TEMVisualizationofMicrotubules

LEVELII

Materials

• CoatedgridforTEM
• 0.1Mammoniumacetate
• 5%ethanolsaturateduranylacetate
• Transmissionelectronmicroscope

Procedure

1. Attheconclusionofa37°CincubationfromExercise9.6,removeonedrop(approximately20microliters)oftheextract,placeitonacollodionorformvarandcarbon-coatedEMgrid,andallowittoremainforabout20-30seconds.6

2. Rinsethegridcarefullywith0.1Mammoniumacetate,andfloatthegridupsidedownonadropof5%uranylacetatefor15minutes.Rinsethegridwithwater,blotallexcessfluidsfromthegrid,andallowittoairdry.

3. Examinethestainedgridinatransmissionelectronmicroscope.

4. Draworphotographyourobservations--calculateallappropriatedimensions(length,diameter)oftheformedtubules.

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