leica-sp5-resonant_s

Location: Room 53

System specifications

Leica SP5 inverted confocal microscope with aresonant scanner allowing high-speed confocal imaging and is well suited for live-cell imaging and high-resolution morphological imaging.

350-900 nm spectral detection range with 5 nm spectral resolution and sub-micron spatial resolution

  • 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 and live specimen (Enviromental chamber).

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 (additional)

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

Data structure: Setting record (*.lei) and 8-bit images (*.tif)

Image size: 64 x 64-8000 x 8000 pixels

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, PMT4, PMT5
  • 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

 

561

RedFPs, Alexa568

 

594

Alexa 594, mCherry

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

20x *

0.70

PL Apo UV

Imm/Corr

- / 0.17

40x

1.25

PL Apo UV

Oil

0.17

63x

1.20

PL Apo UV

Water

0.14 - 0.18

63x

1.4

PL Apo UV

Oil

0.17

* The 20x immersion objective for water, glycerine and immersion oil

Fluorescent source

Leica EL6000 External light source for fluorescence excitation (alignment-free, mercury metal halide bulb with an extremely long lifetime of more than 3000 hours)

Fluorescence filter sets

Name

Excitation filter

Emission filter

Dichroic

A

BP 340 - 380 nm

LP 430 nm

400 nm

CFP

BP 436/20 nm

BP 480/40 nm

455 nm

GFP

BP 470 40

BP 525/50 nm

580 nm

N2.1

BP 515 - 560 nm

LP 590 nm

580 nm

YFP

BP 500/20 nm

BP 535/30  nm

515 nm

Power up and down procedure

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

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