This setup combines the ELYRA system (SIM, PALM and TIRF) with a LSM 780 laser scanning confocal microscope. Besides the super-resolution techniques for which ELYRA is primarily designed, ELYRA’s high end cameras can be used for widefield imaging applications requiring high sensitivity of detection such as single molecule tracking. The motorised stage allows multi-position time-lapse imaging or tile scans of large samples. A heated stage top chamber is available upon request.

Available techniques:

• Structured illumination microscopy (SIM) in 3D
• Single molecule localization microscopy (PALM, STORM, PAINT, …)
• Single particle tracking (SPT) and imaging fluorescence correlation spectroscopy (ImFCS)
• Total internal reflection fluorescence (TIRF) microscopy and Hi-Lo mode imaging
• Laser scanning microscopy (LSM) confocal imaging
• Fast spectral imaging and linear spectral unmixing
• Fluorescence recovery after photobleaching (FRAP)

Objectives:

• EC Plan-Neofluar 10x/0.3 dry, FWD 5.2 mm, CG 0.17 mm
• Plan-Apochromat 20x/0.8 dry, FWD 0.55 mm, CG 0.17 mm
• EC Plan-Neofluar 40x/1.3 oil, FWD 0.21 mm, CG 0.17 mm
• LD Plan-Neofluar 63x/0.75 dry, FWD 1.2-2.2 mm, CG 0-1.5 mm
• Plan-Apochromat 63x/1.4 oil, FWD 0.19 mm, CG 0.17 mm
• Alpha Plan-Apochromat 100x/1.46 oil, FWD 0.11 mm, CG 0.17 mm
  [FWD = free working distance, CG = cover glass]

Fluorescence excitation sources:

ELYRA laser lines (intensities at sample):

• 405 nm, 488 nm, 561 nm, 642 nm

LSM 780 laser lines (power at sample):

• 405 nm, 458 nm, 488 nm, 514 nm, 561 nm, 633 nm

Epifluorescence:

• X-Cite mercury metal halide lamp

Filter sets and dichroics:

ELYRA filter sets:

• Zeiss 77 HE (483+564+642; 506+582+659; 526+601+688)
• BP 420-480 + LP 750 emission filter
• BP 495-575 + LP 750 emission filter
• BP 570-650 + LP 750 emission filter
• LP 633 emission filter

LSM 780 dichroics:

• 405 nm; 445 nm; 458 nm; 458/514 nm; 458/561 nm; 488 nm; 488/561 nm; 488/561/633 nm; T80/R20 partially reflective mirror

Detectors and cameras:

ELYRA cameras:

• Andor iXon DU-897D, 512x512 pixels, 16 µm/pixel
• pco.edge sCMOS, 1280x1280 pixels, 6.5 µm/pixel

LSM 780 detectors:

• 32-GaAsP spectral array (410 - 695 nm)
• 2 PMTs
• Transmitted light PMT

Software:

• ZEN 2012 SP5 FP2 (black)

Other features:

• Motorised stage
• Motorised filter cube turret
• Motorised focus drive
• Stage-top incubator (on request)

Usage fees* [SGD/hour]	NTU	Academia	Industry
Confocal, TIRF	30	45	72
SIM, Localisation microscopy		60	96
Location	ABIF, SCELSE, B2 (SBS-B2n-27P.2)
Safety Notice	Requires N3 laser license (N2/04107/0169)
Contact	abif@e.ntu.edu.sg; nobic.facilities@e.ntu.edu.sg

* Long-term discounts of 75% and 95% apply after 8 and 24 hours of continuous usage, respectively.

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