Welcome to the Department of Biology

Department of Biology at Illinois State University
Department of Biology

Welcome to the Confocal Microscopy Facility, School of Biological Sciences at Illinois State University

Located in the Science Lab Bldg, College & Fell Av., Normal IL 61790
Supported by funding from the National Science Foundation.
Formal Confocal Training is available through the 2-cr hr course BSC 411; contact K. Edwards for details.
Contact Kevin Edwards for confocal questions or assistance.


Feb. '17: the Ar laser is down for repair. The 561 and 633 lines are still available for use. Contact K. Edwards for more info.

Components and capabilities of the ISU Confocal Facility

Leica TCS SP2 Confocal Microscope (Installed 2004)

Microscope: Inverted Leica DM IRE2 HC Fluo TCS 1-B

- Outfitted for DIC optics and standard fluorescence
-Galvanometer driven z-stage
-Trinocular head and color CCD camera, allowing for capture of nonconfocal images

Objectives

HC PL APO 20x/0.70 Multi-immersion- High numerical aperture Plan Apochromat Immersion lens for Water, Glycerol or Oil Immersion.

HCX PL APO 40x/1.25-0.75 Oil- High numerical aperture Plan Apochromat lens for Oil Immersion.

HCX PL APO 63x/1.4 - 0.6 Oil- High numerical aperture Plan Apochromat lens for Oil Immersion.

HCX PL APO 63x/1.20 W Corr/-CS- High numerical aperture Plan Apochromat Immersion lens for Water Immersion with correction collar. For use without coverglass.

Confocal Scan head: TCS SP 2 Basic Module with spectral detection. Features:

3 Laser sources:
Red (HeNe 633 nm)
Light Green (561 nm DPSS)
Blue (Ar 458-476-488-514 nm)

6 channel Acousto Optical Tunable Filter (AOTF) for laser selection and attenuation.

Three channels for confocal fluorescence or reflected light.

Low-noise photomultipliers, 8 or 12 bit digitization

Transmitted light detector for recording bright field images

Spectral range of detector optics 400-800nm

Adjustable pupil illumination using Beam Expander

Variable, computer controlled detection pinhole diameter

Scan resolution up to 4096 x 4096 pixels

Zoom 1-32x

Scan rotation -2+92 degrees

Scan field diagonal 22mm (intermediate image plane)

LCS Leica Confocal Software incl. 3D, Physiology, & Multicolor packages.

Schedule:

Once you are approved to use the confocal, Dr. Edwards will provide a link to the online reservation calendar (contract him via e-mail).

Resources:

The following faculty can be consulted for advice on fluorescence techniques and imaging:
Edwards: general confocal information; insects; fluorescent protein methods
Gatto: cell culture; biochemistry
Hammond: fungal samples; fluorescent protein methods
Kirik: plants; live imaging
Sedbrook: plants; live imaging
Stein: neural imaging
Vidal-Gadea: C. elegans; neural imaging
Vogel: FACS; mouse tissues; immunological methods

Protocols:

TIPS FOR BETTER CONFOCAL IMAGES: Strategies to fight the battle of signal vs. noise.

Schematic diagram of the scan head:

confocal