sp5-confocal_s

Location: Room 42

System specifications

Confocal microscope system with a broad range of scan speeds suited for high-resolution morphological imaging, as well as multispectral imaging.

  • 350-900 nm spectral detection range with 5 nm spectral resolution and sub-micron spatial resolution
  • Suitable for most fluorophores from DAPI to Cy5
  • Measuring up to five dimensions, including x, y, z, time and spectrum
  • Capable of multi-dimensional fluorescence qualitative and quantitative measurement, FRET, FRAP, FLIP, FLAP and other photobleaching-related applications
  • Suitable for imaging fixed specimen (No environmental chamber).

Internal Detectors

  • Spectral detection channels: PMT1, PMT2, PMT3
  • Brightfield transmission channel: PMT Trans

Multi-dimension capability: xyz, xzy, xt, xyt, xzt, xyzt, xy, xz, xyt, xyz

Stage

  • Manual moving X, Y stage
  • Motorized z stage (piezo-controlled) with 166.68 µm scanning range

Zoom range: 1x-32x

Pinhole size: 20 µm-600 µm

Contrast enhancement options: Brightfield, DIC and polarized

Software: Leica LAS AF, Version 1.7.0, for image acquisition and processing

Scanners

Conventional scanner: 10-1400 Hz (2-3 frames/sec at 512 x 512 pixels)

Internal detectors

Reflection/fluorescence mode (spectral detection range 400-800 nm)

  • Spectral detection channels: PMT1, PMT2, PMT3
  • Additional PMT in transmission mode

Lasers

Type

Wavelenght (nm)

Dyes and use

UV

405

DAPI, Hoechst

Argon

458, 476, 488, 496, 514

CyanFP, GFP, Alexa488, CY2, YFP

HeNe

543

Alexa 546, Alexa 555

HeNe

633

Cy5, Alexa 647

  • An acousto-optical tunable filter (AOTF) controls the intensity of the visible excitation laser light.
  • An acousto-optical beam splitter (AOBS) is used to split excitation light and emission light instead of filter blocks.

Microscope

Leica DMI 6000 CS AFC Bino inverted microscope with motorized XY-stage

Objectives

Magnification

NA

        Type

Immersion

Coverslip

WD (mm)

   20x

0.5

PL Fluotar UV optimised

Dry

- / 0.17

0.68

    40x

1.25

HC PL Apo UV optimised

Oil

0.17

0.65

   63x

1.4

HC PL Apo UV optimised

Oil

0.17

0.14

Fluorescent source

Leica EL6000External light source for fluorescence excitation (alignment-free, mercury metal halide bulb with an extremely long lifetime of more than 2000 hours)

Fluorescence filter sets

Name

Excitation filter

Emission filter

Dichroic

A

BP 340 - 380 nm

LP 425 nm

400 nm

I3

BP 450- 490 nm

LP 515 nm

510 nm

N2.1

BP 515-560

LP 590

580 nm

Power up and down procedure

  • Please contact Alvaro Crevenna to schedule a session
  • Report problems on the Log file

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