Zeiss Elyra PS.1 – University of Copenhagen

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Comparison of SIM (left) and Widefield (right) images of the same structure.


Zeiss Elyra PS.1 Super Resolution Microscope

Elyra is a super resolution microscope with Structured Illumination Microscopy (SIM), and Single Molecule Localization Microscopy (PALM or dSTORM for example).

Structured Illumination Microscopy (SIM) relies on light interfences creating moire fringes and improves the resolution by a factor of 2 (xy: 100nm; z: 250nm). Here are some recommendations for sample preparation for SIM.

Localization microscopy techniques such as dSTORM and PALM take advantage of the blinking properties of some fluorophores to calculate their precise location in a sample. The resolution is greatly improved (xy: 40nm ; z: 100nm via TIRF).

  Imaging Widefield / SIM / PALM / dSTORM


PALM/dSTORM: cooled EMCCD Andor - iXon DU897



Zeiss AxioObserver

Stage Motorized in XYZ
Software Zeiss Zen Black 2012

Objective Mag. / NA Medium Contrast Working Distance
EC Plan Neofluar 10x / 0.3 Air DIC 5.2 mm
Plan-Apochromat 20x / 0.8 Air DIC 0.55 mm
C-Apochromat 63x / 1.2 Water DIC 0.28 at 0.17
Plan-Apochromat 63x / 1.4 Oil DIC 0.19
alpha Plan-Apochromat 100x /1.46 Oil DIC 0.11
alpha Plan-Apochromat 100x / 1.57 Oil-HI DIC 0.11

Lasers Type and Power Some dyes and use
405 HR Diode - 50mW DAPI, Hoechst, AlexaFluor405...
488 HR Diode -200mW GFP, AlexaFluor488...
561 HR Diode -200mW RFPs, AlexaFluor555, 568...
642 HR Diode -150mW AlexaFluor647, DRACQ5, Cy5...

Filter sets for Super Resolution
Set MBS Emission Fluorophores
 3 405 BP420-480/LP750 Blue
488 BP495-575/LP750 Green
561 BP570-650/LP750 Red
642 LP 655 Far Red
2 405/561 BP420-480/BP570-640/LP740 Blue / Red / Far Red
405/488/642 BP420-480/BP495-560/LP650 Blue / Green / Far Red
561/642 LBF561/642 Red / Far Red
405/488/561 LBF488/561 Green / Far Red

FILTER CUBES for Arc lamp (X-cite)
Fluorophores Excitation Dichroic Emission Filter No
Green/Red/Far Red TBP 483+564+642 TFT 506+582+659 TBP 526+601+688 77HE

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