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  1. Plan ApoN 60x/1.42

  2. TIRF Apo 60x/1.49

Cameras and Emission Filters
  • Camera 1: DAPI: 435/31 Cy5: 683/40 DIC

  • Camera 2: A488: 528/48

  • Camera 3: A568: 609/37

  • 405 nm

  • 488 nm

  • 568 nm

  • 640 nm

Sample holder compatibility
  • Standard microscope slides

  • 35 mm glass bottom dishes

  • Multiwell chambered coverslips

Interface Requirements

  • Linux Centos 6.8

  • Kernel Linux 2.6.32-642.13.1.e16x86_64

  • GNOME 2.28.2

  • Intel Core i7 2600 CPU
  • NVidia NVS300
  • Drives 1 TB SATA boot drive
    RAID 3 × 1 TB SATA in RAID5 configuration

Notes on usage:

SIM imaging is extremely sensitive to spherical aberration. To get the highest quality images you need to minimize spherical aberration. This means:

  • You should use high precision 0.17 mm coverslips

  • You should mount only a single coverslip in the center of the slide.

  • You should mount in an appropriate mounting medium. Fixed samples should be mounted in high refractive index mounting media with antifade. Be aware that hard-set mounting medium can cause shrinkage when it cures. We also recommend that you do not use mounting medium which contains DAPI or other fluorophores.

  • You should have your sample as close to the coverslip as possible (e.g. cells grown on coverslip not on slide).

  • You should use bright and stable fluorophores.

  • You should select the correct immersion oil for you sample. You can use the GE immersion oil calculator to determine the best oil to start with. The also have the calculator as a phone app.

As a general rule, if the sample doesn't look good in widefield or confocal, SIM imaging will not improve it. You should make every attempt to optimize your staining and reduce background from out-of-focus fluorescence for the best results.

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