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LOCATION: Room 09.171.1 (BSL2)

SYSTEM SPECIFICATIONS:

  • Imaging: Fast imaging with low phototoxicity
  • Stand: Nikon Ti2 with perfect focus system (PFS)
  • XY-stage: motorized
  • Temperature and CO2 incubation
  • Software: NIS elements with JOBS
  • Yokogawa CSU-W1 confocal scan head (50um disk)

CAMERA SPECIFICATIONS:

  • Model: 2x Hamamatsu ORCA-Fusion, sCMOS
  • Effective number of pixels: 2304 x 2304
  • Cell size: 6.5µm x 6.5µm
  • Effective Area: 14.976mm x 14.976mm
  • Max frame rate: 100 frames/second
  • Dynamic range: 16 bit
  • Quantum efficiency (peak): > 80% (@550nm)
  • Read out noise: 0.7 - 1.4 electrons (depending on fps)

OBJECTIVES

Objective

Magnification / NA

Medium

Contrast

Working Distance

CFI Plan Apo Lambda D

4x / 0.2

air

BF

20 mm

CFI Plan Apo Lambda D

10x / 0.45

air

BF

4 mm

CFI Plan Apo Lambda S

25x / 1.05

silicon

BF

0.55 mm

CFI Apo LWD Lambda S

40x / 1.15

water

BF

0.6 mm

CFI Plan Apo Lambda D 60x / 1.42 oil BF 0.15 mm

CFI Plan Apo Lambda 

100x / 1.45

oil

BF

0.13 mm

ILLUMINATION lasers for spinning disk

Wavelengths (nm)

Type

Some dyes and use

405

Diode laser

Alexa405, DAPI

488

Diode laser

Alexa488, GFP

561

Diode laser

Alexa568, RFP

640

Diode laser

Alexa647, Cy5

ILLUMINATION LED for widefield and eyepiece observation

Wavelengths (nm)

Type

Some dyes and use

385

LED

Alexa405, DAPI

475

LED

Alexa488, GFP

550

LED

Alexa568, RFP

621

LED

Alexa647, Cy5

Main DICHROIC MIRROR

Fluorochromes

Transmission to sample (nm)

Reflection to camera (nm)

DAPI

> 80% 405 – 405 nm

> 95% 422 – 473 nm

GFP, Alexa488

> 80% 488 – 488 nm

> 95% 503 – 545 nm

RFP, Alexa568

> 80% 620 – 665 nm

> 95% 586 – 620 nm

Cy5, Alexa647

> 80% 638 – 647 nm

> 95% 665 – 750 nm

Secondary DICHROIC MIRROR

Single camera mode

Full mirror to primary camera (back port)

Dual camera mode

LP 561 nm beam splitter

EMISSION FILTERS in front of the primary camera (back port)

Fluorochromes

Availability

Bandpass

Transmission (nm)

DAPI

YES

single BP

417-477

GFP, Alexa488

YES

single BP

510-540

RFP, Alexa568

YES

single BP

574-626

Cy5, Alexa647

YES

single BP

671-745

EMISSION FILTERS in front of the secondary camera (left port)

Fluorochromes

Availability

Bandpass

Transmission (nm)

RFP, Alexa568

YES

single BP

574-626

Cy5, Alexa647

YES

single BP

671-745

Filter cubes (sets) for eyepiece observation

Fluorochromes

Excitation filer (nm)

Dichroic cutoff (nm)

Emission filter (nm)

DAPI

377/50

409

447/60

GFP, Alexa488

466/40

495

525/50

RFP, Alexa568

562/40

593

641/75