Functional Fluorescence Microscopy Imaging (fFMI)
The Laboratory for functional Fluorescence Microscopy Imaging (fFMI), Center for Molecular Medicine (CMM), 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.
Five dedicated systems are presently available in the Laboratory for Functional Fluorescence Microscopy Imaging (fFMI), L8:01, room 056.
Head of the Laboratory for functional Fluorescence Microscopy Imaging (fFMI):
Vladana Vukojevic, Associate Professor
Department of Clinical Neuroscience
Karolinska Institutet
Contact: E-mail: vladana.vukojevic@ki.se; Phone: 070 306 06 48
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. For new users, 5 supervised sessions are required, following the introductory training.
Unsupervised use during regular working hours (9-18 h): 100 SEK/h for CMM users and 200 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 100 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.
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; Phone: 073 647 81 04
First time users: If you are interested in using this system, please contact the responsible person for introductory training. System use without previous training is not allowed.
Booking ConfoCor3: Link to the booking system here.
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, Assistant Professor; E-mail: ann.tiiman@ki.se; Phone: 076 708 40 81
First time users: If you are interested in using this system, please contact the responsible person for introductory training. System use without previous training is not allowed.
Booking ConfoCor2: Link to the booking system here.
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
Phone: 072 287 09 53
First time users: If you are interested in using this system, please contact the responsible person for introductory training. System use without previous training is not allowed.
Booking mpFCS-alpha: Link to the booking system here.
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
Phone: 072 287 09 53
First time users: If you are interested in using this system, please contact the responsible person for introductory training. System use without previous training is not allowed.
Booking mpFCS-alpha: Link to the booking system here.
5. LSM 880 Confocal Microscope (without AiryScan)
The LSM 880 Confocal Microscope (without AiryScan), Carl Zeiss, Jena, Germany consists of an inverted microscope for transmitted light and epifluorescence (Axio Observer 7) with 4 objectives: 10X air (Plan-Apochromat 10x/0.45 WD=2.0 M27), 20X air (Plan-Apochromat 20x/0.8 WD=0.55 M27), 25X multi-immersion (LD LCI Plan-Apochromat 25x/0.8 Imm autocorr DIC M27 for water, silicone oil, glycerine or oil immersion, WD=0.57mm at CG=0.17 mm), and 63X oil (C Plan-Apochromat 63x/1.4 WD=0.17 Oil DIC M27), all allowing simultaneous acquisition of transmitted laser light and differential interference contrast (DIC) imaging; 4 lasers with 7 lines for excitation at 405 nm, 458 nm, 488 nm, 514 nm, 453 nm, 633 nm; Halogen and Mercury short-arc reflector lamp (Illuminator HXP 120) for transmitted and reflected light, respectively; 3 detection channels of which two are multialkali photomultiplier tubes (PMTs) and one GaAsP detector; Scan module LSM 880 with high-speed scanners. Dedicated for imaging fixed samples. Can be used for imaging live samples with the microscope stage cell incubator (see Auxiliary equipment). Software ZEN 2.3 SP1 FP3 (black). To transfer your images to the CMM file server, follow the instructions at: https://10.129.28.247/web/guest/service_catalogue/itservices/access-home-folder-windows#unmanaged
Responsible: Dr. Sho Oasa, Postdoctoral Research Fellow; E-mail: sho.oasa@ki.se
Phone: 072 287 09 53
First time users: If you are interested in using this system, please contact the responsible person for introductory training. System use without previous training is not allowed.
Booking LSM 880: Link to the booking system here.
Auxiliary 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.