In situ/imaging facility

In situ/imaging facility

The former core facilities at CMM for mRNA in situ hybridization and phosphor imaging have merged into a new core facility for in situ hybridization and imaging. We are located at the Neurogenetics Unit, CMM L8:00, Karolinska University Hospital Solna.
Our function is to provide mRNA in situ hybridization and some related imaging methods to users at CMM and elsewhere.

WE OFFER
• In situ hybridization to measure and locate specific messenger RNAs in tissue. We have extensive experience with brain tissue, but have also worked with other tissues such as adipose tissue and heart.
• Light microscopy for in situ hybridization, conventional histology, immunohistochemistry etc.
• Documentation and quantification of experiments involving films, blots and gels.

FLEXIBLE SERVICES – TWO SERVICE LEVELS
Full-service:
The user provides us with tissue samples and we are performing the analysis.
”Hands-on” service:
The experienced user receives a short introduction by our personnel regarding methodology and operation of the instruments, then reserves a date and time for the use of an instrument.

ABOUT mRNA IN SITU HYBRIDIZATION
The detection of gene expression or viral sequences at the single cell level makes it possible to determine the site of production of molecules or pathogens in many different tissues. The use of high-resolution quantitative evaluation makes it possible to study gene regulation at single cell level as well as to evaluate effects of physiological or pharmacological manipulations. The facility enables guests or research groups to perform in situ hybridization on their specimens in a controlled environment to minimize set up time and rapidly produce results. The facility aims particulary at projects involving human genes, disease genes and human organs.

INSTRUMENTS AND METHODS AT THE FACILITY
A list of our equipment can be found below. To accommodate users with a need for assistance as well as experienced users, the core facility offers both full service assignments as well as the possibility for experienced users to access the equipment themselves.

FUJIFILM FLA-5100
• The FLA-5100 imaging system allows imaging of radioisotopic and fluorescent samples as large as 40 x 46cm at pixel size as low as 10 microns. This system is a rapid alternative to film in autoradiography experiments involving tissue sections or blots. It can also be used with protein electrophoresis in proteomics and in general array analysis for genomics.
• Currently equipped with three lasers: 473 nm for SYPRO Orange, SYPRO Ruby, SYBR Green I, FITC. 532 nm for Cy3, rhodamine and SYPRO Red. 635 nm for Cy5.
• Dynamic range five orders of magnitude.
• IP, fluorescence, digitizing function and chemiluminescence detection ability allows multipurpose use.
• Maximum sample size 460 x 400 mm.

ZEISS AXIOPHOT
• Light microscope equipped for bright field (both transmitted and reflected), phase contrast, transmitted and reflected polarization and Nomarski differential interference contrast microscopy, as well as epifluorescence. The microscope is equipped with color CCD camera.

THERMO SCIENTIFIC CRYOSTAR NX70
• Knife / Blade Holder cooling.
• Specimen cooling.
• Ergonomic height adjustment with integrated ergonomic casing.
• Colour touch-screen user interface and joystick control.
• Motorized cutting with variable speed control (0-256 mm/s).
• Automatic 24 hour defrost cycle.
• Display counter for every section.
• Section thickness range 0.5-500 um.

 

CONTACT INFORMATION
Ida Nilsson, PhD
Location: CMM floor 00
Phone: +46 (0)8- 517 764 02
E-mail: Ida.Nilsson@ki.se
Professor Martin Schalling
Location: CMM floor 00
Phone: +46 (0)8- 517 726 63
E-mail: Martin.Schalling@ki.se