Background

The CSIF houses a Zeiss LSM 510 Confocal Laser Scanning Microscope equipped with a Coherent Mira 900 tunable Ti:Sapphire laser for two-photon excitation. The system was installed late January 2001.

Zeiss provides a free, downloadable LSM IMAGE BROWSER capable of opening and coverting Zeiss .LSM files

Unique features of the LSM 510 system include:

Simultaneous collection of single and multiphoton excitation of different fluorophores

Capability to quickly change the settings for lasers and detectors (a single track) using the digital signal processor (DSP). This allows you to scan across a line with one excitation wavelength and back with a second excitation wavelength. No simultaneous detection, no cross talk at all.

Ability to scan ROIs within a sample - circles, rectangles or freehand regions may be specified. Allows FRAP experiments to be performed with maximum precision.

Ability to perform xz scans along curved lines.

To enable two-photon excitation, we have a tunable Ti:Sapphire laser (700 - 1000 nm). The benefits of two-photon excitation include:

• less bleaching of sample
• analysis deeper into the specimen

For an detailed explanation of how the Ti:Sapphire laser is used, go to the MultiPhoton Excitation Microscopy page at the Coherent web site and download the "MPE tutorial."

System specifications

Microscope - Axiovert 200M inverted, fully motorized microscope

Objectives -

empty	Name/magnification	immersion	NA	working distance	DIC
	Plan-Neo 10X	air	0.30	5.6mm	no
	Fluar 20X	air	0.75	0.66 mm	yes
	Plan-Neo 40X	oil	1.3	0.17 mm	yes
	C-Apochromat 63X	water	1.2	0.24 mm	yes
	Plan-Apo 63X	oil	1.4		yes
	AchroPlan 63X	water	0.9	2.2 mm	no
	Plan-Apo 100X	oil	1.4		yes
	Fluar 100X strain-free	oil	1.3		yes

Light sources - The microscope is equipped with a mercury arc source and filter blocks for DAPI/FITC/Rhodamine for identification of labelled specimens by eye.

The microscope has a transmitted light channel for capturing DIC images in addition to the fluroescent channels. For fluorescent collection, the system has the following lasers with these respective lines:

• LSM 510 W.S software, version 3.2
• LSM Flex Scan package
• Physiology software, v 3.2
• Software 3D for LSM, v 3.2
  

Manual

To obtain a copy of the CSIF user manual for this system, you will need to have Adobe Acrobat reader installed on your computer.

Download a copy of CSIF user manual - LSM 510

Download CSIF server information sheet - explains where data is stored, how long it is stored, and how to ftp from the CSIF computers to your computer.