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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

A


 

SIM RESOLUTION 

XY

~ 100 - 120 nm 

Z

~ 200 - 300 nm

SIM axial range

up to 20 µm into tissue

A


IMAGING MODES

  • Photoactivation Localization Microscopy (PALM)
  • Structured 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

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

A


FLUORESCENCE FILTER SETS

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

A


 

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

A


 

INCUBATION

The microscope is fully enclosed in a blacked out chamber and equipped with a TokaiHit unit (INU series, Model: ELY)

SOFTWARE

ZEN Black Version 11.0.2.190

ADDITIONAL INFORMATION

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

 

 

 

 

 



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