Overview

Light is one of the most powerful tools to probe biology. We make it available to research laboratories at Stowers Institute. Our focus is on high resolution light microscopy, confocal microscopy (LSM) , non-linear optics (NLO) and fluorescence correlation spectroscopy (FCS) . We work in close collaboration with the Image Center.

Stowers Links:
Research Faculty
Image Center
Adv. Instr. & Physics: Microscopy
Adv. Instr. & Physics: Technology
Adv. Instr. & Physics: Two-Photon Microscopy
Adv. Instr. & Physics: FCS
Image Center internal

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All technologies we are currently using are based on light microscopy. To get three-dimensional information we use confocal Laser Scanning Microscopes (LSM). To image thicker, highly scattering and sensitive live samples we have combined some of these systems with two photon excitation. Fluorescence Correlation Spectroscopy (FCS) allows us to probe molecular diffusion and interaction in solution and living cells.

In most cases we rely on fluorescence as contrast mechanism, but we are working on Fluorescence Lifetime Imaging (FLIM) and Second Harmonic Generation (SHG) as well.

For data evaluation we use standard software packages. For special applications we use IDL and Mathematica.

For details, please visit our Technology pages:

• Microscopy - Summary at Technology
• Microscopy - In-Depth-Page
• Building Blocks of a Light Microscope
• From Sample to Eye
• Beam Path of Microscope with Infinity Optics
• Objective and Tube Lens
• Eyepiece in Microscope Beam Path
• Transmitted Light Illumination
• Resolution and Magnification
• Calculate Microscope Resolution Using Excel Spreadsheet
• Köhler Illumination
• Objectives
• Fluorescence
• Confocal Laser Scanning Microscopy (LSM)
• Two-Photon Microscopy, Non-Linear Optics (NLO) - Summary at Technology
• Two-Photon Microscopy, Non-Linear Optics (NLO) - In-Depth Page
• Two-Photon Microscopy
• Two-Photon Excited Fluorescence
• Absorption of Two Photons
• Excitation Spectra
• Absorption of NIR Light in Tissue
• Instrumentation (Carl Zeiss LSM 510 META)
• Pulsed Near-Infrared Laser (Coherent Chameleon)
• Fluorescence Lifetime Imaging Microscopy (FLIM)
• Fluorescence Correlation Spectroscopy (FCS) - Summary at Technology
• Fluorescence Correlation Spectroscopy (FCS) - In-Depth Page
• Molecular Diffusion and Interaction
• The Principle: Exploiting Fluctuations
• Source of Fluctuations
• Create a Small Measurement Volume
• Detect Single Photons
• Calculate Correlation Function
• Literature FCS
• Software - Summary at Technology
• Software - In-Depth Page
• Carl Zeiss Confocal Microscope (AIM) Software
• AIM Macros
• Software for Fluorescence Correlation Spectroscopy (FCS)
• Mathematica
• ImageJ
• IDL

Stowers Links:
Adv. Instr. & Physics: Technology
Adv. Instr. & Physics: Microscopy
Adv. Instr. t& Physics: Two-Photon Microscopy, Non-Linear Optics (NLO)
Adv. Instr. & Physics: FCS
Adv. Instr. & Physics: Software
Image Center: Instruments

All of our projects are in close collaboration with laboratories at the Stowers Institute. This page describes some of the topics we are working on, including transcription involving RNA-Polymerase II, imaging of the fly brain and Second Harmonic Generation (SHG) to measure membrane potentials. These pages are mainly intended for internal communication, but feel free to browse them.

For details, please visit our Projects page:

• Microscopy: Imaging the Mouse Eye
• Two Photon Microscopy: Imaging the Fly Brain
• Fluorescence Correlation Spectroscopy: Interaction between RNA-Polymerase II and Co-Factors

Stowers Links:
Adv. Instr. & Physics: Projects
Research Faculty

Technology & Methods

On the last Thursday of every month, we organize a Technology and Methods Seminar. We present technologies offered at the Stowers Institute and their applications.

Foundations of Microscopy

On the 1^st and 3^rd Friday of the month we discuss fundamentals in microscopy and the related theoretical basics. These seminars are intended for people who want to understand the inner workings of a microscope in order to improve their scientific results based on optical imaging or modify and built microscopes and related optical set-ups.

Foundations of Image Processing

Image processing is becoming a major part of microscopy. On the 4^th Friday of the month we discuss theory and implementation of image processing algorithms.The seminars are intended for users of image processing software who want to ensure optimal results while avoiding artifacts. We will discuss different software implementations and how to develop new algorithms.

FCS User Club

Once a month we are disussing classic papers about FCS and related techniques. The FCS User Club is a forum for current and potential users of fluorescence correlation spectroscopy.

For details, please visit our Seminars page:

• Technology & Methods: Future Presentations
• Technology & Methods: Presentations 2006
• Technology & Methods: Presentations 2005
• Foundations of Microscopy
• Foundations of Image Processing
• FCS User Club

Stowers Links:
Adv. Instr. & Physics: Seminars
Stowers Bioplog

Director of Advanced Instrumentation & Physics is Winfried Wiegraebe. Joseph Huff focuses on Fluorescence Correlation Spectroscopy. Amanda Combs does Two-Photon- and Second-Harmonic-Imaging. We are supported by our administrative assistant Whitney Bartlow.

For details, please visit our People page:

• Winfried Wiegraebe
• Amanda Combs
• Christopher J. Wood
• Joseph Huff
• Laura Laszczynski

Stowers Links:
Adv. Instr. & Physics: People

We have collected links to references, companies and downloads related to microscopy and the projects we are working at. In addition you will find some information about Kansas City.

Please let us know, if you come across some web pages we should add.

For details, please visit our Links page:

• References
• General
• Microscopy
• Dyes
• Biology
• Chemistry
• Literature
• Language
• Conferences and Workshops
• Companies
• Microscopy and Optics
• Microscopy
• Optical Filters
• Illumination
• Optical Components
• Fiber Optics
• Accessories
• Coverslips etc.
• Fluorescence Lifetime Systems (FLIM)
• Environmental Control
• Detectors
• Lasers
• Dyes and Slide Preparation
• Biology and Biochemistry
• Tools
• Downloads
• Presentations
• Literature
• Microscopy
• Confocal Microscopy
• Fluorescence
• Optical Filters
• Fluorescence Lifetime Imaging Microscopy (FLIM)
• Fluorescence Correlation Spectroscopy (FCS)
• Detectors
• Software
• Manuals
• Stowers Institute
• Public Pages
• For Internal Use Only
• Kansas City
• Arts
• Music
• General

Stowers Links:
Adv. Instr. & Physics: Links

Stowers Links:
Adv. Instr. & Physics: Site Map