Seitenhistorie

Location: Altensteinstr. 23a (SupraFAB), lab 026.1 (OM S2 area)

SN: 8100000277

FUB: 7200001673

Detailed information and protocols how to use OpenIris you find here.

Requirements:

e.g. device specific training & safety briefing:

1. Watch the OM video tutorial

1. and the video about confocal microscopy.
2. Register as new user for SupraFAB by writing to safety@suprafab.fu-berlin.de, provide personal info (name, working group, status/activity, office and laboratory room number) and confirm your particiaption in the next SupraFAB safety briefing taking place usually the first Monday of the month, 10:00 - 12:00 (if you do not get a confirmation, please write another email or contact the SupraFAB coordinator or secratary in room 102 or 203, see SupraFAB Homepage). 

Requirements e.g. device specific training & safety briefing:

1. Watch the OM video tutorial.
2. Participate in person in the annual SupraFAB safety briefing (next: June 3rd 2024 at10am, room 119);
   in case you need access before, watch the recorded safety briefing & become familiar with the safety guidelins, instructions and rules & biolab agreement.
3. Fill the transponder form with work safety questionaire(2. page), let it sign by your research group head and the respective responsible persons in supraFAB and finally hand in to Katharina Tebel (office 103) or Achim Wiedekind (office 102). 

4. Become familiar with the safety guidelins, instructions and rules.

5. Make an appointment with your Keyuser*, to get practical training (2-3 session) & an intro to the OM, the BioSupraMol, data handling (network) as well as the SupraFAB House guidelines. .
   In case you are the first person of your group who wants to use the device or in case of fuurtehher questions, contact the responsible person shown in OpenIris (https://fub.openiris.io/timeline/4c597b95-835a-4b7d-b69c-00f7f43df1c6).
6. Follow the addtional info for full accesss on the Access to the Optical Microscopy (OM) Facility page.
7. Book the device using OpenIRIS (protocls see here).

Note: Scientific guests as well as non-scientific staff such as service personal from companies also need to get introduced by one of the PIs for genetic engeeneering (see Biolab Team & Lab Phone Numbers). The introducution and documentation forms and slides can be found on the safety intro page (https://wikis.fu-berlin.de/x/pQhVT). The documentation forms and permission forms need to be stored in the repsective folder in office 111

Confocal Laser Scanning Microscope Leica SP8 (1)

Confocal laser scanning microscopy enables optical sectioning of multilayer fluorescent specimens with high contrast by applying a pinhole in the optics. It is therefore possible to image a thin optical slice out of a thick specimen (up to 100 µm) that represents under optimal conditions a slice of approximately 500 nm. Moreover, conventional contrast methods such as the differential interference contrast (DIC) can be used and in reflection mode particles or surfaces can be analyzed.

The CLSM SP8 (1) based on a DMI6000CSB allows precise detection of standard fluorophores in your sample. The system excites via diode laser at 405 nm, Argonlaser combined with AOBS at 458, 488, 514 nm, diode laser at 561 nm and a He/Ne laser at 633 nm. The SP8 (1) has two standard GaAsp photomultipliers (PMTs) and two extremely sensitive Hybrid detectors (HyD), combining PMT and avalanche photodiode (APD) technology. An optional incubation chamber allows imaging of living cells.

The system is almost fully automated and offers with its broad excitation spectra, its automated system parts, the galvo stage and the optional incubation chamber a variety of possible uses.

Immersion objectives are optimized for oil- / glycerol immersion.

Detailed specifications CLSM Leica SP8 (1)

Microscope:

• Leica DMI6000CSB stand (inverted)
• Motorized scanning stage
• Heated incubation chamber with CO supply from LIFE IMAGING SERVICES

Lasers:

• 405 nm Diode laser
• Argon/2 (458, 488, 514nm)
• 561 nm Diode laser
• HeNe 633nm

Conventional fluorescence filters for eyepiece vizualization:

• Blue (A = xBP340-380, dichroic 400, LP425)
• Green (I3 = xBP450-490, dichroic 510, mLP515)
• Red (N2.1 = xBP515-560, dichroic 580, mLP590)

Condenser:

• Smart condenser S28/0.55 w/o flip

Objectives:

