McClean: Registering objectives on the Nikon: Difference between revisions
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==Protocol== | ==Protocol== | ||
To register an objective or change the registration settings of an objective: | |||
*Open the "TI-Control" software: | |||
* | **Start->All Programs->Nikon->TI Control->TiControl | ||
* | [[Image:TI_Setup_Tools_OpenScreen_crop.png]] | ||
* | |||
==References== | ==References== |
Revision as of 13:39, 4 July 2012
Overview
To use the PFS (perfect focus system) motorized nosepiece the objective information needs to be correctly registered on the microscope. Currently the objectives that we use most often (100x Plan Apo VC oil; 40x Plan Apo dry) are correctly registered. The 40x is registered to position 4 (and 1) and the 100x is registered to position 3 (and 2,5,6).
If you add a new objective that is compatible with the PFS, you will need to follow the procedure below to register it.
If the PFS is not working, it may be that the objectives are not registered correctly. Check the front display of the microscope to make sure that it displays information on the objective you are using (PA40x or PV100x). If it does not it is likely that the objectives are not registered or are not registered to the position you are using on the objective turret. To fix this, follow the directions below to register the objective you want to use to the appropriate turret position.
Protocol
To register an objective or change the registration settings of an objective:
- Open the "TI-Control" software:
- Start->All Programs->Nikon->TI Control->TiControl
References
Adapted from Maitreya Dunham's protocol (Dunham Lab Protocols) which was adapted from Katja Schwartz's protocol from the Botstein lab (Botstein Lab Protocols)
The strains were made in the Thorner lab. See Julius et al (1983) Cell, 32, 839-52 for documentation of DBY7730. DBY7442's exact genotype is unknown. The sst mutations make them super-sensitive to pheromone. When exposed to pheromone of the opposite mating type, the cells arrest.
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input! Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
- Megan N McClean 11:02, 3 November 2011 (EDT) We have been having some trouble lately with the yMM422 strain. Strains of the opposite mating type have not been forming scorable halos. We may want to try fresh stock from the Botstein lab's -80°C stock. Alternatively, it seems to work 'sometimes' and since none of us are that careful about how many cells we seed or what growth phase they are in when we do so, perhaps we should play around with that and then be consistent in the future.
Contact
- Megan N McClean 14:01, 03 November 11(EDT)
or instead, discuss this protocol.