Equipment and protocols

Spinning Disk 2

Spinning disk confocal microscope with FLIM detection, laser scanning photomanipulation, and millisecond spherical abberation detection.

Configured by Intelligent Imaging Innovations, this microscope is our top instrument for imaging of molecular complexes in living cells.

Laser launch

405 Diode
443 Diode
491 DPSS
515 DPSS
561 DPSS

Crystal Technology AOTF
Galvometer port switcher for confocal, FRAP or FLIM laser launch

Objectives:

63x Plan Apo 1.4 na oil
63x Plan Apo 1.3 na glycerin 21°C
63x Plan Apo 1.2 na water
20x Plan Apo 0.7 na imm

Camera:

Photometrics Evolve 512x512 back-thinned EMCCD

Stage

ASI motorized stage with MCL piezo Z

Confocal head:

Yokogawa CSU-X

Filters

Dichroics

Semrock 405/488/568
Semrock 443/514

Emission

Chroma ET 480/40
Chroma ET 525/50
Chroma ET 560/55
Chroma ET 595/50

FLIM

Frequency domain fluorescence lifetime imaging

Pockel's cell excitation modulation
Lambert gated intensifier for emmision modulation
Photometrics CoolSnap CCD camera

FRAP

Dual galvometer scanner with galvometer path switcher for bleaching and activation of arbitrary regions and rapid switching between photomanipulation and imaging.

mSAC

A movable lens in the imaging path allows for spherical abberation correction. Part of the path switcher, the lens is positioned by a voice coil for rapid positioning so that correction can be performed throughout a rapid z-series.

 

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