• 5x/0.15 HCX PL FLUOTAR WD 13.7 mm
• 20x/0.75 HC PL APO Imm Corr (oil, water, glycerol) WD 0.68 mm
• 40x/1.30 HC PL APO Oil CS2 WD 0.24mm
• 63x/1.4 HC PL APO CS2 OIL WD 0.14 mm
• 63x/1.3 HC PL APO CS2 Glyc WD 0.30 mm

Dichroic(s):

• AOBS

Detectors:

• All spectral detectors: four channels - two PMTs, two HyDs - high sensitivity Hybrid Detectors - HyD1 | PMT2 | HyD3 | PMT4
• Motorized transmitted light detector

Computer:

• HP Z420 Workstation, Windows 7-64 bit, LAS X

Konfokales Laser-Scanning-Mikroskop Leica SP8 (1)

Die konfokale Laser-Scanning-Mikroskopie ermöglicht es durch Einsatz einer Lochblende, optische Schnittbilder mit hohem Kontrast von mehrschichtigen fluoreszierenden Präparaten zu erzeugen. Daher ist es möglich, aus einer dicken Probe (bis ca. 100 μm) eine dünne Präparatschicht abzubilden, die unter Idealbedingungen einer Schichtdicke von weniger als 500 nm entspricht. Darüber hinaus können auch herkömmliche Kontrastverfahren wie z.B. der differentielle Interferenzkontrast (DIC) genutzt werden und im Reflexionsmodus Partikel oder Oberflächen analysiert werden.

Das CLSM SP8 (1) auf Basis des DMI6000CSB erlaubt eine punktgenaue Detektion von Standard-Fluorophoren in unterschiedlichen Medien. Die Anregung der Fluorophore erfolgt über einen Diodenlaser bei 405 nm, einen Argonlaser mittels AOBS Strahlteiler bei 458, 488, 514 nm, einem Diodenlaser bei 561 nm und einem He/Ne Laser bei 633 nm und erlaubt somit die Verwendung der gängigen Fluroreszenzfarbstoffe. Die Detektion erfolgt über zwei konventionelle Photomultiplier (PMTs) bzw. zwei extrem sensitive Hybriddetektoren (HyD), welche Elemente eines PMT mit einer Avalanche-Photodiode kombinieren. Durch Verwendung einer optionalen Inkubationskammer können mit dem System lebende Zellen untersucht werden.

Das System ist nahezu vollständig automatisiert und erlaubt durch sein breites Anregungsspektrum und die vielseitigen automatisch getriebenen Systemeinstellungen, der Galvo-Stage und der optional einsetzbaren Inkubationskammer vielfältige Nutzungsmöglichkeiten. Die Immersionsobjektive sind für Öl- bzw. Glycerin-Immersion ausgelegt.

Detaillierte Spezifikationen CLSM Leica SP8 (1)

Microscope:

• Leica DMI6000CSB stand (inverted)
• Motorized scanning stage
• Heated incubation chamber with CO supply from LIFE IMAGING SERVICES

Lasers:

• 405 nm Diode laser
• Argon/2 (458, 488, 514nm)
• 561 nm Diode laser
• HeNe 633nm

Conventional fluorescence filters for eyepiece vizualization:

• Blue (A = xBP340-380, dichroic 400, LP425)
• Green (I3 = xBP450-490, dichroic 510, mLP515)
• Red (N2.1 = xBP515-560, dichroic 580, mLP590)

Condenser:

• Smart condenser S28/0.55 w/o flip

Objectives:

• 5x/0.15 HCX PL FLUOTAR WD 13.7 mm
• 20x/0.75 HC PL APO Imm Corr (oil, water, glycerol) WD 0.68 mm
• 40x/1.30 HC PL APO Oil CS2 WD 0.24mm
• 63x/1.4 HC PL APO CS2 OIL WD 0.14 mm
• 63x/1.3 HC PL APO CS2 Glyc WD 0.30 mm

Dichroic(s):

• AOBS

Detectors:

• All spectral detectors: four channels - two PMTs, two HyDs - high sensitivity Hybrid Detectors - HyD1 | PMT2 | HyD3 | PMT4
• Motorized transmitted light detector

Computer:

• HP Z420 Workstation, Windows 7-64 bit, LAS X