Confocal microscopy


Located in the Science Lab Bldg, College & Fell Av., Normal IL 61790

Supported by funding from the National Science Foundation.

Contact Kevin Edwards for confocal questions or assistance.

ISU is pleased to announce the funding of our new Confocal Microscopy Core Facility. It is funded by a grant from the National Science Foundation. See the award page for more details

We are working to develop this facility through the Fall of 2018 and hope to be training users starting in Jan. or Feb. 2019. Students: We plan to offer the confocal training course BSC411 (2 cr hr) in Spring 2019 semester.

The existing SP2 facility is functional, and if your experiment cannot wait til January, please contact Kevin Edwards [kaedwar@ilstu.edu] for user information.

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; Drosophila; arthropods; fluorescent protein methods
  • Gatto: cell culture; biochemistry
  • Hammond: fungal samples; fluorescent protein methods
  • Kirik: plants; live imaging
  • A. Mortimer: Drosophila
  • 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

Controlled environmental facilities for living organisms


For research with microbes and algae to plants and animals we have a range of facilities for experimental research where controlling the enironmental conditions is key.

Aquarium Room

For fish and shellfish

Includes a 4,800 liter Fresh and Saltwater tank system for sea creatures

Incubators

For bateria, fungi, and flies

EGC Growth Chambers

For algae and plants

Research Greenhouses

Our school has professional staffed greenhouses for plants

Vertebrate Animal Care

ISU maintains a staffed, centralized vivarium for housing many vertebrate species used in research and teaching.

Field ecology and environmental analysis


We use diversity of natural areas and field stations throughout Illinois and neighboring states for observational and experimental studies. Representative sites are listed below. Some of the field and laboratory equipment is listed.

Representative Research Sites

Natural marsh along the Illinois river used for research on the physiological ecology of turtles

Mackinaw Study Area

Forest area in north-central Illinois and site of long-term research on the ecology and behavior of house wrens (Troglodytes aedon). For more information, click here.

Native Prairie Remnants

Many prairie preserves representing the history diversity of prairie types are used for observational studies on effects of habitat fragmentation and deer impacts on communities

Many of these remnants are own and managed by the IL DNR.

Visit their website for more information.

ParkLand Preserves

Mostly forest areas where research has focused on ecology of container-dwelling mosquitos, house wrens, and invasive plants

For more about ParkLands preserves visit their website.

Reconstructed Prairies

Replanted prairies used for more experimental and longer-term studies where some have focused on the ecology of parasitic plants.

Tyson Research Center

Located in Eureka, MO, west of St. Louis, this is the field station of Washington University, St Louis. It is used for experimental studies of ecology of container-dwelling mosquitoes

For more about the Tyson Research Center visit their website

Representative Equipment

Digital Video Equipment

For recording of animal behavior in field and lab studies

A Diversity of Environmental Sensors

For electronic recording temperature, light and other measurements

Li-Cor Li-6400 Portable Photosynthesis System

Lachat Flow Injection Analysis System and Ion Chromatograph

For chemical analysis of soils and water

Ashing and drying ovens

Greenhouse


The Biological Sciences Greenhouse provides a place for plants to grow. There’s lots of sunlight.

Equipment Summary:

  • Sunlight
  • Water
  • Seeds
  • Various temperature control devices

greenhouse

Historic collections


Our school has preserved collections of insects, birds and plants. These on-going collections are a valuable resource for assessment of the impacts of environmental changes.

John Wesley Powel and Dale Birkenholz Natural History Collections

Collections from 1867 of vertebrates including: fish, herps, birds and mamals. For more information see Dr. Capparella’s website who is the curator of this collection.

Entomological Research Collection

A diversity collection of invertebrates

George S. Vasey ISU Herbarium

Collection of over 50,000 specimens of plants dating back to 1859. For more information see Dr. Armstrong’s website who is the curator of this collection

Microscopy and image analysis


Our school has extensive microscopy facilities and training for graduate students.

The Biological Sciences Microscopy Facility provides a range of instruments for light and electron microscopy, including a new, state-of-the-art Scanning Electron Microscope. For usage information, please contact Dr. Martha Cook.

Here you can view the raw data and full TIF figures for the publication,”Rapid phenotypic analysis of uncoated Drosophila samples with low-vacuum scanning electron microscopy” by Tardi, Cook, and Edwards (2012).

