Confocal Microscopy
Céline Hoffmann
Address: 84 Val Fleuri - 1526 Luxembourg
Phone. (+352) 26 970 253
Fax. (+352) 26 970 390
celine.hoffmann@crp-sante.lu
Confocal Microscopy
The platform was created in 2005 and established in the BAM building of the CRP-Santé. It is dedicated to cell biology research and confocal microscopy imaging.
The facility provides assistance to CRP researchers at different levels, such as sample preparation, microscope operation and image analysis. The platform is also open to academic or industrial users from Luxembourg and surrounding areas.
We have expertise in brightfield microscopy (DIC and phase contrast), epi-fluorescence and confocal microscopy. Currently, we are running several types of experiments, such as immuno-labelling, GFP observation and time-lapse series. Several types of software are available to process pictures in 3D or to extract quantitative data from your images.
-----------------------------------------------------------------------------
Photo contest: "Picture of the month", December
In vitro actin filaments polymerisation visualised
by TIRF Microscopy
Hoffmann Celine, CM
Confocal microscopy allows imaging of cells or tissues; on fixed samples or live cells in 3 dimensions; in one to 4 colors. My main activities on the platform are to ensure training of new users and to offer technical support to prepare samples or to set up and use the confocal microscope. Some examples from current experiments and results are presented below.
- Immuno-labeling and GFP imaging
- 3D reconstructions and animations
- Time-lapse experiments
- Quantification and counting
- Cell physiological state observations
-------------------------------------------------------------------------------------------------
| "Picture of the month", October 
F-Actin dynamics in presence of Actin Depolymerization Factor Visualized by TIRF microscopy Hoffmann Celine, CM |
|
| ----------------------------------------------------------------------------------- |
|
-------------------------------------------------------------------------------------------------------------------------- |
| "Picture of the month", May FEBS Letters Cover, Tholl et al., LBMV 
ADF9 (Actin depolymerisation Factor 9) |
"Picture of the month", June 
Actin (red) and vinculin (green) labelling on Vero cells , Moreau Flora, LBMV |
|
| ------------------------------------------------------------------------ |
||
| "Picture of the month": March 
"Art & Cells" Siti Aminah Abdul Rahim, LRNO |
"Picture of the month": April 
"Chondrocyte differentiation from human mesenchymal stem cells in alginate beads", Etienne Moussay and Jérôme Paggetti, LEHO |
|
| ------------------------------------------------------------ |
------------------------------------------------------------ |
|
| "Picture of the month": January 
Actin cytoskeleton labelling in U87 cells Phalloidin-Alexa488 dye and DAPI staining) |
"Picture of the month": February 
Phagocytosis of fluorescent particles by Human bone marrow mesenchymal stem cells, Etienne Moussay and Jérôme Paggetti, LHCE |
|
| ---------------------------------------------------------------- |
- |
------------------------------------------------- |
| 2010 Plant Cell cover: Papuga et al., LBMV 
Arabidopsis LIM Proteins: A Family of Actin Bundlers with Distinct Expression Patterns and Modes of Regulation. (2010) 22 : 3034-3052. |
||
| -------------------------------------------------- |
- |
-------------------------------------------------------------------------- |
| 2009
Tobacco BY2 Nucleus (blue) and 
|
2008 Tobacco parenchyma cells, LBMV 
|
|
| -------------------------------------------------- |
- |
-------------------------------------------------------------------------- |
| 2007
Blood vessel in tumor expressing 
|
2006
Tobacco leaf epidermal cells 
|
|
Equipment
Microscope : Axiovert 200M (Zeiss)
Confocal system : LSM510 META
TIRF system: Zeiss
Software : Imaris, Huygens, Metamorph
General information:
- Samples can be observed on microscope slides as well as in chambers (25 or 42 mm diameter), in Petri dishes or multiwell plates, but you have to use glass bottom dishes for high magnification or Long Distance objectives for plastic dishes.
- Remember to fill the logbook on the intranet, the application is called microscopy: date, name, time etc, when you start and when you stop.
- Immersion oil can damage the inverted objectives, always clean the objectives after use and clean immediately if you accidentally immerse the water objective (cleaning solution: ether petroleum 95%, isopropyl-alcohol 5%).
-
When you stop:
If the next user comes in less than one hour, leave the system on, put the laser argon on standby; but make sure the next person will come. - If the next user has booked for later than in one hour, shut down the system.
- Please ask if you have any doubt how to operate the microscope.
