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Leica CW-STED Super-Resolution Demo

In the middle of July 2010 (July 8-July 21; schedule below), the Microscope Facility is pleased to host a demo of Leica's new CW-STED for two-color fluorescence. It's the first super-resolution technique that balances high-speed (5-25 fps) with high resolution (70nm). Normal confocal microscopy has only ~250nm resolution, so CW-STED is ~4x better!

Schedule for Super-resolution Demo

Immediately:

Immediately: (list of already responded)

Soon: Prep sample (see summary table below)
New Latest details for 2-color sample prep
New Latest details for 1-color sample prep

June 30, 2010, noon, 1830 Bldg Suite 2-200, Bodian Room:
Workshop with Myriam Gastard (Leica) with more details on specimen preparation techniques
Scot's Notes, Slides from Presentation

July 12, 2010, noon:, 1830 Bldg Suite 2-200, Bodian Room:
Joerg Bewersdorf, Asst Prof, Cell Biology, Yale University: Present his research using TiSa-STED, 4Pi and PALM super-resolution techniques to study chromatin structure.

July 12-21, 2010, PCTB G18/19:
Hands-on demo: 28 blocks, 2-3 hr each, 9a-5p
Only applicants (see #2) have access to sign-up site!

If you are interested in testing this new technology (note restrictions on sample) and help us write a shared instrumentation grant:

  1. Immediately: to express interest (if already sent an email, see list at bottom to confirm)
  2. Immediately: (see list at bottom to see if your application was received) with 1-3 paragraph description of:
    • How super-resolution would help your research
    • How you’d get appropriate data (for Shared Instrum Grant) from limited 2hr slot
    Examples:
    Morphology: crowded structures (<200nm) not resolvable with std confocal (side-by-side confocal vs CW-STED)
    Dynamics: subcellular motions of small/crowded structures (i.e. not visible in std confocal because of speed or resolution; comparison movies?)
    Colocalization: regulatable multiprotein complexes?  (side-by-side confocal vs CW-STED)
  3. Soon: prepare samples for demo:
    New Latest details, trade-offs, etc for 2-color sample prep
    New Latest details, trade-offs, etc for 1-color sample prep
     BestGoodAVOID
    Fluor Protein

    mCerulean

    mCitrin

    mVenus

    eCFP

    eYFP (good for 2-color)

    eGFP (variable results)

     
    Dyes

    Dylight488 (best for 2-color)

    Chromeo488

    Alexa 488

    Atto 488

    Oregon Green

    FITC

    DAPI

    Q-Dots

    2nd color dye

    Alexa430
    (other color: eYFP OK, but Dylight488 best)

      
    Mounting Media

    Thiodiethanol (see PDFs) + Antifade

    ProLong (avoid version that contains DAPI) + Antifade

    Moviol + Antifade

    Slowfade

    Vectashield

 

Example of CW-STED

Vimentin
Vimentin-YFP/Keratinocytes (Reiner Windoffer RWTH Aachen Univ.)

 

Already responded (2010 Jun 29)
PIAdd'l contactsApplication Rec'd
(granted access
to sign-up site)
Andrew, Deborah J. Rec'd 6/29/10
Bhujwalla, Zaver
Brown, Amanda Rec'd 6/20/10
Bulte, JefLiang, Yajie
Chen, Elizabeth
Donowitz, MarkChen, Tian-E Rec'd 6/28/10
Edidin, Michael
Ewald, Andrew J.
Fuchs, PaulWersinger, Eric Rec'd 6/10/10
Gabelli, Sandra Beatriz
Gibson, Wade
Griffin, Jack
Hanes, Justin SWoodworth, Graeme
Hoiczyk, Egbert
Hubbard, Ann
Huganir, Richard L.
Kolodkin, Alex
Kovbasnjuk, Olga N Rec'd 6/7/10
Li, Xingde
Meffert, MollieRuiz, Claudia pass
Montell, Denise
Pathak, Arvind
Reddy, Karen Rec'd 6/18/10
Robinson, Douglas
Roden, RichardKwak, Kihyuck
Ross, Christopher A.
Schneck, JonathanBieler, Joan
Sesaki, Hiromi
Sollner-Webb, BarbaraHeydarian, Mohammad Rec'd 6/22/10
Winslow, Raimond L. pass
Worley, Paul
Xiao, Jie

 


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