Leica MacroFluo™ User manual
Dear User, Thank you for your faith in our product. We hope your work with it is both enjoyable and successful. Leica MacroFluo™ with zoom 6.3:1 or 16:1 are worldwide the only macro documentation systems for fluorescence. The MacroFluo™ concept combines the advantages of macroscopy – large fields of view, large working distances, parallax-free and precise imaging – with fluorescence technology (including multi color fluorescence) at a high resolution.
Overview of the instrument 13 12 11 9 10 8 7 5 2 14 15 6 4 1 3 Fig. 1 Leica MacroFluo™ with zoom system 16:1 1 Base of stand (transmitted-light base HL RC™ shown) 2 Focusing drive with column (shown with motor focus) and focusing stop (shown with holder for fiber-optic light guide) 3 Handheld controller (only if fitted with motor focus) 4 Gliding stage (optional), glass stage plate is standard 5 Zoom system 6.
Contents Page 1 Safety concept . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 1.1 The user manual . . . . . . . . . . . . . . . . . . . . . . . . . 5 1.1.1 Diagrams . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 1.1.2 Symbols used . . . . . . . . . . . . . . . . . . . . . . . 6 1.2 Safety instructions . . . . . . . . . . . . . . . . . . . . . . . . 6 1.3 Light source: Safety instructions . . . . . . . . . . . 9 2 Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1. Safety concept 1.1 The user manual Included with your Leica MacroFluo™ is an interactive CD-ROM with all pertinent user instructions in German, English, French, Spanish, Italian, Portuguese, Dutch, Danish, Swedish, Finnish, and Greek. Keep it in a safe place, and readily accessible to the user. User manuals and updates are also available for you to download and print from our web site at www.stereomicroscopy.com.
1.1.2 Symbols used 1.2 Safety instructions Description Warning of a danger This symbol indicates especially important information that, if not observed, – can cause hazards to personnel, – can lead to functional disturbances and damaged instruments. Warning of hazardous electrical voltage This symbol indicates especially important information that, if not observed, – can cause hazards to personnel, – can lead to functional disturbances and damaged instruments. Danger due to hot surface.
Permitted uses Place of use The Leica MacroFluo™ allows for examining, manipulating, sorting and documenting entire unprepared fluorescent objects. The high resolution, high light utilization and specially selected filter blocks allow for distinguishing the finest structures and increase the richness of information with incident light fluorescence. But the Leica MacroFluo™ can also be used to perform observations without fluorescence in transmitted light or incident light.
Repairs and servicing Legal requirements – Only Leica Microsystems-trained service technicians, or technical specialists assigned by the person in charge of the instrument, are permitted to carry out repairs. – Only original Leica Microsystems spare parts may be used. – Unplug the power cable, the power supply or the supply unit before opening voltage carrying parts such as the Leica MacroFluo™. Adhere to general and local regulations relating to accident prevention and environmental protection.
Direct contact with eyepieces is a potential transmission method for bacterial and viral infections of the eye. Users should be made aware of the potential risk of infection. The risk can be kept to a minimum by using personal eyepieces for each individual or detachable eyecups.
2. Assembly C-Mount 1x MacroFluo B-Mount 2/3” 2.1 Expansion diagram B-Mount 1/2” 11 543 706 11 543 702 C-Mount 11 543 703 11 541 512 0.35x C-Mount 11 541 511 0.5x Vario TV-Adapter, delta 11 541 038 0.55x - 1.1x 11 541 517 0.32x - 1.6x 0.55x - 1.1x 11 541 537 0.
Focusing drive ➜ Base of stand 2.
16 15 14 13 12 11 9 10 6 8 17 2 18 7 3 5 1 4 Fig.
2.5 Trinocular tube 2.6 Zoom system Trinocular tube ➜ incident illuminator The MacroFluo™ can be combined with a 6:1 (Z6 APO) or 16:1 (Z16 APO) zoom system. Tool: Allen key. 왘 Slightly unscrew the hexagon-head screw (3.9). 왘Insert the trinocular tube FSA HC L 3TP (3.8) in the circular mount (dovetail ring) and align by turning (view from the front). Ensure that the components do not jam. 왘 Tighten the clamping screw (3.9). Eyepieces – The designation on the eyepiece, e.g.
