Confocal and Advanced Light Microscopy (CALM) Facility

Leica TCS SP5 confocal laser scanning microscope

Equipment details and manuals

Confocal laser scanning microscope for multi-dimensional image acquisition and live cell imaging

This system is based on an inverted DMI 6000 CS microscope base, and the scanhead (SP5C) is equipped with three photo multiplier tubes (Hamamatsu R 9624). The system is also equipped with an environment control chamber to maintain environmental conditions for live cell imaging (temperature, 5% carbon dioxide concentration and air humidity). 5 lasers, including a 405 nm blue diode laser, provide 9 different laser lines and thus high flexibility for fluorophore excitation. The system includes LED-based epifluorescence illumination with DAPI, FITC and Rhodamin filter cubes (see below).

 

Lasers

The SP5 provides the following lasers with the specified laser lines for fluorophore excitation:

Laser lines [nm]

Arg

458   

476  

488  

496  

514

HeNe 1

543

     

 

HeNe 2

594

     

 

HeNe3

633

 

 

 

 

Blue diode     

405

 

 

 

 

 

Objectives

The SP5 is equipped with the following lenses:

Lens

magnification  

immersion   

NA

HCX PL APO, 0.17CS

63x

W

1.2

HC PL APO, CS

10x

A

0.4

HC PLAN APO, PH2 0.17/C 0.59     

20x

A

0.7

HCX PL APO PH3 CS

40x

O

1.25

HCX PL APO, lbd. BL

63x

O

1.4

PI APO

100x

O

1.4

W = water, A = air, O = oil

Filter sets

The following filter sets for epifluorescence illumination are available on the SP5:

A UV BP340-380 400 LP 425; I3 blue BP 450-490 510 LP 515; N2.1 green BP 515-560 580 LP 590

For more technical details of the SP5 system, please refer to the technical documentation below.

 

 

Optional:

Manuals for the operation of the Leica SP5

This website provides 3 different manuals that describe how to operate the SP5:

CALM SP5 manual - This is an illustrated manual that explains how to use the system in step-by-step protocols. Click the button to acces the manual:

 

LeicaDMI6000 user manual - This manual by Leica Microsystems gives a detailed description of the DMI6000 inverted microscope base

 

Start-up and run-down procedures

Quick guide for step-by-step protocols for the start-up and closing-down of the SP5 are given at the following link: