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Nikon Ti2 X-Light V3

LOCATION: Room 07.057

SYSTEM SPECIFICATIONS:

  • Imaging: Fast imaging / tiling of full tissue sections not thicker than 50 um
  • Confocal and widefield mode
  • Confocal scanhead: X-light Crest v3
  • Disk: 50 um pinhole size, 250 um spacing
  • Stand: Nikon Ti2 with perfect focus system (PFS)
  • XY-stage: motorized
  • Software: NIS elements with JOBS
  • Lightsource: Lumencor Celesta 7 channel including CFP/YFP and Cy7/A750

CAMERA SPECIFICATIONS:

  • Model: Teledyne Photometrics Kinetix
  • Mount: 25 mm, large field of view
  • Effective number of pixels: 3200 x 3200 cropped to 2700 x 2700
  • Pixel size: 6.5µm x 6.5µm
  • Effective Area: 14.976mm x 14.976mm
  • Max frame rate: 88 frames/second
  • Dynamic range: 12 bit
  • Read out noise: 1.2 electrons

OBJECTIVES

Objective

Magnification / NA

Medium

Contrast

Working Distance

CFI Plan Apochromat
Lambda D

4x / 0.2

air

-

20 mm

CFP Plan Apo Lambda D

10x / 0.45

air

-

4 mm

CFI Plan Apo

20x / 0.75

air

-

1 mm

CFI Plan Apo Lambda S

25x / 1.05

silicon

-

0.55 mm

CFI Plan Apo Lambda

60x / 1.4

oil

-

0.13 mm

ILLUMINATION Laser excitation (Lumencore Celesta)

Wavelengths (+- 2 nm)

Some dyes and use

405

Alexa405, DAPI

446

CFP

477

Alexa488, GFP

520

YFP

546

Alexa555, 568, CY3

638

Alexa647, CY5

742

Alexa750, CY7

EXCITATION FILTERS in the X-Light v3 scanhead

Filter set (multi-band)

excitation (nm)

V3-DA/FI/TR/Cy5/Cy7 391/477/549/639/741
V3-CFP/YFP 449/520
5% transmission ND for widefield mode only
DICHROIC MIRRORS in the X-Light v3 scanhead
Filter set (multi-band) dichroic mirror (nm)

V3-DA/FI/TR/Cy5/Cy7

441/511/593/684/817
V3-CFP/YFP 471/539

EMISSION FILTERS in front of the camera

Fluorochromes

Bandpass

Transmission (nm)

DAPI

single BP

438/24
CFP single BP

485/20

GFP, Alexa488

single BP

511/20
YFP single BP 560/25

CY3, Alexa555, 568

single BP

595/31

CY5, Alexa647

single BP

685/40
CY7, Alexa750 multiband 421/511/593/684/817