2.7 Objectives – We recommend using the 1×, 2×, 0.8×, 0.5× planapochromatic objectives, in order to take advantage of the high performance of the apochromatic zoom system. – Use the objectives directly on the diaphragm ring of the zoom system or with fine focusing. Remove the diaphragm ring if the fine focusing is used (see below). – A planapochromatic 5× HR objective is available for magnifications in the microrange (see assembly diagram p. 10). Always use the HR objective with fine focusing.
2.8 UV protection screen 왘 Fasten UV protection screen (5.5) with clamp (5.4) on the hinged arm (5.1). 왘 Fasten hinged arm with UV protection screen to the carrier using the clamp (5.2). 2 1 3 The hinged arm can be opened and closed via the central rotary knob (5.3). In the opened state, all joints are freely movable and the hinged arm can be placed in any position. To block all hinged arms, turn the rotary knob clockwise.
2.9 Lamp housing 2.10 Instrumentation of nosepiece disc The Leica lamp housing 106 z (11 504 069) is the light source for the incident illuminator LRF 4/22. It is used with 100-W mercury burners (11 500 138) or 103 W/2 (11 500 321) with extended life, the supply unit ebq 100 W and 103 W/2 (11 500 325) and straylight protection (10 446 271). The nosepiece disc can be fitted with a total of 4 filter blocks (filter system). At present, 19 fluorescence blocks (p.
The 4 positions of the nosepiece disc are each labeled with the numbers 1 through 4 on the left side of the dovetail guide. The numbers can be seen in the opened nosepiece slot. In addition, the active position in the beam path is displayed on the outside of the nosepiece disc (6.3). The filter blocks are supplied with self-adhesive labels numbered 1 through 4 and labels with short designations of the filter block (e.g. GFP).
2.11 Connection of digital or video cameras The trinocular tube is observation and photo tube in one and features a photo mount for adapting digital or video cameras. Various adapters as well as Vario TV adapters (with variable magnification) with C and B-mount are available. The image detail on the monitor depends on the adapter being used and the chip size of the camera. 1 Fig. 10 3 2 4 Photo mount 1 Hexagon-head screw for fastening a TV adapter in the photo mount 2.12.
3. Operation 3.1 Starting up 왘 Activate the lamp housing for the incident light fluorescence illuminator at the supply unit and adjust the burner. We recommend that you familiarize yourself with the operation of your Leica MacroFluo™ systems, such as adjusting the parfocality, zoom, focusing drive, eyebase, etc. in transmitted light and to experiment with it.
There are certain switchable zoom positions which can be activated for repetitive tasks: – For the Leica Z6 APO, these are 0.57 / 0.8 / 1 / 1.25 / 1.6 / 2 / 2.5 / 3.2 / 3.6. – For the Leica Z16 APO, these are 0.57 / 0.8 / 1 / 1.25 / 1.6 / 2 / 2.5 / 3.2 / 4 / 5 /6.3 / 8 / 9.2 The overall magnification is increased by the factor 1.25 of the trinocular tube HC L3TP (see the table on p. 30). 4 3 1 4 왘 Adjust the magnification by turning the knurled ring (12.1).
3.4 Iris diaphragm 3.5.1 Fine focusing at the objective The integrated iris diaphragm is used for continuous adjustment of the depth of field without changing the magnification. Reducing the diaphragm diameter enlarges the depth of field. The size of the light cone is also reduced, and the brightness of the image decreases. The optional fine focusing (12.4) allows for sensitive and precise focusing in a range of 10mm.
3.7 Adjusting diopter settings (parfocality) With adjustable eye lenses at the eyepieces, the observer can compensate his or her defective vision. The correct diopter compensation is required for both eyes so that focusing remains constant during zooming (parfocal). With a correct setting, you no longer have to refocus during zooming. If you turn the eye lens, an engraved silver line running all around on the outside of the eyepiece housing becomes visible.
1 2 6 8 7 5 Fig.
3.8 Incident light fluorescence illuminator 3.8.1 Field diaphragm The patented Leica zero-pixel-shift technology ensures an exact convergence for the superposition of different fluorescences. When the filter blocks are switched, no interfering pixel offset occurs between the individual images of the different spectral ranges – an ideal prerequisite for digital multicolor fluorescence experiments. The field diaphragm is used for reflection protection and specimen protection.
