McClean: Registering objectives on the Nikon: Difference between revisions

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[[Image:TI_Setup_Tools_OpenScreen_crop.png]]
[[Image:TI_Setup_Tools_OpenScreen_crop.png]]


*Click ''Setting''
*Click ''Setting''.  A new window will appear.
*Click the appropriate number under ''Nosepiece Data'' (number corresponds to the nosepiece position you want to register the objective at).
*Click the appropriate number under ''Nosepiece Data'' (number corresponds to the nosepiece position you want to register the objective at).
*Select the appropriate ''Series'', ''Mag'' and ''Type'' options in the pull down menus and then select your objective from the list in the selection box below.  For our 100x and 40x objectives you would select:
*Select the appropriate ''Series'', ''Mag'' and ''Type'' options in the pull down menus and then select your objective from the list in the selection box below.  For our 100x and 40x objectives you would select:
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The following images show what the appropriate settings for ou 100x and 40x objective look like for position 3 (100x) and position 4 (40x):
The following images show what the appropriate settings for your 100x and 40x objective look like for position 3 (100x) and position 4 (40x):


==References==
==References==
Adapted from Maitreya Dunham's protocol ([http://dunham.gs.washington.edu/protocols.shtml| Dunham Lab Protocols]) which was adapted from Katja Schwartz's protocol from the Botstein lab ([http://www.princeton.edu/genomics/botstein/protocols/| Botstein Lab Protocols])
[[http://www.nikon.com/products/instruments/support/index.htm Nikon Support]]
 
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==
==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 <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.
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 <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.
*[[User:Megan N McClean|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==
==Contact==
<!--Change the information below to your info if you add a new protocol-->
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*'''[[User:Megan N McClean|Megan N McClean]] 14:01, 03 November 11(EDT)'''
*'''[[User:Megan N McClean|Megan N McClean]] 17:01, 04 July 12(EDT)'''


or instead, [[Talk:{{PAGENAME}}|discuss this protocol]].  
or instead, [[Talk:{{PAGENAME}}|discuss this protocol]].  


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Revision as of 13:48, 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(Ti Setup Tools)" software:
    • Start->All Programs->Nikon->TI Control->TiControl

  • Click Setting. A new window will appear.
  • Click the appropriate number under Nosepiece Data (number corresponds to the nosepiece position you want to register the objective at).
  • Select the appropriate Series, Mag and Type options in the pull down menus and then select your objective from the list in the selection box below. For our 100x and 40x objectives you would select:
    • For the 100x: Plan Apo VC 100x Oil (its the only option that comes up: --- --- PFS 0.13 1.4 1.4)
    • For the 40X: Plan Apo 40x Dry (its the top option that comes up in the box: --- --- PFS 0.14 0.95 0.95)
  • When you pick the appropriate objective in the selection box, and illustration of the objective will show up. This should correspond to the writing on the objective you are trying to register. Check that they match up.
  • Click OK
  • You should get a message saying "Message Sending Succeed"
  • Exit Ti Setup Tools


The following images show what the appropriate settings for your 100x and 40x objective look like for position 3 (100x) and position 4 (40x):

References

[Nikon Support]


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.


Contact

or instead, discuss this protocol. -->