Laboratory for functional Fluorescence Microscopy Imaging (fFMI)

Laboratory for functional Fluorescence Microscopy Imaging (fFMI)

 

 

The Laboratory for functional Fluorescence Microscopy Imaging (fFMI) offers training and efficient access to standard and advanced fluorescence microscopy imaging instrumentation for CMM and external users at all levels of experience. Our objective is to support research on molecular and cellular mechanisms underlying normal physiological and pathophysiological processes. To this aim, we enable fluorescence microscopy imaging of fixed and live cells, and quantitative biochemical studies in live cells using Fluorescence Correlation Spectroscopy (FCS) and Fluorescence Cross-Correlation Spectroscopy (FCCS). These techniques enable quantitative characterization of molecular interactions in live cells with high spatial (200 nm) and temporal (µs) resolution and single-molecule sensitivity.

Four dedicated systems are presently available in the Laboratory for Functional Fluorescence Microscopy Imaging (fFMI).  This core facility is situated in the Center for Molecular Medicine (CMM), L5:02, rooms 020a (Systems 1-3) and 025 (System 4).

 

1. The ConfoCor 3 System

 

The ConfoCor 3 System, Carl Zeiss, Jena, Germany, is individually modified to allow fluorescence imaging with single-molecule sensitivity in live cells. This LSM 510 system consists of an inverted microscope for transmitted light and epifluorescence (Axiovert 200 M) with 6 objectives (5X Plan-Apochromat/0.16, 10X Plan-Neo/0.3, 20X Plan-Apo/0.75 with DIC capability, C-Apochromat 40X/1.2 NA W Corr, C-Apochromat 63x/1.2 W Corr, Plan-Apochromat 100x/1.40 Oil DIC), 3 lasers with 6 laser lines (Ar/ArKr laser with four lines 458, 477, 488 and 514 nm, HeNe 543 nm and HeNe 633 nm), HBO and Halogen lamps, 3 photomultiplier tubes (PMTs), scanning module LSM 510META modified to enable detection in the imaging mode by using silicone avalanche photodiodes (SPCM-AQR-1X; PerkinElmer) and FCS module with 3 detection channels.

 

Responsible:                     Vlad Radoi, PhD student; E-mail: vlad.radoi@ki.se

Booking ConfoCor3:        Booking system for instruments

 

 

2. ConfoCor2 System

 

The ConfoCor 2 System, Carl Zeiss, Jena, Germany for quantitative in vitro studies of molecular interactions. The ConfoCor2 system consists of an upright microscope with the C-Apochromat 40X/1.2 NA W Corr UV-VIS-IR objective, 3 lasers (Ar laser with three lines 458, 488 and 514 nm, HeNe 543 nm and HeNe 633 nm), a Perkin Elmer APD detection system and computer working station.

 

Responsible:                     Dr. Ann Tiiman, Associate Professor; E-mail: ann.tiiman@ki.se

Booking ConfoCor2:        Booking system for instruments

 

 

3. mpFCS-alpha System

 

The mpFCS-alpha System is a state-of-the-art home built microscope specially designed for quantitative, scanning free ultra-fast confocal fluorescence microscopy imaging of fast dynamical processes in live cells. To this aim, the system relies on massively parallel Fluorescence Correlation Spectroscopy (FCS).

 

Responsible:                      Dr. Sho Oasa, Postdoctoral Research Fellow; E-mail: sho.oasa@ki.se

Booking mpFCS-alpha:     Booking system for instruments

 

 

4. The mpFCS-beta System

 

The mpFCS-beta System is a state-of-the-art home built microscope specially designed for quantitative, scanning-free, ultra-fast confocal fluorescence microscopy imaging of fast dynamical processes in live cells by massively parallel Fluorescence Correlation Spectroscopy (FCS) and Fluorescence Lifetime Imaging Microscopy (FLIM).

 

Responsible:                     Dr. Sho Oasa, Postdoctoral Research Fellow; E-mail: sho.oasa@ki.se

Booking mpFCS-beta:     Booking system for instruments

 

5. To come: Funding for a new Confocal Laser Scanning Microscope (CLSM) system is approved by the CMM Steering Board. Installation of the new instrument is expected in late autumn 2018. 

Auxilary equipment: Microscope stage cell incubator consisting of heating Insert P, a solid heating element with uniform heat distribution and a high thermal conductivity, the CTI-Controller 3700 digital, with an IR absorption based CO2-sensor for continuous monitoring of CO2 concentration in the circulating air stream, and the Tempcontrol 37-2 digital, temperature regulator with two independent channels designed to maintain cells at the desired temperature and under desired atmosphere on a microscope stage.

 

Head of the Laboratory for functional Fluorescence Microscopy Imaging (fFMI)

Vladana Vukojevic, Associate Professor
Department of Clinical Neuroscience
Karolinska Institute

Contact number
Phone: 08 517 717 22
Mobile: 070 306 06 48

E-mail: vladana.vukojevic@ki.se

Location

CMM L5:02 Rooms 020a (systems 1-3) and 025 (system 4).

Training

Training is required for all first-time users at the cost of 1000 SEK. Training is limited to instructions on how to operate the system and use the software in order to acquire images, perform FCS measurements and export the results. Training does not include supervision in assay setup, assay preparation or result analysis. To schedule training sessions, contact us directly on the e-mail addresses above.

Costs

Supervised use: 350 SEK/h for CMM users and 450 SEK/h for users outside of CMM.
Unsupervised use during regular working hours (9-18 h): 150 SEK/h for CMM users and 250 SEK/h for users outside of CMM.
Unsupervised use outside of working hours (during the weekend and/or from 18-9 h): 50 SEK/h for CMM users and 150 SEK/h for users outside of CMM.
Service costs, including CMM IT costs, will be charged in accordance with the actual annual use.
Reagents have to be purchased by the user.

 

PhD course on Functional Fluorescence Microscopy Imaging (fFMI) in Biomedical Research
(next occasion HT18, Nov 19 – 30):

Functional Fluorescence Microscopy Imaging (fFMI) in biomedical research  

Doctoral education Aii - Allergy, immunology and inflammation