3.8.2 Aperture diaphragm 3.8.5 Filter blocks The aperture diaphragm is used for setting the contrast for reflecting specimens or, if necessary, for reducing the excitation radiation to prevent excessive bleaching of the specimens. The filter turret (14.1) can accommodate up to four filter blocks (ch. 2.10). These filter blocks contain the excitation filter, dichromatic beam-splitting mirror and suppression filter.
4. Appendix 4.1 Filter blocks for macro-fluorescence Filter block Fluorochrome Excitation Excitation filter Dichromatic Mirrors Suppression filter Order no.
4.2 Assignment of fluorochromes and filter blocks Fluorochromes Filter block – Acridine-blue A – Acridine-yellow I 3, H 3 – Acridine-orange I 3, H 3 – Acridine-red N 2.1, N 3 – Acriflavine E 4, H 3 – Acriflavine feulgen SITS (AFS) D – Alizarin complexion N 2.1 – Alizarin-red N 2.1 – Allophycocyanine (APC) Y 3, Y 5 – AMCA (Aminocoumarine) A – Aminoactinomycinn D-AAD N 2.1, N 3 – Aniline-blue A – ACMA E4 – Astrazon brilliant red 4G N 2.1 – Astrazon red 6B N 2.
– Calcein-blue A – Calcium crimson Y3 – Calcium green K 3, I 3, L 5 – Calcium orange M 2, N 2.
– FITC (FlouresceinIsoThioCyanate) I 3, H 3, K 3, L 5 – FITC/Ethidiumbromide I 3, L 5, N 2.1 – FITC (selective) L5 – Texas red (selective) TX 2 – FITC/TRITC L 5, N 3 – TRITC (selective) N3 – Fluo 3 I 3, L 5 – Fluoro Gold A – Fluram (Fluorescamine) A – Genacryl brilliant red B N 2.1 – Genacryl brilliant yellow E4 – Generic blue D – GFP (Green Fluorescent Protein) GFP – Granular blue A – Haematoporphyrin N 2.1 – Hoechst dye no. 33258 A, D, A 4 – No.
– Phycoerythrin (PE) N 2.1, N 3 – Primulin O D – Procion Yellow D, E 4, H 3 – Propidiumiodide N 2.1 – Pyronin B N 2.1, M 2 – Quinacrine mustard (QM) E4 – Resorufin N 2.1, Y 3 – Reverine D – Rhodamine B N 2.1 – Rhodamine 123 I 3, L 5 – Acid fuchsin N 2.1, M 2 – Serotine A, D – SITS (Stilben Iso Thio Sulfonic acid) A – SITS Acriflavine Feulgen D – Spectrum orange M 2, N 2.
4.3 Optical data Visual data Eyepieces HC Plan 10×/25 (11507808) Tube factor: 1.25× trinocular tube FSA HC L 3TP Iris diaphragm open Objectives Planapo 1× Planapo 2× Planapo 5× / 0.5 Planapo 0.5× Planapo 0.8× (10 447 176) (10 447 178) (10 447 243) (10 447 177) (10 446 360) 39mm 19.
4.4 Technical data, performance characteristics Designation Microscope type Zoom system Zoom factor Switchable zoom positions for repetitive tasks Tube Integrated iris diaphragm Fluorescence illuminator Type Fluorescence filter Light source Safety measures 32 Leica MacroFluo™ with 6.
Optical accessories Objectives Fine focusing Tubes Eyebase Large field eyepiece for spectacle wearers Type/magnification factor Working distance Planapo 1× 97mm Planapo 0.8× 112mm Planapo 2× 39mm Planapo 0.5× 187mm Achromat 0.63× 149mm Achromat 0.5× 187mm Achromat 0.32× 297mm Planapo 5×/0.5 LWD 19mm 10mm travel, 1 micron resolution – Trinocular tube FSA HC L 3TP (1.25×) – Trinocular tube HL L 2 TU 55mm – 75mm Wide-field HC plan eyepieces 10×/22, 10×/25 Max. field of view ø 35mm 44mm 17.
4.
∅58 155.6 118.5 ∅9.5 ∅9.5 ∅58 ∅58 37.2 ∅58 ∅63 50.6 ° 5.3° 44.6° 8.3° 37.
∅45 M42x3 12 54.8 38.2 9 6 ∅45 M42x3 12 ∅58 ∅45 M42x3 41.5 6 ∅58 ∅45 6 12 M42x3 32 ∅58 12 ∅45 M42 (Fine Focus Interface) ∅38 6.
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