Here is a brief summary of the equipment in the Facility:

  • Scanning Electron Microscope
    • FEI Quanta 450 tungsten environmental scanning electron microscope with Oxford Inca 250 EDS. Funding by the National Science Foundation.
  • Transmission Electron Microscope
    • Zeiss EM10
  • Collection of Light Microscopes
    • Including Leica DMRBE equipped with darkfield, DIC, and phase contrast as well as fluorescence filter sets for UV, blue, green and GFP excitation
  • Confocal Microscope Funding by the National Science Foundation.
  • Pelco CPD2 critical point dryer
  • Hummer 6.2 sputter coater
  • Denton DV-502 vacuum evaporator
  • Pelco 3450 microwave oven with temperature probe and load cooler
  • Leica Ultracut UCT ultramicrotome

Many Faculty Research Labs have other microscopes for their particular uses, including:

Zeiss Axiovert inverted microscope w/ Narishige microinjector, needle puller and grinder

fly

More Scope Information

Molecular and cellular biology


Here are some examples of the equipment available for use by our faculty and their students for techniques in molecular and cellular biology.

  • Phosphorimager: Quantifiable digital imaging of radioactive samples
  • ABI DNA sequencer
  • AB 7300 Real-time PCR
  • DNA microarray scanner
  • Steris Amsco Autoclave, Dishwasher and Drier
  • PTI QuantaMaster Fluorometer
  • Techne Hybridization Oven/Incubator HB-1D
  • Typhoon FLA7000IP Modular biomolecular imager
  • GenePix Professional 4200A microarray scanner
  • Thermo French Press
  • Ultracentrifuges and centrifuges including:
    • Beckman Avanti J-25I and J-26S centrifuges
    • Beckman XL-90 ultracentrifuge
    • Beckman TL-100 tabletop ultracentrifuge

  • BD FACSCalibur flow cytometer
    • cell sorting
  • HPLC
  • Tissue culture hoods
  • CO2 incubators
  • Beckman LS 6500 Scintillation counter
  • Biotek Elx800Uv microplate readers
  • Eppendorf Mastercycler Thermocycler
  • Thermo NanoDrop 2000C Spectrophotometer
  • Thermo Fluoroskan Ascent FL
  • UVP Epi Chem Darkroom with MegaCam 810 CCD camera
  • UVP white/UV transilluminator with CCD camera
  • UVP EC3 Imaging System
  • Research / Instruction Cold Room
  • VersaMax microplate reader
  • Taylor-Wharton K series Cyrostorage System
  • ABI 3130 Genetic Analyzer

Neuroscience and physiology


Here are some of the key equipment for use by our faculty and their students in neuroscience.

  • Fluorescence microscope with high-speed camera and fixed stage
    • live cell imaging (Calcium imaging and voltage-sensitive dye imaging)
  • Stereoscope with fluorescence and video capture
    • used for behavioral observations and optical imaging
  • Inverted microscope for anatomical studies
  • Intracellular and extracellular recording amplifiers
    • measures neuronal activities
  • Voltage clamp, current clamp, patch clamp and dynamic clamp systems
    • measures ionic currents in cells
  • In vivo electrophysiology system
    • for studying neuronal activity in living animals
  • Electrophoresis rig for gels
  • Microinjection/ picospritzer
    • for injecting DNA/RNA and dyes into cells or tissue
  • Laser-photoablation to selectively disable neurons
  • Thermocycler for amplifying DNA
  • QM-4 fluorimeter
  • Variety of high speed & ultra-centrifuges, micro centrifuges
  • Triaxial magnetic coil system for magnetic field manipulation in behavioral experiments
  • Unix and PC clusters for computer modeling
  • A 32 channel EEG/ERP system for monitoring brain activity
  • An eye-tracker for monitoring eye movements and pupil size
  • Voltammetry rig to measure in vivo changes of neurotransmitter release
  • Implantable voltammetry device and stimulator
    • for controlling neurotransmitter release from dopaminergic neurons

Teaching collections


For use in teaching we have a diversity of living and preserved organisms

Felmley Greenhouses

These greenhouses have a wide diversity of plants with a focus on tropic and economically important plants

George S. Vasey ISU Herbarium

Collection of over 50,000 specimens of plants dating back to 1859. For more information see Dr. Armstrong’s website who is the curator of this collection

John Wesley Powel and Dale Birkenholz Natural History Collections

Collections from 1867 of vertebrates including: fish, herps, birds and mamals. For more information see Dr. Capparella’s website who is the curator of this collection.