FV1000 (Olympus)

1) Application of the FluoView™ FV1000

• spectral multi-color imaging (single optical sections or 3D stacks) - spectral detection system allows very high sensitivity and combination of up to 4 colors of fluorescent markers time laps imaging (single optical sections or 3D stacks)
• multiposition time laps
• spectral deconvolution
• bleaching/photo-activation experiments

2) Instrument specifications

Confocal microscope Olympus FluoView™ FV1000 is an inverted IX81 motorized microscope equipped with:

Four lasers:

• UV laser diode (405 nm, 50 mW)
• Argon laser, multi-line: 457/488/515 nm (40 mW)
• yellow diode laser (559 nm, 20 mW)
• red diode laser (635 nm, 20 mW)

The selection of the laser line is controlled via an Acousto Optical Tunable Filter (AOTF). The use of an AOTF allows excitation intensity balancing within an image, flexible area bleaching e.g. for FRAP/FLIP/Photo-activation experiments and line sequential scanning to avoid cross-talk between channels (multi-tracking).

 

Excitation dichroic mirrors:

• DM 405/488/559/635
• BS 20/80
• DM 405/488
• DM 458/515

 

Emission dichroic mirrors and barrier filters:

Channels	Dichroic mirror (nm)	Barrier Filters (nm)
1 (PMT)	SDM490, SDM510, SDM560, SDM640, glass, mirror	Variable Barrier Filter
2 (PMT)	SDM560, glass, mirror	Variable Barrier Filter
3 (PMT)	glass, mirror, SDM640	BA567-587, BA575-620, BA575-675
4	-	BA655-755 (fixed)

 

Five detectors:

• 3 photomultiplier tubes (PMTs) for normal imaging. Two of the PMTs are equipped with a galvano grating spectral scanning systems for tunable bandwidth imaging which allows a fluorescence emission detection with variable setting of wavelength band and spectral imaging. This allows fine tuning of the emission detection and high speed spectral scanning. The spectral range of the PMTs detectors is appr. 300 nm to 850 nm
• 1 transmitted light detector for brightfield or DIC imaging. This setup allows for three fluorescent dyes to be imaged simultaneously along with another channel for transmitted light
• 1 PMT for the simultaneous scanner (SIM). SIM allows simultaneous stimulation and detection of signal in a variety of applications such as FRAP (Fluorescence Recovery After Photobleaching), FLIP (Fluorescence Lifetime Imaging Microscopy) photo activation, photo conversion, uncaging, etc. The system is equipped with the program for FRET (Fluorescence Resonance Energy Transfer) analysis.

 

Objectives:

The Olympus IX-81 inverted microscope can hold a total of 6 objectives at the same time. We currently have installed objectives listed in the table. In case you needed another one from all available Olympus objectives list, please, contact Hana Sehadova.
Description and magnification	N.A.	W.D. (mm)	Cover glass thickness	Immersion	Correcting rings	Condenser	U-DICTS position
UPLSAPO 10x	0.40	3.10	0.17	-	-	DIC10	normal
LUCPLFLN 20x	0.45	6.4-7.8	0-2	-	Yes	DIC20	normal
UPLSAPO 20xO	0.85	0.17	-	Oil	-	DIC20	normal
UAPON 40xO	1.35	0.10	0.17	Oil	-	DIC40	BFP1
PLAPON 60xOTIRFM	1.45	0.10	0.13-0.19	Oil	23°C, 37°C	DIC60	BFP1
UPLSAPO 100xO	1.40	0.12	0.17	Oil	-	DIC100	normal

 

Software:

• FLUOVIEW
• FLUOVIEW 3DViewer

3) FluoView™ FV1000 booklets and user manuals

Olympus flyers:

• Olympus FV1000 Confocal brochure 1
• Olympus FV1000 Confocal brochure 2
• Olympus FV1000 Confocal brochure 3
• Olympus IX81 Microscope brochure

 

Olympus user manuals

• Fluoview FV1000 quick start manual
• Fluoview FV1000 user manual
• Fluoview FV1000 safety guide
• Fluoview FV1000 hardware manual
• IX81 hardware

4) Theory of confocal microscopy

• Olympus website
• Nicon website
• Laser Scanning Confocal Microscopy - Nathan S. Claxton, Thomas J. Fellers, and Michael W. Davidson
• Presentation - Hana Sehadova
• Mikroskopy v rukou biologů - článek v časopise Vesmír