Andor DragonFly (room 3103)

This is a microscope set up for live imaging with a fully motorized xy stage for multi-position, tile scan and z Piezo for fast 3D stacks.

Imaging platform

Photo: UiO

Information about the microscope

This system is a set up for live cell imaging and equipped with heating, humidified incubation chamber, 5% CO2 and Nikon perfect focus system (hardware autofocus).

DragonFly allows the use of various imaging modalities. Transmission and fluorescence in wide field and spinning disk confocal modes. For the spinning disk confocal, two pinholes can be selected: a small 25 µm pinhole for best z sectioning and a larger 40 µm pinhole for best signal to noise ratio. DragonFly can also be used for TIRF imaging. A GPU-accelerated deconvolution function is integrated.

Basic principle of spinning disk confocal microscopy:

 

The DragonFly is equipped with two cameras:

  • iXon EM-CCD 1024x1024 has the highest sensitivity (95% peak QE, up to 53 full frames per second)
  • Zyla4.2 sCMOS 2048x2048 the highest resolution and speed (80% peak QE, up to 400 fps in spinning disk mode, 1000 fps in wide-field and TIRF)

Each camera is equipped with zoom optics for 1x/1.5x/2x additional zoom factor. The two cameras can be used individually or together (dual camera) for simultaneous acquisition of two color channels.

The combination of powerful lasers and sensitive camera also makes it possible to use the system for single molecule localization microscopy (dSTORM) to achieve super-resolution (30 nm lateral resolution). A cylindrical lens is installed to achieve super-resolution also in 3D. Wide-field, spinning disk and TIRF modes can also be used to achieve super-resolution by SRRF (50-150 nm lateral resolution, no improvement in z, ~1 fps for 512x512, processing in imageJ) that does not need special fluorophores or sample preparation.

Additionally, DragonFly is equipped with a FRAPPA unit (micro mirror array) for photobleaching/photoactivation of freely definable regions using a high power 405 nm diode laser (1150 mW).

For more information on DragonFly see the maufacturer's website (oxinst.com)

Imaging laser lines:

Wavelength

Type

Power output*

405 nm

diode

100 mW

488 nm

diode

150 mW

561 nm

diode

100 mW

637 nm

diode

 140 mW

* from fiber

Filter sets:

Filter/dichroic

 

comment

main dichroic (quad)

 

                405/488/561/640

 

DAPI

                       450/50BP

 

GFP

                       525/50BP

 

RFP

                       600/50BP

 

Cy5

                       700/75BP

 

beam splitter

                           570LP

dual camera mode

eyepieces: DAPI/GFP/TxRed

Objectives***

Magnification

Type

Numerical aperture

Working distance (µm)*

Field of view width (µm)**

Immersion

comment

4x

CFI Plan Apo

0.2

20000

3090

-

 

10x

CFI Plan Apo

0.45

4000

1235

-

 

20x

CFI Plan Apo

0.75

200

617

-

 

40x

CPI Plan Fluor

1.3

200

309

oil

 

60x

CF Apo

1.4

140

206

oil

 

100x

CFI Plan Apo

1.45

130

123

oil

TIRF

60x

CFI Plan Apo

1.2

270

206

water

 

40x

CFI Plan Apo

0.75

660

309

-

 

* from coverslip #1.5 (170 µm)

** effective camera field of view at 1x camera zoom

*** 6 installed in turret, optional (on request) as of September 2018: 4x dry and 40x dry.

Published Dec. 1, 2020 2:36 PM - Last modified Mar. 2, 2021 5:45 PM