulling the stain off the slide. | #* ''To prevent stain from evaporating overnight'': Use a large glass petri dish. Soak a paper towel with PBS and place it at the bottom of the petri dish. Use 2 toothpicks to support the slide above the paper towel. This prevents any wicking action from occurring and pulling the stain off the slide. | ||
# Wash 3 times with 1X TBS (submerge in 1X TBS for 5 minutes per wash) | # Wash 3 times with 1X TBS (submerge in 1X TBS for 5 minutes per wash) | ||
| + | # Place a cover slide over sample. Image sample using fluorescent microscope | ||
Latest revision as of 19:32, 14 August 2010
Methanol Fixation
- Rinse slide with ethanol and flame
- Rinse slide with PBS and dry with KimWipe
- Apply cell sample to microscope slide and let air dry
- Submerge in -20C absolute methanol for 5-10 min
- Wash 3 times with 1X TBS (submerge in 1X TBS for 5 minutes per wash)
- Do not let cells dry for the rest of the procedure
Staining
- Make 1:500 dilution of chitin probe in TBS (50ul of probe is used per slide, [49.9ul of 1X TBS]:[0.1ul of chitin probe] --> 1 slide)
- Apply 50ul to the slide containing the sample
- Allow stain to incubate overnight at RT in a dark room
- To prevent stain from evaporating overnight: Use a large glass petri dish. Soak a paper towel with PBS and place it at the bottom of the petri dish. Use 2 toothpicks to support the slide above the paper towel. This prevents any wicking action from occurring and pulling the stain off the slide.
- Wash 3 times with 1X TBS (submerge in 1X TBS for 5 minutes per wash)
- Place a cover slide over sample. Image sample using fluorescent microscope
