MICROSCOPE: Axio Observer.Z1 SR, inverted, motorized
PALM/STORM RESOLUTION | |
|---|---|
XY | ~ 20 - 30 nm |
Z | ~ 50 - 80 nm |
3D STORM axial range with double phase ramp | ~ 1400 nm |
SIM RESOLUTION | |
|---|---|
XY | ~ 100 - 120 nm |
Z | ~ 200 - 300 nm |
SIM axial range | up to 20 µm into tissue |
IMAGING MODES |
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Photoactivation Localization Microscopy (PALM) |
Strucured Illumination Microscopy (SIM) |
Widefield (WF) (sample illumination with arc lamp) |
Laser Widefield (laser sample illumination in EPI, HiLo or TIRF mode) |
LIGHT SOURCES | ||
|---|---|---|
Light source | Wavelengths [nm] | Nominal power [mW] |
HR Diode | 405 | 50 |
HR Diode | 488 | 200 |
HR DPSS | 561 | 200 |
HR Diode | 642 | 160 |
X-Cite Lumen Dynamics (XLED) | UVX 375 - 400, BDX 450 - 495, GYX 540 - 600 | N/A |
OBJECTIVE LENSES | |||||
|---|---|---|---|---|---|
Objective | Magnification | Immersion Medium | Numerical Aperture | Coverslip correction [mm] | Working Distance [mm] |
EC Plan-Neofluar | 10x | air | 0.3 | 0.17 | 5.2 |
LCI Plan-Neofluar Imm Korr DIC M27 | 25x | water | 0.8 | adjustable, 0 - 0.17 | 0.21 at cover glass 0.17 |
EC Plan-Neofluar OIL DIC M27 | 40x | oil | 1.3 | 0.17 | 0.21 |
C-Apochromat Korr M27 | 63x | water | 1.2 | adjustable, 0.14 - 0.19 | 0.28 at cover glass 0.17 |
Plan-Apochromat DIC M27 | 63x | oil | 1.4 | 0.17 | 0.19 |
alpha Plan-Apochromat DIC M27 Elyra | 100x | oil | 1.46 | 0.17 | 0.11 |
FLUORESCENCE FILTER SETS |
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Two 6-position emission filter turrets are available for imaging on the Zeiss ELYRA PS.1, one with single-bandpass filters and another one with multi-bandpass filters. |
Single-band emission filter set | |||
|---|---|---|---|
Turret position | Emission filter | Excitation filter | Example Fluorophores |
1 | Transmission light | N/A | N/A |
2 | Triple-band filter (labelled None) | N/A | most common fluorophores in the visible range |
3 | BP 420 - 480/LP 750 | MBS 405 | DAPI |
4 | BP 495 - 550/LP 750 | MBS -488 | GFP, Alexa Fluor 488 |
5 | BP 470 - 620/LP 750 | MBS -561 | mCherry, Texas Red |
6 | LP 655 | MBS -642 | Cy5, Alexa Fluor 647 |
| Multi-band emission filter set | |||
|---|---|---|---|
Turret position | Emission filter | Excitation filter | Example Fluorophores |
1 | Empty | N/A | N/A |
2 | Empty | N/A | N/A |
3 | BP 420 - 480/BP 570 - 640/LP 740 | MBS 405/561 | DAPI & mCherry |
4 | BP 420 - 480/BP 495 - 550/LP 650 | MBS 405/488/642 | DAPI, GFP & Cy5 |
5 | LBF -561/642 | MBS -561/642 | Alexa Fluor 555 & Alexa Fluor 647 |
6 | LBF -488/561 | MBS 405ac/488/561 | DAPI, GFP & mCherry |
CAMERA/DETECTOR | |||
|---|---|---|---|
Camera | Pixels | Pixel Size [µm] | Quantum Efficiency (QE) |
Andor iXon 897 (EMCCD) for PALM imaging (left side port) | 512 x 512 | 16 x 16 | 90% |
pco.egde sCMOS for SIM imaging (base port) | 1280 x 1280 | 6.5 x 6.5 | 57%, dynamic range 15 bit |
INCUBATION |
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The microscope is fully enclosed in a blacked out chamber and equipped with a TokaiHit unit (INU series, Model: ELY) |
SOFTWARE |
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ZEN Black Version 11.0.2.190 |
ADDITIONAL INFORMATION |
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Z drive DC servomotor (smallest Z step size 25 nm) |
motorized XY piezo scanning stage motorized (range 130 mm x 100 mm, max. sped 100 mm/s, step resolution 0.2 µm) |
Z-piezo stage insert for XY scanning stage (max. travel range 100 µm, min. 5 nm step size) |
stage inserts available for standard slides, glass-bottom dishes and LabTek chambers |
definite focus |
EPI, HILO and TIRF illumination mode |
image reconstruction algorithms for SIM and PALM data |